PREPARATION OF N-ISOPROPYLACRYLAMIDE BASED

THERMALLY RESPONSIVE BLOCK COPOLYMERS FOR

CHEMICAL SWITCHING DEVICE

(N-イソプロピルアクリルアミドに基づく温度応答性ブロッ

ク共重合体の調製と化学スイッチングデバイスへの応用)

by

JIANMIN YANG

Dissertation submitted to the Department of Applied Chemistry

in Partial Fulfillment of the Requirements for the Degree of

Doctor of Engineering in Chemistry

at the

Tokyo Metropolitan University

Thesis Supervisor: Prof. Dr. Katsumi Uchiyama

September 2015
 PREPARATION OF N-ISOPROPYLACRYLAMIDE BASED
THERMALLY RESPONSIVE BLOCK COPOLYMERS FOR
CHEMICAL SWITCHING DEVICE
(N-イソプロピルアクリルアミドに基づく温度応答性ブロック共重合
体の調製と化学スイッチングデバイスへの応用)
by
JIANMIN YANG
Dissertation submitted to the Department of Applied Chemistry on Sep. 2015
in Partial Fulfillment of the Requirements for the Degree of
Doctor of Engineering in Chemistry

ABSTRACT

Poly(N-isopropylacrylamide) (PNIPAAm) is an excellent thermal responsive polymer

with a lower critical solution temperature (LCST) in water of 32 °C. The properties of

PNIPAAm can be altered by changing the temperature around LCST. When the

temperature is below the LCST, the intermolecular hydrogen bonds between the

PNIPAAm chains and the water molecules contribute to an extended brush structure

displaying hydrophilicity. When the temperature is above the LCST, the intramolecular

hydrogen bonds in the PNIPAAm chains could lead to a compact and collapsed

conformation with hydrophobic property.

This dissertation focuses on the preparation of NIPAAm based block copolymers and

their application in chemical switching device. The thermally responsive property and

applications of PNIPAAm are introduced in Chapter 1.

In Chapter 2, the copolymer, poly(N-isopropylacrylamide-co-hexafluoroisopropyl

acrylate) (poly(NIPAAm-co-HFIPA)), was initially synthesized via surface-initiated atom

transfer radical polymerization (ATRP) method. The HFIPA added to enhance the

i
hydrophobicity of the polymer. And the water contact angle (CA) was measured to

evaluate the surface characteristics. The results indicated that the copolymer showed

hydrophilic/hydrophobic transition with temperature across LCST. The copolymer has

great potential for use in various important applications, including water controllable

transportation, molecular separation, and so on.

In Chapter 3, a smart and reversible chemo-mechanical switch was developed for

controllable water transportation. The smart switch was constructed by integrating

poly(NIPAAm-co-HFIPA) on a capillary plate. With the temperature changing around

LCST, the designed chemo-mechanical switch exhibited excellent ‗‗ON–OFF‘‘ behavior

for water transportation. Meanwhile, the sequences of copolymer affected on properties of

switching have investigated. The excellent controllability over aqueous solution

transportation indicates that it has great potential for use in various important applications,

including intelligent microfluidic switching, water/oil separation, controllable drug release,

and so on.

In Chapter 4, the copolymer, poly(N-isopropylacrylamide-co-methacrylic acid)

(poly(NIPAAm-co-MAA)), was synthesized via free radical initiated precipitation

polymerization. To match the human physiological temperature, the MAA was added to

raise the LCST of the copolymer. The results showed the copolymer with the

NIPAAm:MAA molar ratio of 100:10 have a satisfactory LCST of 37.5 °C. Meanwhile, the

period for the copolymer conformation change was only 8 min. The synthesized copolymer
has great potential for use in human drug delivery system.

In Chapter 5, a straightforward approach for the preparation of monodisperse porous

polymer particles by inkjet technology was developed. The merit of the inkjet-based

method for polymer particles production was demonstrated by using sodium

poly(styrenesulfonate) (NaPSS) solution as a typical example. In which, NaPSS solution

can form monodisperse porous polymer particles in a single step. Subsequently, the
cross-linked 1,6-hexanediol diacrylate (HDDA) particles with hollow core-porous shell

ii
structure were produced by the combination of ink-jetting and UV polymerization

approach. The chemical and physical properties stability of HDDA particles have high

potential for used as drug carriers.

In Chapter 6, a thermo-responsive drug delivery system was fabricated by grafting the

copolymer poly(NIPAAm-co-MAA) onto the surface of HDDA particle. The drug

molecules can be enclosed when the temperature below 37 °C, where the pore channels

were closed by the swelling copolymer. Whereas as the temperature increasing, the

copolymers become collapsed, and the drug could released from the opening pore channels.

The drug controlled release behavior has been investigated.

In Chapter 7, the principal findings and results of this thesis are briefly summarized.

Keywords: N-Isopropylacrylamide; Thermally responsive polymers; Chemical

switching; Water controllable transportation; Drug controlled release.

iii
iv
 TABLE OF CONTENTS

ABSTRACT........................................................................................................... i

TABLE OF CONTENTS..................................................................................... v

LIST OF ABBREVIATIONS............................................................................... ix

CHAPTER 1: INTRODUCTION.................................................................. 1

1.1 Stimuli-responsive polymers......................................................................... 1

1.2 Thermally responsive poly(N-isopropylacrylamide)..................................... 3

1.2.1 Thermally responsive polymers............................................................ 3

1.2.2 Poly(N-isopropylacrylamide)............................................................... 5

1.2.3 Synthesis of poly(N-isopropylacrylamide)........................................... 7

1.3 Applications of PNIPAAm based thermally responsive copolymers............ 9

1.3.1 Reversibly switchable wettability of surface........................................ 9

1.3.2 Micro/nano-fluidic controlled transportation....................................... 10

1.3.3 Controlled drug delivery....................................................................... 10

1.3.4 Bioengineering...................................................................................... 11

1.3.5 Sensors and actuators............................................................................ 12

1.3.6 Other applications................................................................................. 12

1.4 Thesis overview............................................................................................. 14

1.5 References..................................................................................................... 16

CHAPTER 2: SYNTHESIS OF THE POLY(N-ISOPROPYLACRYL-

AMIDE-CO-HEXAFLUOROISOPROPYL ACRYLATE) COPOLYMER

VIA SURFACE-INITIATED ATOM TRANSFER RADICAL

POLYMERIZATION .......................................................................................... 25

2.1 Introduction................................................................................................... 25

2.2 Experimental.................................................................................................. 28

v
 2.2.1 Materials and chemicals....................................................................... 28

2.2.2 Synthesis procedure.............................................................................. 28

2.2.3 Characterization.................................................................................... 30

2.2.4 Water contact angle measurement........................................................ 30

2.3 Results and discussion................................................................................... 32

2.3.1 Synthesis of the poly(NIPAAm-co-HFIPA)-grafted surfaces............... 32

2.3.2 Chemical characterization of poly(NIPAAm-co-HFIPA) .................... 36

2.3.3 LCST of poly(NIPAAm-co-HFIPA) investigation............................... 37

2.4 Conclusions................................................................................................... 38

2.5 References..................................................................................................... 39

CHAPTER 3: A CHEMO-MECHANICAL SWITCH FOR

CONTROLLABLE WATER TRANSPORTATION BASED ON POLY(N-

ISOPROPYLACRYLAMIDE-CO-HEXAFLUOROISOPROPYL

ACRYLATE) COPOLYMER.............................................................................. 41

3.1 Introduction................................................................................................... 41

3.2 Experimental.................................................................................................. 43

3.2.1 Materials and chemicals....................................................................... 43

3.2.2 Synthesis procedure............................................................................. 43

3.2.3 Morphology and chemical composition characterization..................... 44

3.2.4 Contact angle measurement.................................................................. 44

3.3 Results and discussion .................................................................................. 45

3.3.1 Mechanism of chemo-mechanical switch............................................. 45

3.3.2 Effect of copolymer sequences on switching performance.................. 50

3.3.3 Reversibility and stability of switch.................................................... 52

3.3.4 Wenzel and Cassie–Baxter equation..................................................... 55

3.4 Conclusions................................................................................................... 57

3.5 References..................................................................................................... 58

vi
CHAPTER 4: SYNTHESIS OF THE POLY(N-ISOPROPYLACRYL-

AMIDE-CO-METHACRYLIC ACID) COPOLYMER VIA FREE

RADICAL- INITIATED PRECIPITATION POLYMERIZATION............... 61

4.1 Introduction................................................................................................... 61

4.2 Experimental.................................................................................................. 64

4.2.1 Chemicals and materials....................................................................... 64

4.2.2 Synthesis of poly(NIPAAm-co-MAA) copolymer............................... 65

4.2.3 Characterization.................................................................................... 65

4.3 Results and discussion................................................................................. 66

4.3.1 Synthesis of cross-linked poly(NIPAAm-co-MAA) copolymer........... 66

4.3.2 LCST of poly(NIPAAm-co-MAA) investigation................................ 69

4.4 Conclusions................................................................................................... 72

4.5 References..................................................................................................... 73

CHAPTER 5: GENERATION OF CONTROLLED MONODISPERSE

POROUS POLYMER PARTICLES BY DIPPED INKJET INJECTION...... 77

5.1 Introduction................................................................................................... 77
5.2 Experimental.................................................................................................. 80

5.2.1 Reagents and solutions......................................................................... 80

5.2.2 Equipment and procedure..................................................................... 80

5.2.3 Droplet measurement and particle characterization............................. 84

5.3 Results and discussion................................................................................... 85

5.3.1 Generation of monodisperse PSS droplets by dipped inkjet injection. 85

5.3.2 Effect of injection condition on droplet size......................................... 89

5.3.3 Porous PSS particle formation and characterization............................ 93

5.3.4 Relationship of droplet size and particle diameter................................ 101

5.3.5 Generation of HDDA particles by photopolymerization...................... 102

vii
5.4 Conclusions.................................................................................................. 105

5.5 References..................................................................................................... 106

CHAPTER 6: A THERMO-RESPONSIVE HOLLOW

MICROSPHERES FOR CONTROLLED DRUG RELEASE BASED ON

POLY(N-ISOPROPYLACRYLAMIDE-CO-METHACRYLIC ACID)

COPOLYMER...................................................................................................... 109

6.1 Introduction................................................................................................... 109

6.2 Experimental.................................................................................................. 112

6.2.1 Reagents and materials......................................................................... 112

6.2.2 Preparation of hollow core-porous shell HDDA particles................... 112

6.2.3 Synthesis of poly(NIPAAm-co-MAA) grafted HDDA microspheres.. 112

6.2.4 Particle characterization....................................................................... 113

6.2.5 Drug loading and release...................................................................... 114

6.3 Results and discussion................................................................................... 115

6.3.1 Preparation of hollow core-porous shell HDDA particles................... 115

6.3.2 Synthesis of poly(NIPAAm-co-MAA)-HDDA microspheres.............. 119

6.3.3 Drug loading and release investigation................................................ 125

6.4 Conclusions................................................................................................... 129

6.5 References..................................................................................................... 130

CHAPTER 7: GENERAL CONCLUSIONS................................................ 133

PUBLICATION LIST.......................................................................................... 137

ACKNOWLEDGMENTS.................................................................................... 139

viii
 LIST OF ABBREVIATIONS

AAO Anodic aluminum oxide

APTMS 3-Aminopropyltrimethoxysilane
ATRP Atom transfer radical polymerization
BIBB 2-bromoisobutyryl bromide
CA Contact angle
CP Cloud point
CV Coefficient of variation
DMF N,N-dimethylformamide
DOD Drop-on-demand
EDX Energy-dispersive X-ray spectroscopy
FE-SEM Field-emission scanning electron microscope
FT-IR Fourier transform infrared
HDDA 1,6-Hexanediol diacrylate
HFIPA Hexafluoroisopropyl acrylate
HPLC High-performance liquid chromatography
KPS Potassium persulfate
LCST Lower critical solution temperature
MAA Methacrylic acid
MBA Methylenebisacrylamide
NaPSS Sodium poly(styrenesulfonate)
P(AA-AM) Poly(acrylic acid-co-acrylamide )
P(DL)-HMPMA Poly(N-(DL)-(1-hydroxymethyl) propylmethacrylamide)
Poly(NIPAAm-co-HFIPA) Poly(N-isopropylacrylamide-co-hexafluoroisopropyl
acrylate)

ix
Poly(NIPAAm-co-MAA) Poly(N-isopropylacrylamide-co-methacrylic acid)
PAA Poly(acrylic acid)
PBS Phosphate buffer solution
PDEAAm Poly(N,N'-diethacrylamide)
PDMS Poly(dimethyl siloxane)
PEG Polyethylene glycol
PEtOx Poly(N-ethyl oxazoline)
Photocure-1173 2-Hydroxy-2-methylpropiophenone
PMDETA N,N,N',N",N"-pentamethyl diethylenetriamine
PMVE Poly(methylvinylether)
PNIPAAm Poly(N-isopropylacrylamide)
PVCL Poly(N-vinylcaprolactam)
QCM Quartz crystal microbalance
RAFT Reversible addition fragmentation chain transfer
SD Standard deviation
SDS Sodium dodecyl sulfate
SEM Scanning electron microscope
SWCNTs Single-wall carbon nanotubes
TCPS Tissue culture polystyrene
UCST Upper critical solution temperature
XPS X-ray photoelectron spectroscopy

x
CHAPTER 1

INTRODUCTION

1.1 Stimuli-responsive polymers

Stimuli-responsive polymers or called smart polymers are polymers mimic biological

systems in a crude way where their property change in response to external stimulus.[1]

The changed property including solubility, hydrophilic/hydrophobic balance, conformation,

degradation, surface energy, charge state, et al.[2] External stimulus can be classified into

physical, chemical, biological, and dual stimuli, as illustrated in Figure 1-1.[3-5]

Physical stimuli mainly include temperature,[6,7] light irradiation,[8,9] electric

field,[10,11] ultrasound, magnetic fields and mechanical deformation. Among them, the

temperature, light irradiation, and electric field were usually used as the stimulus for

affecting the properties of polymer structures via changing polymer/solvent system at the

energy level. Chemically dependent stimuli comprise pH,[12,13] ionic strength,[14] redox

(or electrochemical), [15,16] and solvent. They can modulate molecular interactions

between polymer and solvent molecules, or between polymer chains.[17] Biological

stimuli typically involve the actual functioning of molecules which come from the

organism, such as glucose, glutathione, enzymes, receptors, and metabolites in

inflammation.[18,19] In addition, there are dual stimuli-responsive polymers that

simultaneously respond to a combination of two or more stimulus.[20-22]

1
Figure 1-1. Classification of the stimulus of stimuli-responsive polymers. (Ref. [3]).

2
1.2 Thermally responsive poly(N-isopropylacrylamide)

1.2.1 Thermally responsive polymers

Thermally responsive polymers have attracted great attention in stimuli-responsive

polymers because of their wide application in the fields of surface modification,[23-25]

drug controlled delivery,[1,26] sensors and actuators,[27,28] biomedical materials,[29]

microfluidic devices,[30,31] chromatography,[32-35] and so on. Normally, these thermally

responsive polymers are exhibit a volume phase transition at a certain temperature, which

causes an abruptly change in the solvation state. The volume phase transition of polymers

is caused by Van-der-Waals interaction, hydrophobic interaction, hydrogen bonding with

the change in ionic interaction, and attractive ionic interaction. Critical solution

temperature is used for characterization the volume phase transition of polymers. In which,

the polymer become insoluble and a phase separation appears above a specific temperature

upon heating, possess a lower critical solution temperature (LCST). Alternatively, polymer

solutions that appear as monophasic phase above a specific temperature and biphasic

below it have the upper critical solution temperature (UCST).

Figure 1-2 shows chemical structures of typical thermally responsive polymers. The

poly(N-isopropylacrylamide) (PNIPAAm),[36,37] poly(N, N'-diethacrylamide)

(PDEAAm),[38] poly(N-(DL)-(1-hydroxymethyl) propylmethacrylamide) (P(DL)-

HMPMA),[39-41] poly(N-vinylcaprolactam) (PVCL),[27,42-44] polyethylene glycol

(PEG),[45] poly(methylvinylether) (PMVE),[46-48] and poly(N-ethyl oxazoline)

(PEtOx),[49,50] which possess the value of LCST at 32 oC, 33 oC, 37 oC, 32 oC, ＞90
o
C,

37 oC, and 62 oC, respectively. While the poly(acrylic acid-co-acrylamide) (P(AA-AM))

have an UCST of 25 oC.[51,52] Besides, several novel thermally responsive polymers have

developed over the past years.[53-55]

3
Figure 1-2. Examples of chemical structures of thermally responsive polymers.

4
1.2.2 Poly(N-isopropylacrylamide)

Poly(N-isopropylacrylamide) (PNIPAAm) is one of the most extensively studied

thermal responsive polymers which has an LCST of 32 oC in aqueous solution.[56-61] It

conformation shows a reversible transition between collapsed and swelling as the

temperature around the LCST changes. The volume phase transition of PNIPAAm caused

by alters the distribution of hydrophilic (hydrogen bonding) and hydrophobic interaction.

As indicated in the Figure 1-3, the competing hydrogen bonding between intra- and

intermolecular around LCST result in the chemical structure and conformation of

PNIPAAm changing.

At the temperature below LCST, the polymer showed highly hydrophilic

characteristics because of the dominant intermolecular hydrogen bonding between the

PNIPAAm chains and water molecules. As a result, the polymer is solubilisation in

aqueous solution. In contrast, at temperatures above the LCST, the polymer became

hydrophobic because the intramolecular hydrogen bonding between the C=O and N–H

groups of the PNIPAAm chains results in a collapsed conformation, which makes it

difficult for the hydrophilic C=O and N–H groups to interact with water molecules.

Therefore, biphasic appear in the aqueous polymer solution.

The LCST of PNIPAAm is independent of the molecular weight and the

concentration of monomer.[62] However, the LCST shifting can be achieved by

copolymerization with a hydrophilic/hydrophobic monomers, because of the LCST of

PNIPAAm is determined by the balance between hydrophilicity and hydrophobicity.

Incorporation or modification with hydrophobic comonomers can decrease the LCST,

whereas modification of the hydrophilic comonomers has the opposite effect.[63,64] In

addition, the shift effect can be influenced by the concentration of comonomers. Thus,

desired LCST or improved properties of polymers can be obtained by copolymerization

with selected and appropriate proportions of comonomers into the NIPAAm.[65-68]

5
Figure 1-3. Chemical structure and conformation of PNIPAAm changing with LCST

in aqueous solution.

6
1.2.3 Synthesis of poly(N-isopropylacrylamide)

Living free-radical polymerization (also known as reversible-deactivation radical

polymerization) approach, such as, atom transfer radical polymerization (ATRP) and

reversible addition fragmentation chain transfer (RAFT), are usually employed to

synthesise the NIPAAm or NIPAAm based copolymers.[69-73] ATRP is one of the most

useful methods for organic polymerization, which initially reported by Wang and

Matyjaszewski in 1995.[74,75] The name comes from it could employ atom transfer from

an organic halide to a transition-metal complex to generate the activity radicals, then the

transition metal transfer back to a product radical to form the final product.[76] In ATRP,

the end groups of the polymers are determined by the used initiator. Therefore, the

multifunctional polymers of different compositions and shapes, such as block copolymers

and hyperbranched polymers, can be synthesized by the ATRP method. Various NIPAAm

based copolymers were synthesized by the ATRP method for particle or membrane surface

modification.[77-81]

RAFT polymerization is the most versatile of the living free-radical polymerization

methods due to its compatibility with a wide range of functional monomers and reaction

media along with its relative ease of use.[82-84] It is providing good control over the

polymerization of vinyl esters and vinyl amides than other living free-radical

polymerization methods. Additionally, RAFT is compatible with a wide variety of reaction

media, being routinely applied in organic solution, aqueous solution and in the dispersed
phase. Therefore, an amount of NIPAAm based block copolymers were synthesized via

RAFT polymerization.[71-73,85-87]

In addition, free radical-initiated precipitation polymerization, precipitation

polymerizations [88] combined with living free-radical polymerization have been explored

for the fabrication of NIPAAm based polymer nano/microspheres.[89-91] The free

radical-initiated precipitation polymerization initiating groups on the particle nuclei would

serve as growth sites for monomer polymerization on surfaces. It takes place via radical

7
initiation of the monomers/cross-linkers in a homogeneous system followed by

propagation through a chain addition mechanism resulting in precipitation of the polymer

network. The "grafting from" mechanism, the macromolecular backbone is chemically

modified in order to introduce active sites capable of initiating functionality, in free

radical-initiated precipitation polymerization allows a precise control of polymer network

thickness. Furthermore, click chemistry [92,93] combination of other polymerization

methods, such as ATRP, RAFT, and so on, were used for NIPAAm based polymer

preparation.[94-96]

8
1.3 Applications of NIPAAm based thermally responsive copolymers

With the advantages properties of sharp range of liquid–solid phase transition,

approximately physiological temperature of LCST, hydrophilic/hydrophobic reversible

transition, adjustable LCST by copolymerization of comonomers, the PNIPAAm polymers

and it based copolymers have been used in a large variety of applications, mainly include

reversibly switchable wettability of surface, micro/nano-fluidic controlled transportation,

controlled drug delivery, bioengineering, and so on.

1.3.1 Reversibly switchable wettability of surface

Smart surfaces with reversibly switchable wettability have gained great attention

because of they are widely industrial applications, such as oil/water separation,

self-cleaning surfaces, textiles, filters.[97] Normally, the surface with reversible switching

between superhydrophobicity and superhydrophilicity can be achieved by use of

PNIPAAm grafted on the substrates with special surface morphologies.

Sun et al. reported a switchable superhydrophobic/superhydrophilic surface by

synthesis the PNIPAAm on a grooved substrate. The fabricated thermally responsive

surface exhibited a water contact angle (CA) of about 0o below 29 oC, whereas it was about

150o above 40 oC.[36] Fu and coworkers investigated the surface energy and topography of

PNIPAAm-modified anodic aluminum oxide (AAO) membranes. They found the

PNIPAAm-modified nanotextured surfaces have a great influence on macroscopic wetting

behavior, and the surface shows a similar effect as Sun's report.[37] Xia et al. developed

multi-responsive surfaces by synthesis of NIPAAm-co-PBA block polymer on silicon

substrates. The surface that can reversibly switch between superhydrophilicity and

superphobicity in response to glucose, temperature, and pH changes.[98]

9
1.3.2 Micro/nano-fluidic controlled transportation

Switchable channels that can be modulated dynamically of controlling microfluidic

even nanofluidic transportation, have attracted considerable interest recently due to their

great demand for use in microfluidic devices, molecular separation, biological system,

etc.[99-101] Grafting temperature responsive polymers to the internal surface of the

channel at the micro- or nanoscale were used to fabricate these kinds of switchable

channels.

Zhu et al. synthesized a photo-thermally sensitive PNIPAAm/graphene oxide (GO)

nanocomposite hydrogels. The combination of PNIPAAm with GO hydrogel leads to

excellent photothermal property, where the phase transition of the hydrogel can be

remotely controlled by laser exposure or non-exposure. In the study, a liquid microvalve

under the control of a near-infrared (NIR) laser was fabricated using this nanocomposite

hydrogel.[102] The similar idea was used in their latterly work, a magnetic sensitive

PNIPAAm/Fe3O4 nanocomposite hydrogel with highly effective photothermal property

were synthesized, where black Fe3O4 nanoparticles acted as a highly effective

photothermal agent.[103] A microvalve was fabricated based on the excellent photothermal

properties and it can control the fluidic flow remotely by an NIR laser. Jiang group

developed a biomimetic asymmetric responsive single nanochannel system by respectively

modifying PNIPAAM and PAA on the two side of the inner surface of the single

nanochannel. The developed system shows tunable ionic transport properties through
alternating pH and temperature control.[104]

1.3.3 Controlled drug delivery

There are many pathological areas demonstrate distinct hyperthermia. In addition,

there exist various means to heat the required area in the body so that realize the drug

release at the desired site. Thus, developing temperature responsive drug delivery systems

10
have practical clinical applications. With biocompatibility and the value of LCST at 32 °C,

PNIPAAm becomes a very interesting material for controlled release application.

Vasani et al. produced a temperature responsive inorganic-organic composite material

by graft PNIPAAm brush from porous silicon films.[105] The present composite material

has shown sustaining high drug loading and excellent control over drug release. Similar

idea have used in Szuwarzyński's work.[106] They developed a pulsatile releasing

platform that built of nanocontainers, an ordered porous AAO plate, grafted with

PNIPAAm brushes. With the temperature change around LCST of the brushes, the

opening/closing of the nanocontainers is reversible, and a pulsatile release can be easily

realized. Qian and Wu synthesis of PNIPAM semi-hollow spheres for doxorubicin

controlled release.[107] The release study and cell viability assay indicate that these

semi-hollow spheres are promising as drug release platforms with a controlled release

capability.

1.3.4 Bioengineering

Tsai et al. created a cytocompatible substrate by dip coating the PNIPAAm microgels

on polystyrene substrates, it can be used for fibroblast adhesion and temperature

responsive detachment.[108] By micropattern multifunctional PNIPAAm microgels, it can

generate complex stimuli-responsive substrates to study cell-material interactions and

allow drug delivery to cells in a spatially and temporally controlled manner. Tang et al.

developed a temperature responsive cell culture surface in a microfluidic device using

PNIPAAm grafted tissue culture polystyrene (TCPS) (PNIPAAm-TCPS).[109] The

proposed system can control cell adhesion and deadhesion by changing temperature. The

results proved that the system could be used to assess the possible interaction between cells

and PNIPAAm layer with a potential application to design a cell sheet culture surface for

tissue engineering. Li and coworkers reported a PNIPAAm-based thermo-sensitive

hydrogel containing single-wall carbon nanotubes (SWCNTs) for stem cell transplantation
in myocardial repair.[110] They found the PNIPAAm/SWCNTs hydrogel shown higher

11
bioactivities to encapsulated cells compared with PNIPAAm in vitro. And it significantly

enhanced the engraftment and survival of seeding cells in infarct myocardium and

augmented their therapeutic efficacies after myocardial infarction. Their research offered a

new perspective in development or improvement of cardiac tissue engineering scaffold.

1.3.5 Sensors and actuators

Zhang et al. fabricated a reversible, thermally- and optically responsive actuator

system utilizing composites of PNIPAAm loaded with single-walled carbon

nanotubes.[111] The present actuator is viable for many optically triggered applications.

Matsuguchi et al. created a quartz crystal microbalance (QCM) based gas sensing for HCl

dection.[112] The quartz resonator coated with PNIPAAm nanoparticles can absorb HCl

gas reversibly. The QCM-based PNIPAM nanoparticle sensor showed excellent

reproducibility and the sensitivity to 1 ppm of HCl was approximately 3.8 Hz/ppm. Richter

et al. designed two types of PNIPAAm hydrogel based actuators which can be used to

microvalves and micropumps in the microfluidic devices.[31] The first design used the

hydrogel as an actuator to close a channel by pressure generated from the swelling of the

gel, pressing on the PDMS membrane, effectively blocking the flow. The second design

used the swelling and shrinking process of hydrogel actuator to generate liquid flow.

1.3.6 Other applications

Javey group developed a thermo-responsive chemical connector based on

core/multishell hybrid nanowire fastener forests with an outer shell of PNIPAAm.[113]

The reported connector can reversibly change their wet adhesion strength around 170 times

in response to a water temperature change of less than 5 oC. He et al. synthesized a

homeostatic hydrogel material with chemo-mechano-chemical self-regulation behavior by

NIPAAm.[114] With the temperature oscillations arising from different exothermic

reactions, the material can be used for chemical reaction control. Dong and coworkers

designed an adaptive liquid microlenses by stimuli-responsive hydrogels which included

12
NIPAAm and two other pH-sensitive hydrogels.[115] The microlenses which have a focal

length ranging from -∞ to +∞ (divergent and convergent) as the human eyes work mode, is

facility to integration with existing microfluidic components or used in medical diagnostics.

Kanazawa et al. developed a new method for HPLC, name as temperature responsive

chromatography, in which uses packing materials modified with NIPAAm.[116-118] The

surface properties and functions of the stationary phases are controlled by the external

temperature, and the separation of target substances was controlled by changing the

column temperatures. It has high potential and versatility in the field of HPLC.[119,120]

13
1.4 Thesis overview

The main content of this dissertation can be divided into two parts. Part one was

focused on utilizing the hydrophilic/hydrophobic reversible transition property of

PNIPAAm to design a smart switch for water controlled transportation. It includes the

Chapter 2 and Chapter 3. While the second part emphasized the use of the switchable

conformation (swelling/collapsed) and adjustable of LCST of PNIPAAm to fabricated a

temperature responsive drug delivery for controlled drug release. It includes the Chapter 4,

Chapter 5 and Chapter 6.

In part one, regarding the individual PNIPAAm was barely hydrophobic even the

temperature above the LCST, the hydrophobic comonomer was added to obtain the real

sense of hydrophilic/hydrophobic transition polymers. As described in Chapter 2, the

poly(N-isopropylacrylamide-co-hexafluoroisopropyl acrylate) (poly(NIPAAm-co-HFIPA))

copolymer was initially synthesized on glass plate surface via surface-initiated atom

transfer radical polymerization (ATRP). The comonomer HFIPA, which was

water-resistant, was added to enhance the hydrophobicity of the polymer. The water

contact angle (CA) results were demonstrated that the copolymer

poly(NIPAAm-co-HFIPA) was shown the real sense of hydrophobicity to water at above

LCST, and shown hydrophilicity at below LCST. Based on this work, a smart and

reversible chemo-mechanical switch was developed for controllable water transportation

(Chapter 3). The smart switch was constructed by grafting poly(NIPAAm-co-HFIPA) on a

commercial multi-capillary structured plate. Water controllable transportation was realized

by switchable surface energy and the hydrophilic/hydrophobic of the copolymer brush

could be modulated with temperature, and the capillary effect was provided by the unique

architectural structure of the capillary plate. As a result, the designed chemo-mechanical

switch exhibited excellent ―ON-OFF‖ behavior for water transportation with the
temperature changing around LCST.

14
 In part two, thermo-responsive hollow microspheres for controlled drug release were

developed. Firstly, the thermo-responsive copolymer poly(N-isopropylacrylamide-co-

methacrylic acid) (poly(NIPAAm-co-MAA)) was synthesized via free radical-initiated

precipitation polymerization (Chapter 4). The MAA was added to raise the LCST of

copolymer match to the human physiological temperature. After the optimization of the

ratio of MAA in the copolymer, the results were shown the copolymer with the NIPAAm :

MAA molar ratio of 100:10 have a satisfactory LCST of 37.5 °C. Then, the drug carrier

candidate, monodisperse hollow polymer microspheres, was proved can be prepared by

inkjet technology in a continuous process (Chapter 5). The chemical and physical

properties stability of 1,6-hexanediol diacrylate (HDDA) particles were produced by the

combination of ink-jetting and UV polymerization approach. Finally, the

thermo-responsive drug delivery system was fabricated by grafting the copolymer

poly(NIPAAm-co-MAA) on the surface of HDDA particle (Chapter 6). The drug

molecules can be enclosed when the temperature is below 37 °C, when the pore channel

was closed by the swelling copolymer. While as the temperature increasing, the

copolymers become collapsed, and the drug could release from the opening pore channel.

And the drug controlled release behavior has been demonstrated.

15
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23
24
CHAPTER 2

SYNTHESIS OF THE POLY(N-ISOPROPYLACRYLAMIDE-CO-

HEXAFLUOROISOPROPYL ACRYLATE) COPOLYMER VIA
SURFACE-INITIATED ATOM TRANSFER RADICAL
POLYMERIZATION

2.1 Introduction

PNIPAAm is an excellent thermal responsive polymer with a lower critical solution

temperature (LCST) in the water of 32°C. Its conformation shows a reversible transition

between collapsed and swelling as the temperature around the LCST changes. This effect

can be attributed to competition between inter- and intramolecular hydrogen bonds. When

the temperature is below the LCST, the intermolecular hydrogen bonds between the

PNIPAAm chains and the water molecules contribute to an extended brush structure

displaying hydrophilicity. When the temperature is above the LCST, intramolecular

hydrogen bonds in the PNIPAAm chains could lead to a compact and collapsed
conformation with the hydrophobic property.[1] A wide variety of approaches has been

developed based on this specific hydrophilicity/hydrophobicity property.[2-9]

However, as pointed out by Pelton, the NIPAAM is never hydrophobic.[10]

Tmperature dependences of water contact angle (CA) measurements on a flat glass plate

demonstrated that the individual PNIPAAm brush was barely hydrophobic even the

temperature was above the LCST, in which, the CA was 82.6±1.8°(see Figure 2-1). The

25
Figure 2-1. Temperature dependences of water contact angle for the PNIPAAm brush

on a flat glass plate.

26
experimental results proved that the NIPAAm cannot achieve real the sense of hydrophobic

what name as the water CA on the flat plate should be more than 90°.

Atom transfer radical polymerization (ATRP) is become one of the most useful

methods for organic polymerization, since it initially reported by Wang and Matyjaszewski

in 1995.[11,12] The method could be polymerize styrenes, (meth)acrylates and a variety of

other monomers in a controlled manner. The molecular weights of yielding polymers can

be accurately predetermined by the ratio of the concentrations of consumed monomer to

introduced initiator.[13,14] The name comes from it could employ atom transfer from an

organic halide to a transition-metal complex to generate the activity radicals, then the

transition metal transfer back to a product radical to form the final product.[15] In ATRP,

the end groups of the polymers are determined by the used initiator. Therefore, the

multifunctional polymers of different compositions and shapes, such as block copolymers

and hyperbranched polymers, can be synthesized by the ATRP method.

In this Chapter, a hydrophobic/hydrophilic switchable copolymer poly(N-

isopropylacrylamide-co-hexafluoroisopropyl acrylate) (poly(NIPAAm-co-HFIPA)) was

initially synthesized on glass plate surface via surface-initiated ATRP. Herein, the

copolymer poly(NIPAAm-co-HFIPA) was prepared using PNIPAAm, which governs the

thermal properties, and HFIPA, which was water-resistant, was added to enhance the

hydrophobicity of the polymer. From the results of water CA, it demonstrated that the

copolymer poly(NIPAAm-co-HFIPA) was shown hydrophobicity to water at above LCST,

and shown hydrophilicity at below LCST.

27
2.2 Experimental

2.2.1 Materials and chemicals

Quartz glass plates (1.0 mm thick), 18 mm × 18 mm in size were purchased from

Matsunami Glass Ind., Ltd (Tokyo, Japan). N-isopropylacrylamide (NIPAAm) was

obtained from Kohjin Co. Ltd. (Tokyo, Japan), and recrystallized before use.

Hexafluoroisopropyl acrylate (HFIPA), 3-aminopropyltrimethoxysilane (APTMS, 97%),

2-bromoisobutyryl bromide (BIBB, 97%), copper(I) bromide (CuBr),

N,N-dimethylformamide (DMF, 99.8%), sodium chloride (99.7%), ethanol (99%),

methanol (99.99%), sulfuric acid (98%), N, N, N′, N″, N″-pentamethyl diethylenetriamine

(PMDETA, 98%), toluene (super-dehydrated), dichloromethane (super dehydrated) and

pyridine (dehydrated) were all purchased from Wako Pure Chemical Co. (Tokyo, Japan).

For NIPAAM recrystallization, 2.0 g NIPAAm with 20 mL n-hexane was stirring at

150 rpm under 60 °C. After the NIPAAm dissolved, the solution filtration with membrane

and then left it store in the freezer for overnight. Finally, the n-hexane was filtered off and

the recrystallized NIPAAm was collected and dried under vacuum. Deionized water

obtained from a Direct-Q™5 Nihon Millipore ultrapure water system (Tokyo, Japan).

2.2.2 Synthesis procedure

Poly(NIPAAm-co-HFIPA) was synthesized using the process of surface-initiated

ATRP,[16-19] and the entire polymerization process was depicted in Figure 2-2. The glass

plate was successively sonicated for 5 min in acetone and deionized water, respectively.

The substrate was then sonicated with 0.1 M NaOH solution for 10 min to generate –OH

groups on the surface. Afterward, the substrate was washed with deionized water by

sonicated, and then dried under vacuum. Subsequently, the surface of the substrate was

aminated by treatment with a 10% v/v APTMS solution in anhydrous toluene. After

refluxing for 6 h at 130°C, excess APTMS was removed by washing with toluene and
dichloromethane. The substrate was then dried under a vacuum.

28
Figure 2-2. Synthesis of the poly(NIPAAm-co-HFIPA) copolymer brush by

surface-initiated ATRP on the glass plate surface.

29
 Subsequently, the substrate was immersed in anhydrous dichloromethane with

additional pyridine (2% v/v). After cooling the above solution to 0 °C for 1 h in the ice

bath, 2-bromoisobutyryl bromide, a polymerization initiator, was added to the solvent (0.5%

v/v). The ice bath was removed after 5 min and the reaction allowed warming to room

temperature, and the reaction was allowed to proceed for 12 h with stirring to generate the

initiator grafted surface. Afterward, the substrate was washed with acetone and toluene,

and then dried under a vacuum.

Regarding the process of PNIPAAm, the substrate was reacted with a degassed

NIPAAm solution (containing 1.0 g NIPAAm, 5 mL methanol, 5 mL DMF, 0.23 mL

PMDETA, and 0.032 g CuBr) at 60°C for 2 h. In the case of poly(NIPAAm-co-HFIPA), the

substrate was reacted with a degassed PNIPAAm solution at 60°C for 1 h, 0.4 mL of

HFIPA was then added to the solution and the reaction was allowed to proceed for another

1 h at 60°C. Finally, the polymer-grafted surfaces were washed 3 times each with

deionized water and acetone and dried under a vacuum.

2.2.3 Characterization

A scanning electron microscope combined with an X-ray energy dispersive

spectrometer (SEM-EDX, S-3400N, Hitachi, Japan) was used to chemical composition of

the glass plate surface before and after modification. X-ray photoelectron spectroscopy

(XPS) analyzes were performed on an ESCA-3400 spectrometer (Shimadzu Co., Kyoto,

Japan) equipped with a Mg Kα source (1253.6 eV) operating at 10 kV and 10 mA.

2.2.4 Water contact angle measurement

The water CA on the different substrates was measured by a self-assembled

measurement system under conditions of saturated humidity, as shown in Figure 2-3. A

thermoelectric cooler (OCE-F15P-D12, OHM Electric Co., Ltd., Japan) was used to

control the temperature. After the system temperature became stable, 5.0 μL of deionized
water was dropped carefully onto the surface of the substrates by micropipette. Images of

30
the water droplet were then obtained with a Dino-Lite digital microscope (AM7013MT,

AnMo Electronics Corporation, Taiwan). The water CA was measured by a half angle

formula based on the image of the water droplet. Measurements were made at several

different positions on each substrate, and the average of these values was determined.

Figure 2-3. Schematic illustration of the setup for water CA measurement.

31
2.3 Results and discussion

2.3.1 Synthesis of the poly(NIPAAm-co-HFIPA)-grafted surfaces

The copolymer poly(NIPAAm-co-HFIPA) was grafted onto the surface of the flat

glass plate by surface-initiated ATRP. To achieve the hydrophobicity when the copolymer

above LCST, an amount of HFIPA was added to ensure the water CA above 90°at high

temperature. The effect of the ratio of HFIPA affected on surface wettability was

investigated. As shown in Figure 2-4, the water CAs measured at low temperature and the

high temperature gradually increased with increasing ratio of HFIPA. Therefore, 20%

HFIPA was chosen as the optimum conditions which resulted in a CA of 92.3±1.7° at

40 °C and a CA of 74.5±1.3°at 25 °C.

Figure 2-4. Water contact angle on the flat glass substrate with different HFIPA ratios

of the polymer.

32
 Meanwhile, the density of the copolymer affected on surface wettability was also

investigated. The density of the polymer brush on the substrate was controlled by the time

used in amine functionalization. From XPS characterization results, which are summarized

in Table 2-1, it can be deduced the surface amination reached saturation after 4h. Herein,

the high density of the polymer brush is defined as the time used for amine

functionalization beyond 4 h; while the time for amine functionalization below 2 h

classified as low density. As indicated from the results shown in Figure 2-5, a high density

of copolymer brush was more suitable for modification. Regarding the capillary plate

modified with poly(NIPAAm-co-HFIPA), water CA experiments indicated that thermally

responsive wettability was greatly enhanced by the rough surface. Figure 2-6 shows

photographs of the water drop profile on a flat glass plate and the capillary plate at 20 °C

and 40 °C, respectively. The water CA is 74.8°at 20 °C, whereas, at 40 °C, the water CA

is 94.3°. It confirms the copolymer could change from hydrophobic to hydrophilic with

temperature change around LCST.

Table 2-1. XPS elemental analysis of the surface modified APTMS at different

reaction times.

Time (h) 0.5 1.0 2.0 4.0 8.0

N/Si ratio 1.33 1.43 1.94 2.21 2.22

33
Figure 2-5. Water contact angle on the flat glass substrate with different intensity of

polymer.

34
Figure 2-6. Photographs of the water drop profile on the flat glass plate at 20 °C and

40 °C, respectively.

35
2.3.2 Chemical characterization of poly(NIPAAm-co-HFIPA)

Energy-dispersive X-ray spectroscopy (EDX) was used to characterize the surface

chemical composites of the glass plate at each polymerization process. The results are

shown in Figure 2-7, which confirm that the copolymer film was successfully grafted on

the glass plate surface.

Figure 2-7. EDX characterization for each polymerization process. (a) Bare capillary

plate. (b) Amine functionalization. (c) Amidation. (d) Polymerization. The relative

contents of each element for each polymerization process are shown in the figure.

36
2.3.3 LCST of poly(NIPAAm-co-HFIPA) investigation

The temperature dependences of water CAs for poly(NIPAAm-co-HFIPA) modified flat

glass substrates was studied in detail and results is shown in Figure 2-8. The findings

indicate that the hydrophobic–hydrophilic transition temperature for flat glass substrates

was about 30 °C, slightly lower than that for an individual PNIPAAm. This result confirms

that the hydrophobic composition in the polymer was more conducive to intra-molecular

hydrogen bond formation.

Figure 2-8. Temperature dependences of water contact angle for

poly(NIPAAm-co-HFIPA) brush on a flat substrate.

37
2.4 Conclusion

In summary, a poly(NIPAAm-co-HFIPA) block copolymer which can be changed

between hydrophobic to hydrophilic was designed and synthesized. The water-resistant of

HFIPA was added to ensure the copolymer shows hydrophobicity at high temperature. The

hydrophobic/hydrophilic state of the designed copolymer was controlled by manipulating

the temperature of the system. The copolymer showed hydrophilic at temperatures below

20 °C, and hydrophobic at temperatures above 40 °C. It has great potential for use in

various important applications, ncluding water controllable transportation, water/oil

separation, and so on.

38
2.5 References

[1] R. Pelton, Adv Colloid Interface Sci 2000, 85, 1-33.

[2] T. Sun, G. Wang, L. Feng, B. Liu, Y. Ma, L. Jiang, D. Zhu, Angew Chem Int Ed Engl

2004, 43, 357-360.

[3] Q. Fu, G. V. Rama Rao, S. B. Basame, D. J. Keller, K. Artyushkova, J. E. Fulghum, G.

P. Lopez, J Am Chem Soc 2004, 126, 8904-8905.

[4] W. Song, F. Xia, Y. Bai, F. Liu, T. Sun, L. Jiang, Langmuir 2007, 23, 327-331.

[5] M. Szuwarzynski, L. Zaraska, G. D. Sulka, S. Zapotoczny, Chem Mater 2013, 25,

514-520.

[6] E. Amstad, S. H. Kim, D. A. Weitz, Angew Chem Int Ed Engl 2012, 51, 12499-12503.

[7] H. Ye, C. L. Randall, T. G. Leong, D. A. Slanac, E. K. Call, D. H. Gracias, Angew

Chem Int Ed Engl 2007, 46, 4991-4994.

[8] R. B. Vasani, S. J. McInnes, M. A. Cole, A. M. Jani, A. V. Ellis, N. H. Voelcker,

Langmuir 2011, 27, 7843-7853.

[9] X. Zhang, C. L. Pint, M. H. Lee, B. E. Schubert, A. Jamshidi, K. Takei, H. Ko, A.

Gillies, R. Bardhan, J. J. Urban, M. Wu, R. Fearing, A. Javey, Nano Lett 2011, 11,
3239-3244.

[10] R. Pelton, J Colloid Interface Sci 2010, 348, 673-674.

[11] J.-S. Wang, K. Matyjaszewski, J Am Chem Soc 1995, 117, 5614-5615.

[12] J.-S. Wang, K. Matyjaszewski, J Am Chem Soc 1995, 28, 7901-7910.

[13] K. Matyjaszewski, J. Xia, Chem Rev 2001, 101, 2921-2990.

[14] V. Coessens, T. Pintauer, K. Matyjaszewski, Prog Polym Sci 2001, 26, 337-377.

[15] T. E. Patten, K. Matyjaszewski, Adv Mater 1998, 10, 901-915.

[16] I. B. Malham, L. Bureau, Langmuir 2010, 26, 4762-4768.

[17] A. Chhabra, R. R. Kanapuram, T. J. Kim, J. Geng, A. K. da Silva, C. W. Bielawski, C.

H. Hidrovo, Langmuir 2013, 29, 8116-8124.

39
[18] P.-F. Li, R. Xie, J.-C. Jiang, T. Meng, M. Yang, X.-J. Ju, L. Yang, L.-Y. Chu, J

Membrane Sci 2009, 337, 310-317.

[19] T. Zhao, F.-Q. Nie, L. Jiang, J Mater Chem 2010, 20, 2176-2181.

40
CHAPTER 3

A CHEMO-MECHANICAL SWITCH FOR CONTROLLABLE

WATER TRANSPORTATION BASED ON POLY(N-
ISOPROPYLACRYLAMIDE-CO-HEXAFLUOROISOPROPYL
ACRYLATE) COPOLYMER

3.1 Introduction

Smart switches that can be modulated dynamically for controlling microfluidic

transportation, have attracted considerable interest recently due to their great demand for

use in microfluidic control, molecular separation, biological system, etc.[1-3]

Stimuli-responsive polymers grafted onto micro/nano structured channels have frequently

been utilized in developing such smart switches.[4-6] Stimuli-responsive polymers are a

category of polymers that undergo a change in their conformation, surface energy, or

charge state, triggered by an external stimulus.[7] The stimuli may come from thermal,[8,9]

pH,[10,11] light irradiation,[12,13] electric field,[14,15] or multiple factors [16,17].

PNIPAAm is one of excellent thermal responsive polymer which conformation shows

a reversible transition between collapsed and swelling as the temperature around the lower

critical solution temperature (LCST) changes. Based on this unique property, a wide

variety of applications driven by the swelling/collapse properties have been developed,

such as controlled release of drugs, actuators, chemo-mechanical oscillators, cell culture

and stripping, the immobilization of enzymes, and so on.[18-25] Most studies have mainly

41
focused on the use of PNIPAAm grafted on the substrates with special surface

morphologies, such as a grooved substrate,[18] anodic aluminum oxide (AAO)

membranes,[19] copper mesh film,[20] with the goal of achieving reversible switching

between superhydrophilicity and superhydrophobicity by enhancing wettability. However,

the design of a practical switching based on PNIPAAm for temperature controlling the

transportation of an aqueous solution still remains challenging.[26-28]

In this Chapter, a smart and reversible chemo-mechanical switch was developed for

controllable water transportation. The smart switch was constructed by integrating

poly(N-isopropylacrylamide-co-hexafluoroisopropyl acrylate) (poly(NIPAAm-co-HFIPA))

on a commercial multi-capillary structured plate. With the temperature changing around

LCST, the designed chemo-mechanical switch exhibited excellent ―ON-OFF‖ behavior for

water transportation. The necessity of HFIPA in the surface wettability transition has

demonstrated. Meanwhile, the sequence of copolymer affected on properties of switching

have mainly investigated.

42
3.2 Experimental

3.2.1 Materials and chemicals

Capillary plates (1.0 mm thick) with a pore size of 6.0 μm were purchased from

Hamamatsu Photonics (type J5022-09, Hamamatsu, Japan). N-isopropylacrylamide

(NIPAAm) was obtained from Kohjin Co. Ltd. (Tokyo, Japan) and recrystallized before

use. Hexafluoroisopropyl acrylate (HFIPA), 3-aminopropyltrimethoxysilane (APTMS,

97%), 2-bromoisobutyryl bromide (BIBB, 97%), copper(I) bromide (CuBr),

N,N-dimethylformamide (DMF, 99.8%), sodium chloride (99.7%), ethanol (99%),

methanol (99.99%), sulfuric acid (98%), N,N,N′,N″,N″-pentamethyl diethylenetriamine

(PMDETA, 98%), toluene (super-dehydrated), dichloromethane (super dehydrated) and

pyridine (dehydrated) were all purchased from Wako Pure Chemical Co. (Tokyo, Japan).

Deionized water obtained from a Direct-Q™5 Nihon Millipore ultrapure water system

(Tokyo, Japan).

3.2.2 Synthesis procedure

Poly(NIPAAm-co-HFIPA) was synthesized using the process of surface-initiated

atom transfer radical polymerization (ATRP), and the entire polymerization process as
described Chapter 2, and depicted in Figure 2-2. Briefly, the surface of capillary plate was

activity by 0.1 M NaOH solution to generate –OH groups. Afterward, it aminated by

treatment with a 10% v/v APTMS in anhydrous toluene. Then, 2-bromoisobutyryl bromide

was added to generate the initiator grafted surface. As for the polymerization, the substrate

was reacted with a degassed PNIPAM solution (containing 1.0 g NIPAAm, 5 mL methanol,

5 mL DMF, 0.23 mL PMDETA, and 0.032 g CuBr) at 60°C for 1 h, 0.4 mL of HFIPA was

then added to the solution and the reaction was allowed to proceed for another 1 h at 60°C.

Finally, the polymer-grafted surfaces were washed 3 times each with deionized water and

acetone and dried under vacuum.

43
3.2.3 Morphology and chemical composition characterization

A scanning electron microscope combined with an X-ray energy dispersive

spectrometer (SEM-EDX, S-3400N, Hitachi, Japan) was used to examine the morphology

and chemical composition of the surface of the capillary plate before and after

modification.

3.2.4 Water contact angle measurement

The water contact angle (CA) on the different substrates was measured by a

self-assembled measurement system under conditions of saturated humidity. The device

information and operation procedure was described in Chapter 2, and depicted in Figure

2-3. The water CA was measured by a half angle formula based on the image of the water

droplet. Measurements were made at several different positions on each substrate, and the

average of these values was determined.

44
3.3 Results and discussion

3.3.1 Mechanism of chemo-mechanical switch

The smart switch was constructed by integrating poly(NIPAAm-co-HFIPA) on a

commercial multi-capillary structured plate. Capillary plates which have regularly arranged

microchannel with the diameter range from a few microns to several hundred microns have

shown unique architecture and excellent optical properties.[29,30] The chemo-mechanical

switch was prepared as illustrated in Figure 3-1.

A capillary plate (1.0 mm thick) with a nominal pore size of 6.0 μm was used as a

model platform (Figure 3-1a). The copolymer poly(NIPAAm-co-HFIPA) (Figure 3-1b)

was grafted onto the surface of the capillary plate by surface-initiated ATRP. Water

controllable transportation was realized by switchable surface energy and the conformation

of the copolymer brush could be modulated with temperature, and the capillary effect was

provided by the unique architectural structure of capillary plate. At the temperatures below

the LCST, the surface of the capillary plate showed highly hydrophilic characteristics

because of the dominant inter-molecular hydrogen bonding between the PNIPAAm chains

and water molecules. As a result, water could easily penetrate through the capillary plate

via the strong capillary effect induced by the microstructures (Figure 3-1c). In contrast, at

temperatures above the LCST, the surface of the capillary plate became hydrophobic

because the intra-molecular hydrogen bonding between the C=O and N–H groups of the

PNIPAAm chains results in a shrunken conformation, which makes it difficult for the

hydrophilic C=O and N–H groups to interact with water molecules. Therefore, the

dehydrated state of the copolymer chains and the large negative capillary effect made the

switch impermeable to water, causing water to be retained on the surface (Figure 3-1d).

45
Figure 3-1. Schematic illustration of the (a) capillary plate, inset: SEM images of top

view (left) and cross-sectional view (right), (b) structure of block copolymer chain, (c)

switch at "ON" state, inset: conformation of copolymer chain, (d) switch at "OFF"

state, inset: conformation of copolymer chain.

46
 According to the capillary effect,[31-33] the water was easily driven into the capillary

when the CA was smaller than 90°. As depicted in Figure 3-2, for the hydrophobic state

(Figure 3-2a), a water contact angle θ > 90°. Unless external pressure is applied, the water

cannot permeate through the plate because of the static pressure ΔP > 0 (negative capillary

effect). For the hydrophilic state (Figure 3-2b), θ < 90°, the capillary plate cannot

withstand any pressure because the static pressure ΔP < 0 (capillary effect); and the water

can permeate the plate spontaneously.

The differential pressure, ΔP, across the meniscus, which can be described as the eq

1.[34-36]

△P = 2γLV/r (1)

where γLV is the surface tension of the liquid–vapor interface, r is the radius of meniscus.

Herein, r = d/2cos(180°–θ), so eq 1 can be reorganized as eq 2.

△P = –4γLV(cosθ)/d (2)

or eq 3

△P = –lγLV(cosθ)/A (3)

where d is the pore size of the capillary plate, l is the circumference of the pore, A is the

cross-sectional area of the pore, and θ is the advancing contact angle of liquid on the

surface.

47
Figure 3-2. Schematic diagram of the liquid wetting model of the capillary plate. (a)

Hydrophobic state. (b) Hydrophilic state. The position O denotes the center of the

spherical cap that describes the meniscus; r is the radius of the meniscus; d is the

pore size of the capillary plate．

48
 Thus, 20% of HFIPA, which was water-resistant, was added to enhance the

hydrophobicity of the surface with the objective of improving switch performance. Figure

3-3 shows photographs of water drop profile on a poly(NIPAAm-co-HFIPA) modified

capillary plate at 20 °C and 40 °C, respectively. The water CA experiments indicated that

thermally responsive wettability was greatly enhanced by the rough surface. Comparing

with the water CA on flat glass plate (Chapter 2, Figure 2-6), the water CA at 20 °C

decreased from 74.8° for a flat glass plate to 56.1° as evidenced by the Wenzel

equation;[37] whereas, at 40 °C, the water CA increased from 94.3°to 118.7°which can be

explained by the Cassie and Baxter equation.[38]

Figure 3-3. Photographs of the water drop profile on the capillary plate at 20 °C and

40 °C, respectively.

49
3.3.2 Effect of copolymer sequences on switching performance

To confirm the practical application of this switch for controllable water

transportation, a transportation process was investigated with a starting temperature of

40 °C, with a cooling down to 20 °C. As shown in Figure 3-4a, the capillary plate

modified with a random order poly(NIPAAm-co-HFIPA) brush fail to function as a switch

within a short time, as anticipated. The water was retained on the surface after 10 min

when the temperature was cooled to 20 °C, and the corresponding CA changed from 118.2°

to 113.3°. Further experiments indicated that the water penetrate through the capillary plate

within nearly 22 min. This phenomenon can be attributed to the added hydrophobicity

reagent HFIPA, which prolongs the time for transiting from intra-molecular hydrogen

bonding to inter-molecular hydrogen bonding.

To improve the switch properties, two new sequences of polymers were synthesized.

One was the synthesis PNIPAAm first, followed by grafting the block copolymer

poly(NIPAAm-co-HFIPA) (Figure 3-4b). The other sequence was the initially synthesized

poly(NIPAAm-co-HFIPA), which was then grafted on PNIPAAm (Figure 3-4c). From the

water transportation results, it can be seen that both sequences can easily realize a switch

function within 10 min when the temperature changes from 40 °C to 20 °C. However, the

sequence b is slightly superior to sequence c for transportation modulation, the entire

permeation transition cycle occurred within a shorter time (4.5±0.5 min). Therefore, a

capillary plate with sequences of b polymer brush can be used for controllable water
transportation with a high switch efficiency.

50
Figure 3-4. The effect of block copolymer sequences on switching properties. (a)

Random order of polymer. (b) First synthesis of PNIPAm then grafted on

poly(NIPAAm-co-HFIPA). (c) Initial synthesis of poly(NIPAAm-co-HFIPA) then

grafted on PNIPAm.

51
3.3.3 Reversibility and stability of switch

The temperature dependences of water CAs for poly(NIPAAm-co-HFIPA) modified

flat glass substrates and a capillary plate were studied in detail and results are shown in

Figure 3-5a. The findings indicate that the hydrophobic–hydrophilic transition temperature

for flat glass substrates and the capillary plate were nearly identical. The LCST of the

copolymer was about 30 °C, slightly lower than that for an individual PNIPAAm. This

result confirms that the hydrophobic composition in the polymer were more conducive to

intra-molecular hydrogen bond formation. Additionally, the thermally responsive

wettability of the poly(NIPAAm-co-HFIPA) modified capillary plate was greatly enhanced

by the artificial structure, consistent with previous reports.[18-20] Because of the thermally

responsive switching between hydrophilicity and hydrophobicity was related to the surface

chemical composition and surface roughness. The former provides the thermally

responsive chemical change of the surface between hydrophilicity and hydrophobicity, and

the latter enhances these properties.

To study the reversibility and stability of switching, variations in the water CAs on a

modified capillary plate when the temperature was repeatedly cycled from 40 °C to 20 °C

was investigated. In these experiments, a filter paper was placed underneath of capillary

plate. The water would be absorbed by the filter paper when it permeates to the bottom of

the capillary plate. Therefore, the water CA was transiting to 0°at the "ON" state. It can be

seen that the switch showed excellent responsiveness for more than 15 cycles (Figure
3-5b).

52
Figure 3-5. (a) Temperature dependences of water CAs for poly(NIPAAm-co-HFIPA)

brush on a capillary plate and on a flat substrate. (b) Water CA measurements of the

capillary plate modified with poly(NIPAAm-co-HFIPA) were carried out alternately

at 40 °C and 20 °C.

53
 Meanwhile, a rapid transformation between "OFF" and "ON" occurred, as a single

cycle lasts only a few minutes. The transportation process for a single cycle is shown in

Figure 3-6. Additionally, such a response persisted, even after the samples had been set

aside for at least two months, without any special protection, indicating that the switch was

mechanically and chemically stable.

Figure 3-6. Water droplet transportation process for a single cycle from 40 °C to

20 °C. (a) Images of the water droplet on the surface of capillary plate were obtained

when the system temperature was set as 40 °C. Cooling the system temperature to

20 °C and the cooling procedure required nearly 120 sec (b). (c-f) Images of the

droplet were recorded at 30 sec intervals with the temperature maintained at 20 °C.

The whole process for a single cycle was complete within 4.5 min.

54
3.3.4 Wenzel and Cassie–Baxter equation

On a smooth and chemically homogeneous surface, as illustrated in Figure 3-7 (left),

the CA, θS, is given by Young‘s equation, eq 4

cosθS = (γsv – γsl)/γlv (4)

where the γlv, γsl, and γsv are the liquid–vapor, solid–liquid and solid–vapor interfacial

tensions, respectively. When the surface is roughness, the theories that are commonly used

to correlate the surface roughness with the apparent CA of a liquid droplet on a solid

substrate are the Wenzel and Cassie–Baxter equations.

In the Wenzel case, as shown in Figure 3-7 (middle), the liquid completely fills the

grooves of the rough surface where they contact, described by eq 5

cosθW = r cosθS (5)

where θW is the apparent CA in the Wenzel mode and r is the surface roughness factor.

In the Cassie–Baxter theory, as illustrated in Figure 3-7 (right), vapor pockets are

assumed to be trapped underneath the liquid. Defining the apparent CA in the Cassie mode

as θCB, θCB can be described by eq 6

cosθCB =φs(cosθS +1) –1 (6)

where φs is defined as the solid fraction upon which the drop rests.

In this work, the surface of polymer coating capillary plate was hydrophilic when the

temperature set at 20 °C. The Wenzel mode contact was formed after the water dripped on

the surface. From eq 5, it can be found that if the CA of a liquid on a smooth surface is less

than 90°, the apparent angle on a rough surface will be smaller. Thus, the CA decreased

from 74.8°on a flat glass plate to 56.1°(capillary plate). Whereas, at 40 °C, the surface of

polymer coating capillary plate was hydrophobic that prevented the surface wetting by any

aqueous solution. Therefore, the Cassie–Baxter‘s contact mode was easily generated at the

55
initial moment when the rough and hydrophobic surface encounter water droplet. From eq

6 it can be found that the surface roughness will increase the apparent CA. So, the water

CA increased from 94.3°on a flat glass plate to 118.7°(capillary plate).

Figure 3-7. Effect of surface structure on the wetting behavior of solid substrates.

Young’s mode: a liquid drop on a flat substrate. Wenzel’s mode: wetted contact

between the liquid and the rough substrate. Cassie–Baxter’s mode: non-wetted

contact between the liquid and the rough substrate. Reference from Jiang et al.[39]

and Guo et al.[40] work.

56
3.4 Conclusion

In summary, a poly(NIPAAm-co-HFIPA) block copolymer functionalized

temperature-responsive switch for controllable water transportation was developed. The

ON/OFF state of the designed switching can be controlled by manipulating the temperature

of the system. The system showed good water permeability at temperatures below 20 °C,

and was water repellent at temperatures above 40 °C. The excellent controllability over

aqueous solution transportation indicates that it has great potential for use in various

important applications including intelligent microfluidic switching, water/oil separation,

controllable drug release, and so on.

57
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[29] N. Dudovich, D. Oron, Y. Silberberg, Nature 2002, 418, 512-514.

[30] F. Chen, S. Mao, H. Zeng, S. Xue, J. Yang, H. Nakajima, J. M. Lin, K. Uchiyama,

Anal Chem 2013, 85, 7413-7418.

[31] J. P. Youngblood, T. J. McCarthy, Macromolecules 1999, 32, 6800-6806.

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14606-14610.

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[38] A. B. D. Cassie, S. Baxter, Trans Faraday Soc 1944, 40, 546-551.

[39] X. Feng, L. Jiang, Adv Mater 2006, 18, 3063-3078.

[40] B. Wang, Y. Zhang, L. Shi, J. Li, Z. Guo, J Mater Chem 2012, 22, 20112-20.

60
CHAPTER 4

SYNTHESIS OF THE POLY(N-ISOPROPYLACRYLAMIDE-

CO-METHACRYLIC ACID) COPOLYMER VIA FREE
RADICAL-INITIATED PRECIPITATION
POLYMERIZATION

4.1 Introduction

Stimuli-responsive polymers or smart polymers can dramatically change their

properties, such as, solubility, viscoelasticity, conformation, in response to external stimuli.

They have drawn considerable interest from researchers in the field of materials science, it

is playing a vital role in many applications. Stimuli for stimuli-responsive polymers

include temperature, pH, light, and chemicals.[1-6] Temperature is the most common

stimulus for stimuli-responsive polymers,[7-12] because all physical and chemical events

refer to thermally states transfer.

PNIPAAm is the most prominent candidate of thermo-responsive polymer which has

been widely researched in the past decades. The excellent biocompatibility and the LCST

value at 32 °C makes PNIPAAm very useful for controlled drug release. The LCST of

PNIPAAm is independent of the molecular weight and the concentration of monomer.[13]

Meanwhile, the LCST of PNIPAAm is determined by the balance between hydrophilicity

and hydrophobicity. Thus, the LCST shifting can be achieved by copolymerisation with

61
hydrophilic/hydrophobic monomers. The presence of hydrophobic comonomers can

decrease the LCST, whereas hydrophilic comonomers have the opposite effect.[14,15]

Furthermore, the effect can be influenced by the concentration of comonomers.

Methacrylic acid, MAA, is a carboxylic acid which has been used extensively in the

chemical industrially. It also popularly used for the synthesis of smart polymers/copolymer

because of its hydrophilic and pH-sensitive property.[16-21] In this work, MAA was used

as a comonomer for copolymerization with NIPAAm to increase the LCST of copolymer

so that make it suitable for controlled release at the range of human physiological

temperature.

Precipitation polymerizations which was first reported by the Stöver group in

1993,[22] is a heterogeneous polymerization process in polymer science. The

polymerization initially happens as a homogeneous system in the continuous phase, where

the monomer and initiator are completely soluble. Then the insoluble polymer forms upon

initiation and thus precipitates. Afterward, the polymerization proceeds by the absorption

of monomer and initiator into the polymer particles. The process with the outstanding

advantage of the absence of any surfactant or stabilizer in the polymerization process

provides a straightforward and highly efficient protocol for the preparation of uniform and

clean polymer particles. Thus, the precipitation polymerization reaction is nearly an ideal

polymerization approach and widely used.[23-35]

Recently, "living" radical precipitation polymerization has been explored for the
fabrication of monodisperse polymer nano/microspheres. The approach is based on the

incorporation of "living" radical polymerization such as atom transfer radical

polymerization (ATRP), reversible addition fragmentation chain transfer (RAFT) or

initiator-transfer agent-terminator (iniferter) into precipitation polymerization.[36-39]

Especially, the free radical-initiated precipitation polymerization initiating groups on the

particle nuclei would serve as growth sites for monomer polymerization on the surface. It
takes place via radical initiation of the monomers/cross- linkers in a homogeneous system

62
followed by propagation through a chain addition mechanism resulting in precipitation of

the polymer network. The "grafting from" mechanism in free radical-initiated precipitation

polymerization allows a precise control of polymer network thickness.

In this Chapter, cross-linked copolymer poly(N-isopropylacrylamide-co-methacrylic

acid) (poly(NIPAAm-co-MAA)) with different comonomer composition were synthesized

via free radical-initiated precipitation polymerization. The MAA was added to adjust the

LCST of copolymer match to the human physiological temperature. After optimization of

the ratio of MAA in the copolymer, the results were shown the copolymer with the

NIPAAm : MAA molar ratio of 100:10 have a satisfactory LCST of 37.5 °C.

63
4.2 Experimental

4.2.1 Chemicals and materials

N-isopropylacrylamide (NIPAAm) was obtained from Kohjin Co. Ltd. (Tokyo, Japan)

and recrystallized from hexane before use. Methacrylic acid (MAA),

methylenebisacrylamide (MBA), and potassium persulfate (K2S2O8, KPS) were purchased

from Sigma Aldrich (St. Louis, Missouri, USA). Hexane and ethanol were obtained from

Wako Pure Chemical (Osaka, Japan). Deionized water obtained from a Direct-Q™5 Nihon

Millipore ultrapure water system (Tokyo, Japan).

Figure 4-1. Chemical structure of NIPAAm, MAA, MBA and poly(NIPAAm-co-

MAA).

64
4.2.2 Synthesis of poly(NIPAAm-co-MAA) copolymer

The poly(NIPAAm-co-MAA) copolymer was synthesized by free radical-initiated

precipitation polymerization at 70 °C using MBA as a cross-linker and KPS as an initiator.

The polymerization was carried out in a 50 mL round-bottomed three-necked flask

equipped with a reflux condenser. Different feed mole ratios of NIPAAm and MAA were

synthesized. In a typical process, 0.50 mmol of NIPAAm, 0.10 mmol of MAA and 0.05

mmol of MBA were dissolved in 25 mL boiled and deoxygenated water. Then, the mixture

was heated to 70 °C and purged with nitrogen. After the mixture stirring at 200 rpm for 30

min, 1.25 mL of 2 mg/mL KPS solution was rapidly injected. The polymerization was

carried out for 6 h with stirred at 200 rpm and kept under a nitrogen atmosphere. Finally,

the obtained copolymers were purified by repetitive cycles of centrifugation and washing

by water and ethanol.

4.2.3 Characterization

A scanning electron microscope (SEM, S-3400N, Hitachi, Japan) combined with

X-ray energy dispersive (EDX, EDAX Genesis XM4) spectrometer was used to examine

the morphology and chemical composition of copolymer. Fourier transform infrared

(FT-IR) analysis was conducted on a JASCO FT/IR-6100 spectroscopy. The cloud point

(CP) measurement (turbidimetry) was carried on by the lab fabricated devices based on an

Ocean Optics HR2000CG-UV-NIR spectrometer.

65
4.3 Results and discussion

4.3.1 Synthesis of cross-linked poly(NIPAAm-co-MAA) copolymer

Cross-linked copolymer poly(NIPAAm-co-MAA) was prepared by radical-initiated

precipitation polymerization in the presence of MBA as cross-linker and KPS as the

initiator. The mechanism of polymerization process was shown in Figure 4-2. At first, the

radical was produced from KPS, then it initiated the NIPAAm, MAA, and MBA molecules

to reaction and form poly(NIPAAm-co-MAA) copolymer. Finally, the copolymer was

precipitated from the mixture solution. In order shift the LCST of copolymer

poly(NIPAAm-co-MAA) to 37 °C, the ratio of MAA in copolymer was investigated. The

ingredients and reaction conditions for the synthesis of cross-linked

poly(NIPAAm-co-MAA) were listed on Table 4-1. The resulting copolymer were collected

by centrifugation and washing by water and ethanol. And the photograph of resulting

copolymer solution, polymer a to polymer e, at room temperature (about 25 °C) were

shown in Figure 4-3, it change from transparent to milky white with MAA concentration

increasing.

66
Figure 4-2. Polymerization mechanism of cross-linked poly(NIPAAm-co-MAA)

copolymer. (a) Radical initiators generated from KPS. (b) Polymer radicals formation
process. (c) Termination process of the free radical polymerization.

67
Table 4-1. The ingredients and reaction conditions for the synthesis of cross-linked

poly(NIPAAm-co-MAA).

Polymer NIPAAm MAA MBA KPS H2 O Temperature Time

( mmol) (mmol) (mmol) (mg/mL) (mL) (oC) (h)

a 0.5 0 0.05 0.1 25 70 6

b 0.5 0.01 0.05 0.1 25 70 6

c 0.5 0.05 0.05 0.1 25 70 6

d 0.5 0.10 0.05 0.1 25 70 6

e 0.5 0.20 0.05 0.1 25 70 6

Figure 4-3. Photograph of polymer a to polymer e at room temperature (about 25 °C).

68
4.3.2 LCST of poly(NIPAAm-co-MAA) investigation

The LCST of poly(NIPAAm-co-MAA) was influenced by various variables, such as

cross-linking agent, pH value, polymerization time, and the ratio of MAA. In this work, the

ratio of MAA was mainly studied. The CP measurement method was employed to

measurement the LCST of poly(NIPAAm-co-MAA) with varies ratio of MAA. The optical

transmittance of aqueous polymer solution at various temperatures was measured at 520

nm wavelength using UV-vis spectrometer with increasing solution temperatures. The

solution heating from 25 °C to 50 °C, and the heating rate was 1 °C per step. At each step,

the samples were stabilized for 10 min before measurements. Values for the LCST of

polymeric solutions were determined as the temperature at the inflection point in the

normalized absorbance versus temperature curve.

The results were shown in Figure 4-4, it indicated that the LCST of polymer

increased with the concentration of MAA increasing. The LCST value of sample a to e

successively were 33 °C, 35 °C, 37.5 °C, 40 °C, and 44 °C. Herein, the value of LCST for

sample a (PNIPAAm) which was corresponding to the reported value, 32-33 °C. The result

could prove the accuracy of cloud point method for LCST measurement. The increased

LCST value can be explained by the hydrophilicity of MAA molecule which enhancing the

hydrogen bonds between copolymer with water. Therefore, the MAA incorporated

copolymer need higher energy for dehydration and realize intramolecular hydrogen bonds,

so that leads to a compact and collapsed conformation with poor transmittances. However,
with the concentration of MAA increased to 0.2 mmol, in which NIPAAm was 0.5 mmol,

the minor change in transmittance was found, that means the copolymer conformation

change between swelling and collapsing become weak.

69
Figure 4-4. Typical cloud point measurement of cross-linked poly(NIPAAm-co-MAA)
in distilled water from 25 °C to 50 °C.

70
 Additionally, the real time cloud point measurement of sample c cooling from 50 °C

to 25 °C was investigated, and the result shown in Figure 4-5. In the study, 3 mL of sample

c solution at 50 °C was put into the glass cell then real time measured every 1 min until the

solution cooling down to 25 °C and transmittance value become steady. From the result, it

easily found that the whole procedure for copolymer c transition from collapsing to

swelling was about 8 min. Meanwhile, the color of copolymer solution was change from

milky white to almost transparent.

Figure 4-5. Real time cloud point measurement of cross-linked

poly(NIPAAm-co-MAA) (molar ratio is 100:10) cooling from 50 °C to 25 °C. Insert

photograph indicated the color of copolymer solution at 50 °C and 25 °C, respectively.

71
4.4 Conclusions

The cross-linked copolymer poly(NIPAAm-co-MAA) with varies ratio of MAA were

synthesized via free radical-initiated precipitation copolymerization approach. The LCST

of each copolymer was investigated. With increasing the concentration of MAA, the LCST

was increased. When the molar ratios of NIPAAm to MAA was 100: 10, an LCST of

37.5 °C was obtained, which was proximity to the human physiological temperature.

Meanwhile, the period for the copolymer conformation change was only 8 min. The

synthesized copolymer with rapid temperature responsive property has great potential for

use in human drug delivery system.

72
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276-285.

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[8] S. Park, M. Zhong, T. Lee, H. J. Paik, K. Matyjaszewski, ACS Appl Mater Interfaces

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T. Hirano, M. Tanaka, T. Niidome, Y. Katayama, T. Hirano, Y. Maeda, Langmuir 2010,

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[12] H. Kanazawa, Anal Bioanal Chem 2004, 378, 46-48.

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[19] M. G. Santonicola, G. W. de Groot, M. Memesa, A. Meszynska, G. J. Vancso,

Langmuir 2010, 26, 17513-17519.

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[25] S. Chaitidou, O. Kotrotsiou, K. Kotti, O. Kammona, M. Bukhari, C. Kiparissides,

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75
76
CHAPTER 5

GENERATION OF CONTROLLED MONODISPERSE

POROUS POLYMER PARTICLES BY DIPPED INKJET
INJECTION

5.1 Introduction

Conventional methods for polymer particle production include solid colloid

templating, suspension polymerization, spray drying, emulsion polymerization, and seeded

emulsion polymerization.[1-3] However, these techniques involve multiple steps, can be

time-consuming and a large size distribution is usually observed. All of these points narrow

the range of application for such materials. As a result, various techniques, mostly based on

the fluid dynamics, have been explored to generate uniform polymer particles in a

simplified and more straightforward manner.[4-8] Among them, microfluidic-based droplet

generation methods, in which droplets are ejected in an immiscible organic or aqueous

phase, have been proved to be the most efficient technique for producing monodisperse

polymeric microparticles.[9,10] Polymeric microparticles with controllable sizes and

shapes have been fabricated on T-junction, flow-focusing, or co-flow geometry

platforms.[11-14] Unfortunately, as clearly pointed out in some reports,[11,15,16] the

organic solvent sometimes causes poly(dimethyl siloxane) (PDMS) based microfluidic

devices to undergo swelling and contamination cannot be ignored. Thus, alternative

materials to PDMS for the microfluidic platform have been proposed to address this

77
issue.[15,17,18] However, so far, the current large scale production of monodisperse

microspheres utilizing microfluidic or other apparatus still need the accurate manipulation

technology and complicated supporting device, which should be performed by the

specialists under strictly controlled experimental conditions.

Recently, inkjet printing has attracted considerable interest in this area, because it

permits droplets to be manipulated at picoliter level for the resulting droplets to be

precisely dispersed with spatial and temporal control. Taking advantage of easily

controlling the droplet size and velocity of a fluid by changing the actuation pulse

waveform, the piezoelectric drop-on-demand (DOD) inkjet has widely used in both

industrial and scientific area settings.[19-25] In the field of particle fabrication, it initially

served as an alternative technique to spray drying technology which pioneered a new

manufacturing technique known as inkjet spray drying or jetting technology. The size and

morphology of controllable particles can be explained by a mechano-electrically governed

mechanism.[26-30] However, it should be noted that these particles were usually prepared

with the inkjet nozzle located above the solvent medium, in which the droplet is first

released into air. The particle is then formed after the droplet falls into the solvent medium.

Thus, the force of the collision between droplets and the solvent interface resulted in

particles with an irregular morphology. Additionally, the size of particles was found to be

in a narrow adjustable range which resulted from the limited injection conditions for

ink-jetting in air. To my knowledge, the generation of porous polymer particles with

well-defined sphere morphology by inkjet technology has never been reported.

In this Chapter, straightforward approaches based on dipped inkjet injection

technology for the preparation of two kinds of monodisperse porous polymer particles are

reported. Initially, a DOD inkjet injector with its head immersed in an organic continuous

phase was used to generate monodisperse sodium poly(styrenesulfonate) (NaPSS)

microparticles. By integrating the inkjet technique with ―droplet-to-particle‖ synthesis
approaches,[31-33] uniform porous PSS particles were fabricated in a one-step procedure,

78
within a few minutes. The diameters of particles could be precisely controlled by changing

the waveform exerted on the inkjet, thus producing particles with diameters in the range of

15-60 μm with a coefficient of variation (CV) in the range of 2.7-4.8%. It was also possible

to produce hollow porous polymer particles by simply decreasing the concentration of the

polymer solution. Furthermore, a chemical and physical properties stability of

1,6-hexanediol diacrylate (HDDA) particles were produced by combined inkjet injection

with UV polymerization approach. Uniform droplets of HDDA polymer solution were

ejected from an inkjet, which the nozzle was placed at the bottom position of the

continuous phase. The subsequent UV irradiation and solvent extraction resulted in the

formation of porous polymer microspheres. Particles with a narrow size distribution could

be obtained using the present system in a continuous process. The results demonstrate the

merit of the inkjet-based method for the generation of monodisperse porous polymer

particles. The method has the potential for producing functional polymer particles via the

incorporation of functional or smart materials.

79
5.2 Experimental

5.2.1 Reagents and solutions

For PSS particle formation: Sodium poly(styrenesulfonate) (NaPSS, 30 wt% in water)

with an average molecular weight of 70 kg/mol was purchased from Sigma-Aldrich (St.

Louis, Missouri, USA). Sodium dodecyl sulfate (SDS) and ethanol were purchased from

Wako Pure Chemical (Osaka, Japan). Serial dilutions of NaPSS (2.5 wt%, 5 wt%, 7.5 wt%,

10 wt% and 15 wt%) containing 10 mM SDS were used as the polymer solutions.

1-Butanol was obtained from Kanto Chemical (Tokyo, Japan) and was used as the

extraction solvent.

For HDDA particle formation: 1,6-hexanediol diacrylate (HDDA) and 2-hydroxy-2-

methylpropiophenone (Photocure-1173) were purchased from Sigma Aldrich (St. Louis,

Missouri, USA). SDS, anisole, and ethanol were obtained from Wako Pure Chemical

(Osaka, Japan). Deionized water obtained from a Direct-Q™5 Nihon Millipore ultrapure

water system (Tokyo, Japan).

5.2.2 Equipment and procedure

For PSS particle formation: The equipment used for producing porous PSS particles

formation is illustrated in Figure 5-1b. It is composed of a piezoelectric DOD inkjet

microchip (Fuji Electric, Tokyo, Japan) and an extraction solvent-filled glass cell. The

nozzle of the inkjet microchip was immersed in the extraction solvent (about 3 mm depth

from the tip). As described in our previous reports,[34,35] the inkjet microchip with a

nozzle diameter of 80 μm was controlled by a laboratory-fabricated circuit and software.

The polymer solution loaded in the inkjet microchip was pressed by the bent piezoelectric

ceramic, and the droplets were then ejected from the nozzle into the extraction solvent.

Afterward, the droplets were gradually transformed into porous particles with the water

molecules in polymer solution diffusing into the extraction phase. Finally, all particles sank
down on the bottom of the glass cell.

80
Figure 5-1. (a) Schematic illustration of the experimental set-up designed to produce

monodisperse porous polymer particles. The enlarged diagram indicates the porous
polymer particle formation mechanism in the extraction solvent. (b) The digital

photograph of the experimental set-up. The enlarged photograph is the channel

structure of inkjet microchip. (c) Typical photograph of monodisperse polymer

solution droplets jetting into the extraction solvent, droplets sizes ≈ 180 µm. The

concentration of NaPSS was 15 wt%. All procedures were performed at 25 oC.

81
 For HDDA particle formation: The equipment for HDDA particles formation was

constructed as illustrated in Figure 5-2. It composed of a piezoelectric DOD inkjet

microchip (Fuji Electric, Tokyo, Japan), a continuous phase-filled glass cell, and a UV

light source. The inkjet microchip was placed near the bottom of glass cell, and the nozzle

was immersed into the continuous phase. HDDA containing 1.0 wt% of photoinitiators

(Photocure-1173) and anisole were loaded in the inkjet microchip reservoir as the polymer

solutions for injection. The continuous phase, 10 mM of aqueous SDS solution, was

provided the places for photopolymerization and collected the resulting particles.

Herein, the density of polymer solution lighter than the continuous phase. After the

polymer solution jetting into the continuous phase-filled glass cell, it would float up and

exposes to UV irradiate. For UV-induced polymerization, a 13.6-19.2 mW at 375 nm UV

light source (NSPU510CS, Nichia Corporation, Anan, Japan) was used. Upon UV

irradiation, the particles were polymerized within a few seconds. Afterward, all particles

were deposited to the bottom of the glass cell.

All particles with continuous phase were transferred to the centrifuge tube and

repeatedly washed with water and ethanol to remove the unreacted monomer from the

sample surfaces. The resulting particles were stored in water for further characterization.

82
Figure 5-2. (a) Schematic illustration of the experimental set-up designed to produce

HDDA particles. (b) Illustration of polymer solution jetting into continuous phase

from inkjet microchip. (c) The porous polymer particle formation mechanism in the

continuous phase with UV irradiation.

83
5.2.3 Droplet measurement and particle characterization

The process of the injections was recorded on a high-speed camera (Keyence

VW-9000, Osaka, Japan). Still images were generated from the videos using a VW-9000

Motion Analyzer, and the droplet sizes in the still images were analyzed. The mean droplet

size for each injection condition was calculated from measurements of 30 droplets. An

optical microscope, Olympus system microscope BX53 equipped with a digital camera

DP73 (Olympus Co., Tokyo, Japan), was used to study the diameter of the resulting

particles. To determine the size distribution of the particles, at least 250 microspheres in

each sample were measured by the measuring software on the optical microscope. The

surface and internal morphology of particles were examined by means of a scanning

electron microscope (SEM) system (S-3400N, Hitachi Co., Tokyo, Japan). Particles were

coated with a gold layer using a sputter coater (SC-701, Sanyu Denshi Co., Tokyo, Japan)

prior to obtaining the SEM images. For the morphology of internal structures, the particles

were squeezed to split by glass plate before SEM characterization.

84
5.3 Results and discussion

5.3.1 Generation of monodisperse PSS droplets by dipped inkjet injection

The procedure employed to fabricate porous PSS microparticles combined the

generation of monodisperse droplets of polymer aqueous solution using inkjet microchip,

with subsequent mutual diffusion to form porous particles. As schematically illustrated in

Figure 5-1a, porous particles were formed by the generation of monodisperse aqueous

polymer solution droplets followed by their mutual diffusion in an organic phase. NaPSS, a

biocompatible material, has been widely used as the drug carrier in controlled-release

preparations.[36-38] Meanwhile, the high self-assembly properties of NaPSS make it

convenient for the formation of polymer particles by means of a simple process.[31]

Therefore, a NaPSS aqueous solution was selected as a model polymer solution for

producing microparticles in this work. In order to extract the water in the polymer solution

to form polymer particles, the extraction solvent should be miscible with water and

immiscible with respect to NaPSS. However, water-miscible solvents such as methanol,

ethanol and acetone, were failed to form spherical particles. The reason for this is that the

rapid solvent exchange at the interphase hindered particle formation. Consequently,

1-butanol, partially miscible in water was chosen as the extraction solvent, which permitted

robust and well-defined spherical particles to be generated.

Higher ratios of viscosity to surface tension can prevent satellite droplet production in

ink-jetting. Thus, SDS was added to the polymer solution to a final concentration of 10

mM. In this situation, the SDS not only increases fluid viscosity, but also reduces surface

tension. However, a higher viscosity would increase the difficulty of ejection and it would

be necessary to apply a larger pulse voltage to the actuator. From our experimental results,

the inkjet failed to inject when the concentration of NaPSS exceeded 15 wt%, in which the

viscosity was 6.8 mPa s. Therefore, aqueous NaPSS solutions with concentrations from 2.5

wt% to 15 wt% containing 10 mM SDS were used for droplet generation. The density,
viscosity and surface tension of each fluid is listed in Table 5-1.

85
Table 5-1. Density, viscosity and surface tension of each polymer solutions and

1-butanol.1

Solution Density (g/mL) Viscosity (mPa s) Surface tension (mN/m)

2.5 wt% NaPSS 1.009 1.690 36.77

5.0 wt% NaPSS 1.021 2.369 35.26

7.5 wt% NaPSS 1.033 3.221 34.05

10 wt% NaPSS 1.045 4.485 33.79

15 wt% NaPSS 1.071 6.822 33.59

1-Butanol 0.8098 2.581 27.97

1
All polymer solutions were containing 10 mM SDS, and the measurement temperature

control at 25 °C. The surface tension of each solution was measured using a Du Nouy

Tensiometer (Itoh Seisakusho, Ltd., Tokyo, Japan), and the viscosity was analyzed by an

Ubbelohde-type viscometer (Sibata Scientific Technology Ltd., Tokyo, Japan).

86
 Figure 5-1c shows an image of the droplet of the polymer solution ejected into

1-butanol. Monodisperse droplets were obtained one by one at a frequency of 5 droplets

per second (5 Hz). The representative single polymer droplet generation process in

1-butanol is vividly displayed in Figure 5-3 in sequential images. The polymer solution

emerged at the nozzle when the driving voltage was applied on the inkjet actuator. The

solution then moved forward to form a cylindrical pillar followed by the division of the

pillar after hundreds of microseconds. Finally, a well-defined spherical droplet was

spontaneously generated by shrinking, which induced by the surface tension. And the

droplet could remain the most stable spherical state. The entire process is complete in 2 ms

in the case of a high velocity (the initial speed was about 500 mm/s). After that, the speed

decreased, eventually reaching its sedimentation velocity. The sedimentation velocity was

detected at 2 mm/s. Since the density of the polymer solution was slightly larger than that

of 1-butanol, the droplet sank to the bottom of the glass cell by gravity.

Unlike the conventional microchip-based microfluidic approaches, a carrier phase for

droplet generation is not necessary for inkjet technology. Herein, the polymer solution is

directly injected into a continuous phase (or extraction phase) on demand, which

minimizes the risk of contamination from additional reagents. The frequency of droplet

ejection was controlled by the inkjet driver. However, due to the slow sedimentation

velocity driven by gravity, the maximum frequency of the method was 10 Hz. Moreover,

the snap-off and recoil of the polymer solution would entrainment a small amount of

extraction solvent into the nozzle of inkjet microchip. However, the entrainment effect

could be negligible in the dipped inkjet injection because the interval of each injection

(100-200 ms) was extremely short compared to the slow extraction process of solvent.

87
Figure 5-3. Photos of the aqueous polymer solution droplet generation process in

1-butanol. The injection condition set as driving voltage was 50 V with 40 μs pulse

width, droplet size ≈ 210 µm. The concentration of NaPSS was 15 wt%.

88
5.3.2 Effect of injection condition on droplet size

The sizes of droplet were mainly depending on the characteristics of the fluid, the

diameter of inkjet nozzle, and the waveform applied to the inkjet actuator. The droplet is

generated by the waveform applied to the piezo-element on an inkjet microchip. Thus,

droplet size is easily adjusted via changing the driving voltage and pulse width. Injection

conditions for monodisperse polymer droplet generation in 1-butanol were investigated

with the polymer concentration set at 15 wt%. On the other hand, the dependence of

injection condition on droplet size was fitted based on actual measurement data. As can be

seen in Figure 5-4 monodisperse droplets were generated in 1-butanol with a driving

voltage ranging from 15 V to 80 V, and with a pulse width from 15 μs to 90 μs. Herein, to

avoid damage to the inkjet microchip by a high voltage, the maximum driving voltage was

set as 80 V. As has been demonstrated in previous reports,[39] the size of the droplet

injected into air is linearly dependent on the driving voltage, but showed a more

complicated behavior with changing pulse width. A similar behavior was observed when

the droplet entered the liquid phase. Generally, the sizes of droplets increased with

increasing driving voltage and pulse width, and the size distribution ranged from 80 μm to

240 μm.

In addition, the injection conditions for polymer concentrations ranging from 2.5 wt%

to 10 wt% were also investigated (Figure 5-5). The results indicated that both injection

conditions and the adjustable range of droplet size were significantly larger than the

monodisperse droplet generated in air (Figure 5-6).

89
Figure 5-4. The scope of injection conditions for the monodisperse aqueous polymer

solution droplet generation in 1-butanol and droplet size distribution. The

concentration of NaPSS was 15 wt%. Colored regions show the possible scopes for

monodisperse droplet generation. Various colors represent the size distribution of

droplet.

90
Figure 5-5. The scope of droplet formation by inkjet injection for generating

monodisperse polymer droplets in 1-butanol. The concentration of NaPSS was (a) 2.5

wt%, (b) 5 wt%, (c) 7.5 wt%, and (d) 10 wt% (10 mM SDS). Colored regions show

the possible scopes for monodisperse droplet generation. The sizes of droplet were

ranging from 80 μm to 260 μm.

91
Figure 5-6. (a) The scope of droplet formation by inkjet injection for generating

monodisperse polymer droplets in air. Colored regions show the possible scopes for

monodisperse droplet generation. The sizes of droplet were ranging from 160 μm to

205 μm. (b) Serial photo of the aqueous polymer solution droplet ejected in air,

driving waveform; 40 V—17 μs, droplet size ≈ 175 µm. The concentration of NaPSS

15 wt% (10 mM SDS).

92
5.3.3 Porous PSS particle formation and characterization

Figure 5-7a and b shows SEM images of resulting polymer particles for a NaPSS

concentration of 15 wt%. Well-defined spherical polymer particles with a uniform size

distribution and a smooth surface were obtained. Particle formation was completed in a

single step by the ―droplet-to-particle‖ approach. The particle formation process was

triggered when the NaPSS-water droplet was injected into 1-butanol. Due to the extraction

of the water from the droplet into 1-butanol, the size of the droplet shrank and was

subsequently transformed into particles by virtue of the self-assembling property of NaPSS

molecules.

Particles with controllable diameters ranging from 20 μm to 60 μm were prepared

(Figure 5-8). The interior structures of the particles were characterized by SEM after

crushing the particle with a glass slide. Both small (20 μm) and larger diameter (30 μm)

particles showed a porous structure inside shelled surface (see Figure 5-7c and d). The

pore size is about tens to hundreds of nanometers, and the density of porosity appears to be

slightly increased toward the center of the particle.

93
Figure 5-7. SEM images of monodisperse polymer particles with the diameter of (a)

30 μm and (b) 20 μm. The interior structure of particles with diameters of (c) 30 μm

and (d) 20 μm. The concentration of NaPSS was 15 wt%. The injection condition for

(a) and (c) the driving voltage was 30 V with 30 μs pulse width, for (b) and (d) the

driving voltage was 20 V with 20 μs pulse width.

94
Figure 5-8. SEM images of polymer particles with mean diameters of (a) 26 μm, (b) 35

μm, (c) 38 μm, (d) 48 μm, and (e) 55 μm. The concentration of NaPSS was 15 wt% (10

mM SDS).

95
 The monodispersity of particles was evaluated by the mean particle size and the

diameter coefficient of variation (CV). The CV is defined by CV = (σ/µ) × 100, where σ is

the standard deviation of the diameter, and µ is the average diameter. The diameters of the

particle were measured after the resulting particle was dried overnight in a desiccator. At

least 250 particles in each sample were measured using dimensional measurement software

on the optical microscope. Figure 5-9 shows the size distribution of particles with

diameters ranging from 15 μm to 60 μm. All size categories particles have narrow size

distributions with a CV value in the range of 2.7%−4.8% (Table 5-2). The results confirm

that the monodisperse porous polymer particles were successfully prepared using the

present method. Meanwhile, from the point view of size distribution, the present method is

significantly superior to previous reports.[26,29,40]

Figure 5-9. Size distribution of the polymer particles. The concentration of NaPSS
was 15 wt%.

96
Table 5-2. Resulting particle diameters under injection setting conditions and

corresponding droplet sizes.

Injection Droplet size Particle diameter

condition Mean ± SD (μm) CV (%) Mean ± SD (μm) CV (%)
20 V—20 μs 87.4 ±4.96 5.67 21.3 ±0.59 2.78
30 V—20 μs 96.8 ±3.69 3.81 24.5 ±1.19 4.85
30 V—30 μs 123.4 ±4.03 3.27 29.2 ±1.19 4.07
40 V—30 μs 141.8 ±4.92 3.47 34.9 ±1.28 3.65
40 V—40 μs 170.8 ±3.87 2.26 41.8 ±1.42 3.39
40 V—50 μs 187.6 ±5.69 3.03 45.2 ±1.88 4.16
50 V—40 μs 206.4 ±6.24 3.02 49.8 ±2.31 4.62

97
 Hollow core−shell structures of porous particles were generated by varying the

concentration of the polymer solution. As shown in Figure 5-10, particles produced using

a concentration of NaPSS of 2.5 wt% and 5 wt% had a hollow structure. On the one hand,

the volume ratio of hollow core to shell was increased with decreasing polymer

concentration. On the other hand, the average pore size was decreased as the initial

polymer concentration was increased. Meanwhile, a dimple was found on these particles.

This is attributed to the partial collapse of the surface caused by the hollow structure. The

mechanism of for hollow core formation can be attributed to the diffusion of polymer

molecules.[41] During the generation of a polymer droplet in the extraction phase, polymer

molecules in the internal droplets diffused to the interface of the droplets and formed the

shell. As these molecules gradually being adsorbed onto the inner surface of shell, a

polymer molecule vacant area occurred in the center of droplet. And then a hollow core

structure was generated in the resulting particles.

However, the porous internal structure was generated as the concentration of the

polymer solution was increased. The reason is that the increased viscosity of the polymer

solution with a high concentration inhibited the diffusion of the polymer molecules inside

the droplet. Therefore, the internal structures became porous when the NaPSS

concentrations were higher than 7.5 wt% and 10 wt% (see Figure 5-11).

98
Figure 5-10. SEM images of surface and interior structure of particles obtain from

various polymer concentrations. The polymer concentration was (a, c) 2.5 wt% and (b,

d) 5.0 wt%. The injection condition for (a) and (c) the driving voltage was 20 V with

40 μs pulse width, for (b) and (d) the driving voltage was 20 V with 30 μs pulse width.

99
Figure 5-11. SEM images of the interior structure of a polymer particle prepared

using a concentration of NaPSS of (a) 7.5 wt% and (b) 10 wt%.

100
5.3.4 Relationship of droplet size and particle diameter

The effects of the initial droplet size and polymer concentration on the final particle

size were also investigated, and the results are shown in Figure 5-12. An approximately

linear relationship between the final particle size and the initial droplet size was observed,

and the slope of the plot varied with a change in polymer concentration. Overall, the

particle diameter was smaller than the droplet size, indicating that the volume shrinkage of

the droplets is induced by solvent diffusion. The ratios of droplet size to particle diameter

were 6.40 ± 0.14, 6.06 ± 0.17, 5.59 ± 0.08, 5.36 ± 0.13, and 4.09 ± 0.13, with the polymer

concentrations of 2.5%, 5%, 7.5%, 10%, and 15%, respectively. The results suggest that

the particle diameter can be estimated from the initial droplet size and polymer

concentration. While, the initial droplet size was depended on the waveform what applied

to the inkjet actuator. Therefore, the diameter of porous polymer particles can be easily

controlled with the approach described herein.

Figure 5-12. Effect of initial droplet size and polymer concentration on the diameter
of the final particle. The slope is indicated in the inset figure.

101
5.3.5 Generation of HDDA particles by photopolymerization

The difference of set-up for HDDA particles formation from HDDA particles

formation is the nozzle of inkjet microchip placed at the bottom of glass cell. Because the

density of HDDA is lighter than continuous phase (SDS aqueous solution), so the droplet

float up after injection. That will not be facilitating to UV polymerization. So the position

of nozzle adjusts to the bottom of glass cell. After the polymer solution jetting into the

continuous phase-filled glass cell, it would float up and exposes to UV irradiate for

photopolymerization. Figure 5-13a and b shows optical microscope and SEM images of

resulting HDDA particles at the polymer concentration of 80 wt%. The well-defined

spherical polymer particles with a smooth surface were obtained.

Figure 5-13. Optical microscope (a) and SEM images (b) of monodisperse HDDA

particles. The injection condition for (a) and (b) was the driving voltage was 40 V with
40 μs pulse width. The concentration of HDDA was 80 wt%.

102
 From the size distribution of corresponding particles result shown in Figure 5-14, it

indicated the resulting particles have a uniform size distribution with the value of CV is

2.98%. The results confirm that the monodisperse HDDA particles were successfully

prepared using the present method. Meanwhile, the hollow core−shell structures of

particles were generated by decrease the concentration of HDDA polymer solution. As

shown in Figure 5-15, the hollow core−shell structures of particles were produced using a

concentration of HDDA of 1 wt%. The interior structures of the particles were

characterized by SEM after crushing the particle with a glass slide. Herein, the frequency

of droplet formation was significantly higher than the previous PSS droplet formation, the

maximum frequency of present method was 200 Hz.

Figure 5-14. Size distribution of the HDDA particles. The data based on the Figure

4-13. Particle diameter is 56.97 ±1.70 μm, CV is 2.98%.

103
Figure 5-15. SEM images of surface (a) and interior structure (b) of HDDA particles.

The injection condition: the driving voltage was 30 V with 40 μs pulse width. The

concentration of HDDA was 1 wt%.

104
5.4 Conclusions

Two kinds of porous polymer particles with a narrow size distribution were directly

fabricated by dipped inkjet injection approach. The PSS particles with diameters in the

range of 15 μm to 60 μm could be precisely controlled by fine tuning the waveform

applied to the inkjet microchip. In addition, hollow porous PSS particles were obtained

when decreasing the concentration of polymer solution. Furthermore, the HDDA particles

were produced by combined inkjet injection with UV polymerization approach. The

cross-linked HDDA particle was stable for acid/alkali organic reagents corrosion. The

resulting particles in the micro size range represent be promising candidates for use as

functional, optical, ultrasound contrast agents, coating materials, and drug delivery system.

The smaller particle could be obtained when an inkjet with a smaller size nozzle is used.

High-throughput production could be easily achieved by fabricating a multi-nozzle inkjet

setup via adding the number of nozzles and cables. I believe that the inkjet-based approach

described here represents a potentially universal platform for the production of

monodisperse porous polymer particles, which could be carried out in the common

laboratory or industrial settings.

105
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[2] J. Tan, C. Li, J. Zhou, C. Yin, B. Zhang, J. Gu, Q. Zhang, RSC Advances 2014, 4,

13334-13339.

[3] U. Jeong, S. H. Im, P. H. C. Camargo, J. H. Kim, Y. N. Xia, Langmuir 2007, 23,

10968-10975.

[4] T. Nakashima, M. Shimizu, M. Kukizaki, Adv Drug Deliv Rev 2000, 45, 47-56.

[5] J. H. Kim, T. Y. Jeon, T. M. Choi, T. S. Shim, S. H. Kim, S. M. Yang, Langmuir 2014,

30, 1473-1488.

[6] J. T. Wang, J. Wang, J. J. Han, Small 2011, 7, 1728-1754.

[7] R. Liu, G. Ma, F. T. Meng, Z. G. Su, J Control Release 2005, 103, 31-43.

[8] X. D. He, X. W. Ge, H. R. Liu, M. Z. Wang, Z. C. Zhang, Chem Mater 2005, 17,

5891-5892.

[9] J. I. Park, A. Saffari, S. Kumar, A. Günther, E. Kumacheva, Annu Rev Mater Res 2010,

40, 415-443.

[10] D. Dendukuri, P. S. Doyle, Adv Mater 2009, 21, 4071-4086.

[11] A. S. Utada, E. Lorenceau, D. R. Link, P. D. Kaplan, H. A. Stone, D. A. Weitz, Science

2005, 308, 537-541.

[12] D. Liu, H. Zhang, B. Herranz-Blanco, E. Makila, V. P. Lehto, J. Salonen, J. Hirvonen,

H. A. Santos, Small 2014, 10, 2029-2038.

[13] T. Nisisako, T. Torii, T. Higuchi, Chem Eng J 2004, 101, 23-29.

[14] Q. Xu, M. Hashimoto, T. T. Dang, T. Hoare, D. S. Kohane, G. M. Whitesides, R.

Langer, D. G. Anderson, Small 2009, 5, 1575-1581.

[15] S. W. Choi, I. W. Cheong, J. H. Kim, Y. Xia, Small 2009, 5, 454-459.

[16] S. Takeuchi, P. Garstecki, D. B. Weibel, G. M. Whitesides, Adv Mater 2005, 17,

1067-1072.

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[17] E. Quevedo, J. Steinbacher, D. T. McQuade, J Am Chem Soc 2005, 127, 10498-10499.

[18] M. T. Gokmen, W. Van Camp, P. J. Colver, S. A. F. Bon, F. E. Du Prez,

Macromolecules 2009, 42, 9289-9294.

[19] M. Singh, H. M. Haverinen, P. Dhagat, G. E. Jabbour, Adv Mater 2010, 22, 673-685.

[20] E. B. Secor, P. L. Prabhumirashi, K. Puntambekar, M. L. Geier, M. C. Hersam, J Phys

Chem Lett 2013, 4, 1347-1351.

[21] R. D. Boehm, P. R. Miller, J. Daniels, S. Stafslien, R. J. Narayan, Mater Today 2014,

17, 247-252.

[22] F. Torrisi, T. Hasan, W. Wu, Z. Sun, A. Lombardo, T. S. Kulmala, G. W. Hsieh, S.

Jung, F. Bonaccorso, P. J. Paul, D. Chu, A. C. Ferrari, ACS Nano 2012, 6, 2992-3006.

[23] Y. Sun, X. Zhou, Y. Yu, Lab Chip 2014, 14, 3603-3610.

[24] S. Hauschild, U. Lipprandt, A. Rumplecker, U. Borchert, A. Rank, R. Schubert, S.

Forster, Small 2005, 1, 1177-1180.

[25] P. J. Smith, A. Morrin, J Mater Chem 2012, 22, 10965-10970.

[26] H. Tamura, K. Kadota, Y. Shirakawa, Y. Tozuka, A. Shimosaka, J. Hidaka, Adv

Powder Technol 2014, 25, 847-852.

[27] S. Iwanaga, N. Saito, H. Sanae, M. Nakamura, Colloid Surface B 2013, 109, 301-306.
[28] E. Tekin, P. J. Smith, U. S. Schubert, Soft Matter 2008, 4, 703-713.

[29] K. Kadota, H. Tamura, Y. Shirakawa, Y. Tozuka, A. Shimosaka, J. Hidaka, Chem Eng

Res Des 2014, 92,2461-2469.

[30] D. P. Go, D. J. E. Harvie, N. Tirtaatmadja, S. L. Gras, A. J. O'Connor, Part Part Syst

Char 2014, 31, 685-698.

[31] T. Watanabe, C. G. Lopez, J. F. Douglas, T. Ono, J. T. Cabral, Langmuir 2014, 30,

2470-2479.

[32] N. Hakimi, S. S. Tsai, C. H. Cheng, D. K. Hwang, Adv Mater 2014, 26, 1393-1398.

[33] Q. Qian, X. Huang, X. Zhang, Z. Xie, Y. Wang, Angew Chem Int Ed Engl 2013, 52,

10625-10629.

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2012, 84, 10537-10542.

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4457-4459.

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[38] J. Zhou, M. V. Pishko, J. L. Lutkenhaus, Langmuir 2014, 30, 5903-5910.

[39] N. Reis, C. Ainsley, B. Derby, J Appl Phys 2005, 97.

[40] D. P. Go, J. A. Palmer, G. M. Mitchell, S. L. Gras, A. J. O'Connor, J Biomed Mater

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108
CHAPTER 6

A THERMO-RESPONSIVE HOLLOW MICROSPHERES

FOR CONTROLLED DRUG RELEASE BASED ON POLY(N-
ISOPROPYLACRYLAMIDE-CO-METHACRYLIC ACID)
COPOLYMER

6.1 Introduction

Controlled drug delivery systems (DDS) also called smart DDS, have attracted a great

deal of attention in the field of medicine over the past few decades. Because they offer a

powerful means for delivering and releasing the therapeutics in a spatiotemporally

controlled manner, overcoming the adverse properties of conventional drug molecules, and

ultimately, improving their therapeutic efficacy and safety.[1-4] Generally, micro/nano

carriers modified with stimuli-responsive polymers were used to fabricate smart DDS. To

date, various versatile drug carriers, including liposomes, dendrimers, polymeric particles,

and inorganic particles were reported.[5-14] Among them, hollow polymeric spheres have

been adopted as a preferred candidate for drug delivery, because of the advantage of

encapsulating both of hydrophobic and hydrophilic therapeutic agents, high loading

capacity, and low production cost.[15-20] Additionally, microspheres with a uniform size

distribution have attracted great interest in drug delivery systems. Because the drug

payloads and release rate kinetics are directly dependent on both the morphology and size
of the carriers.

109
 Currently, stimulus utilized to control the behavior and properties of drug delivery

systems include pH, temperature, redox, magnetic, and ultrasonic.[21] For example,

decreased pH values (pH 6.0–7.0) usually found in pathological areas, such as most cancer

tissues, because of hypoxia and massive cell death inside the tumor. And the lower pH

value shows inside cells, especially, inside endosomes the pH value is 4.5–5.5.[22] On the

other hand, the temperature can serve as a local stimulus both within the tissue and from

the outside. There are many pathological areas demonstrate distinct hyperthermia.

Additionally, there exist various means to heat the required area in the body so that realize

the drug release at the desired site. Indeed, development of the environmentally responsive

drug carriers system by utilizing variations in pH or temperature values have been wide

studying, which have broader application.[23-27]

In this Chapter, a thermo-responsive hollow microsphere for controlled drug release

was developed based on the research achievement of Chapter 4 and Chapter 5. As shown

schematically in Figure 6-1, the monodisperse microspheres with hollow core-porous shell

structure were prepared by ink-jetting approach. Then, the thermo-responsive drug delivery

system was fabricated by grafting the surface of microspheres with copolymer

poly(N-isopropylacrylamide-co-methacrylic acid) (poly(NIPAAm-co-MAA)). Herein, the

LCST of poly(NIPAAm-co-MAA) is approximate to the human physiological temperature.

Drug loading was achieved by drug molecules gradient diffusion at the temperature above

LCST, when the pore channel opening. The drug molecules can be enclosed when the

temperature is below 37 °C, when the pore channel was closed by the swelling copolymer.

While as the temperature increasing, the copolymers become collapsed, and the drug could

release from the opening pore channel. As a proof-of-concept, the loading and release of

fluorescein in the thermo-responsive hollow microsphere were investigated. The results

demonstrated the presented microspheres have potential in the controlled drug delivery.

110
Figure 6-1. Schematic illustration of the drug transportation process by the
thermo-responsive hollow microsphere. (a) Preparation of thermo-responsive drug

delivery system, (b) Drug enclosing and releasing state with temperature changing

around LCST.

111
6.2 Experimental

6.2.1 Reagents and materials

N-isopropylacrylamide (NIPAAm) was obtained from Kohjin Co. Ltd. (Tokyo, Japan)

and recrystallized from hexane before use. Methacrylic acid (MAA), 1,6-hexanediol

diacrylate (HDDA), 2-hydroxy-2-methylpropiophenone (Photocure-1173),

methylenebisacrylamide (MBA), and potassium persulfate (K2S2O8, KPS) were purchased

from Sigma Aldrich (St. Louis, Missouri, USA). Sodium dodecyl sulfate (SDS), hexane

and ethanol were obtained from Wako Pure Chemical (Osaka, Japan). Deionized water

obtained from a Direct-Q™5 Nihon Millipore ultrapure water system (Tokyo, Japan).

6.2.2 Preparation of hollow core-porous shell HDDA particles

The equipment for HDDA particles formation was constructed as same as illustrated

in Figure 5-2, Chapter 5. The process for HDDA particle preparation was, 2.5 wt% HDDA

containing 1.0 wt% of Photocure-1173 was loaded in the inkjet microchip reservoir as the

polymer solutions for injection. The continuous phase was 10 mM of aqueous SDS

solution. After the polymer solution jetting into the continuous phase-filled glass cell, it

floating up and exposes to UV irradiate. Upon UV irradiation, the particle was polymerized.
Afterward, all particles were deposited to the bottom of the glass cell. Finally, particles

with continuous phase were transferred to the centrifuge tube and repeatedly washed with

water and ethanol to remove the unreacted monomer from the sample surfaces. The

resulting particles were stored in water for further use.

6.2.3 Synthesis of poly(NIPAAm-co-MAA) grafted HDDA microspheres

The poly(NIPAAm-co-MAA) grafted HDDA microspheres were synthesized by free

radical-initiated precipitation polymerization. The polymerization was carried out in a 50

mL round-bottomed three-necked flask equipped with a reflux condenser. Feed molar

ratios of NIPAAm and MAA at 10 to 1 was synthesized in the presence of HDDA particles.

112
Firstly, 0.50 mmol of NIPAAm, 0.05 mmol of MAA and 0.05 mmol of MBA were

dissolved in 25 mL boiled and deoxygenated water. Afterward, HDDA particles stock

solution was added. Then, the mixture was heated to 70 °C and purged with nitrogen. After

the mixture stirring at 200 rpm for 30 min, 1.25 mL of 2 mg/mL KPS solution was rapidly

injected. The polymerization was carried out for 6 h with stirred at 200 rpm and kept under

a nitrogen atmosphere. Finally, the obtained particles were purified by repetitive cycles of

centrifugation and washing with water and ethanol. The composite microspheres were

named as poly(NIPAAm-co-MAA)-HDDA.

6.2.4 Particle characterization

The processes of injections were observed through a high-speed camera (Keyence

VW-9000, Osaka, Japan). Olympus system microscope BX53 equipped with a digital

camera DP73 (Olympus Co., Tokyo, Japan), was used to investigate the size and size

distribution of particles. To determine the size distribution of the HDDA and

poly(NIPAAm-co- MAA)-HDDA particles, at least 250 microspheres in each sample were

measured by the measuring software on the optical microscope.

The surface and internal morphology of particles were examined by SEM system

(S-3400N, Hitachi Co., Tokyo, Japan). Meanwhile, the high resolution of images obtained

from a field-emission scanning electron microscope (FE-SEM, JEOL JSM-7500F).

Samples for SEM were prepared by placing a drop of washed and redispersion particle

stock solution directly onto a glass slip and drying at room temperature. All specimens for

SEM measurements were sputtered with gold layer using a sputter coater (SC-701, Sanyu

Denshi Co., Tokyo, Japan) at fixed conditions (time 150 s, current 10 mA). For the

morphology of internal structures, the particles were squeezed to split by glass plate before

SEM characterization.

Chemical compositions of the particles before and after copolymer modification were

examined by the X-ray energy dispersive spectrometer (EDAX Genesis XM4). The Raman

113
studies were carried out on a JASCO NRS-1000 Laser Raman Spectrometer, and Fourier

transform infrared spectroscopy (FT-IR) were measured by the JASCO FT/IR-6100

spectrometer.

6.2.5 Drug loading and release

The fluorescein loading is achieved by dispersion of the amount of dried

thermo-responsive hollow microspheres to the phosphate buffer solution (PBS, pH 7.4)

with 10 μg/mL of fluorescein. Then, heating the solution to 45 oC and kept 2 h, after that,

the solution was cooled to room temperature and placed 2 h. Finally, microspheres were

collected and it surface washed by water at room temperature. Fluorescein enclosed and

release studies are performed in the PBS solution with the temperature set at 37 oC and 40
o
C, respectively. The processes were recorded by the fluorescence microscopy BX53 with

an external Olympus BH2-RFL-T3 epifluorescence source.

114
6.3 Results and discussion

6.3.1 Preparation of hollow core-porous shell HDDA particles

Cross-linked HDDA particles with hollow core-porous shell were produced by the

ink-jetting approach with UV polymerization. The monodisperse polymer solution droplet

was ejected from the inkjet microchip with appropriate driving waveform. And the

polymerization occurs once the polymer droplet exposes to UV irradiate. The

polymerization process can be illustrated in Figure 6-2. At first, the radical initiators

generated from photoinitiator by irradiation of UV. Subsequently, various polymer radicals

formed through the radical initiator initiating the HDDA monomer. Finally, the

cross-linked HDDA polymer was formed by the free radical polymerization process. The

whole preparation process was completed within 1 min. Meanwhile, the resulting HDDA

particles were sunk to the bottom of the cell that easily for collection.

115
Figure 6-2. The polymerization process of HDDA. (a) Radical initiators generated

from photoinitiator Photocure-1173. (b) Polymer radicals formation process. (c)

Termination process of the free radical polymerization.

116
 As the optical micrograph shown in Figure 6-3a, the uniform microspheres were

produced by the ink-jetting approach. Herein, the condition for injection set as driving

voltage was 30 V and the pulse width was 20 μs, and the frequency was 200 Hz. The size

distribution of particles was investigated by the measuring software on the optical

microscope. Numbers of 300 particles were measured and the monodispersity of particles

was evaluated by the mean particle size and the diameter coefficient of variation (CV). The

CV is defined by CV = (σ/µ) × 100, where σ is the standard deviation of the diameter, and

µ is the average diameter. As the result indicated in Figure 6-3b, the mean diameter of

particles was 23.42 μm with the CV value of 4.56%. It's confirmed that the monodisperse

HDDA particles were successfully prepared using the present ink-jetting approach.

Meanwhile, the surface and internal morphology of particles were examined by SEM

system. A well-defined spherical particle with a hollow core structure was observed as

shown in the Figure 6-3c and d. The diameter ratio of hollow core to whole microspheres

was 80% and the corresponding volume ratio was 51%, which indicated it have the high

capacity for drug loading. The porous shell was characterized by FE-SEM and the image

was shown in Figure 6-3e and f. It can be seen, the pores with size ranging from 50 nm to

200 nm were distributed on the shell.

The mechanism of hollow core formation attributed to the diffusion of polymer

molecules, which was similar as previous hollow structures of PSS particles formation

process. Briefly, the polymer molecules in the internal droplets diffused to the interface of

the droplets after it ejected into the continuous phase. As these molecules gradually

stacking on the boundary of polymer droplet and the continuous phase, a polymer molecule

vacant area occurred in the center of droplet. And then the polymerization triggered by UV

irradiation. Finally, a cross-linked polymer shell with hollow core structure was generated

in the resulting particles. The nanoporous shell was resulting from the extraction effect and

surfactant porogenic effect. The extraction effect come from the anisole exist in the

polymer solution extracted by the water, in which the water solubility of anisole is 1.6 g/L.

117
Figure 6-3. Optical micrograph of the HDDA particles (a) and its size distribution (b).

SEM images of surface (c) and interior structure (d) of HDDA particle. Typical

FE-SEM image of the surface of HDDA particles (e, f).

118
The nanopores were generated when the anisole transported from internal droplet to the

continuous phase. On the other hand, the surfactant SDS added in the continuous phase not

only prevents droplet coalescence but also has the function of porogen reagents.[28-30]

During the hydrophobic polymer droplet formed in the continuous phase, the hydrophobic

end of SDS can insert into the polymer interface. After polymerization, the SDS molecules

with unreacted monomers were removed and the nanopores formed.

6.3.2 Synthesis of poly(NIPAAm-co-MAA)-HDDA microspheres

The thermo-responsive hollow microspheres were fabricated by grafting

P(NIPAAm-co- MAA) copolymer on the HDDA particles. The unsaturated double bonds

in the HDDA particles were crucial to formation of thermo-responsive hollow

microspheres. Herein, the unsaturated double bonds were come from the unreacted

carbon-carbon double bond at the end of HDDA molecule, see Figure 6-2c. As proved in

the previous study,[31] the concentration of unsaturated double bond in the polymer

generally depends on the polymerization time. As the FT-IR spectrum shown in the Figure

6-4a, the double peaks at 1620 cm-1 refers to unsaturated double bonds. Meanwhile, the

Raman spectrum also confirmed the unsaturated double bonds were existing on the

resulting HDDA particles, the peak at the position of 1625 cm-1 (Figure 6-4b).

Undoubtedly, as shown in the spectrum, the concentration of unsaturated double bonds was

decreased compared with the monomer. The unsaturated double bonds were mainly

attributed to the shortly UV irradiation time, within few seconds. Therefore, the copolymer
P(NIPAAm-co-MAA) could be synthesis on the surface of HDDA particles through the

unsaturated double bonds.

119
Figure 6-4. (a)FT-IR and (b) Raman spectrum of HDDA monomer and the

poly-HDDA particles after polymerization by UV irradiation. The double peak at

1620 cm-1 in FT-IR spectrum and peak at 1625 cm-1 in Raman spectrum refers to

unsaturated double bonds.

120
 Poly(NIPAAm-co-MAA)-HDDA particles were prepared by the free radical-initiated

precipitation polymerization with the monomers of NIPAAm and MAA in 10:1 molar

ratios in the presence of MBA as cross-linking agent and KPS as the initiator. Chemical

structure of the resulting poly(NIPAAm-co-MAA)-HDDA particles was confirmed by

FT-IR spectroscopy and EDX. Figure 6-5 shows the FT-IR spectrum of

poly(NIPAAm-co-MAA)-HDDA particles and HDDA particles. The main peaks are (λ

cm-1): 1725, 1630, and 1590, which represent stretch vibration of C=O, unsaturated double

bonds, and bending frequency of amide N-H, respectively. The bending frequency of

amide N-H indicated that the NIPAAm based copolymer have successfully synthesized on

the HDDA particles. Additionally, the composition of poly(NIPAAm-co-MAA) grafted

HDDA and HDDA particles were characterized by the EDX. As the result shown in Figure

6-6, the peak of nitrogen what come from NIPAAm and MBA was found in the

poly(NIPAAm-co-MAA) grafted HDDA, whereas there no nitrogen observed in the

HDDA sample.

The morphology of the poly(NIPAAm-co-MAA)-HDDA particle was examined by

FE-SEM. Figure 6-7 shows the representative FE-SEM images of the composite

microsphere. It is clearly observed that nanopores with a range of 20 nm to 100 nm still

existed on the shell of composite microsphere after copolymer grafted.

121
Figure 6-5. FT-IR spectrum of poly(HDDA) particle and poly(NIPAAm-co-MAA)

grafted HDDA particle. The peak at 1590 cm-1 refers to the bending frequency of

amide N-H.

122
Figure 6-6. EDX characterization of (a) HDDA particles and (b)

poly(NIPAAm-co-MAA) grafted HDDA particles.

123
Figure 6-7. FE-SEM images of the surface of poly(NIPAAm-co-MAA)-HDDA

particles.

124
6.3.3 Drug loading and release investigation

The capabilities of the thermo-responsive hollow microspheres for controlled drug

release were investigated by using fluorescein disodium salt. Herein, the fluorescein as

hypothetical drug molecules loaded to the microspheres by heating the mixture

fluorescein-microspheres solution 2 h under 45 oC. To simulate the human physiological

pH condition, the fluorescein was dissolved in the PBS solution with the pH value at 7.4.

The gates (pore channel) on the shell were opened by the collapsed poly(NIPAAm-

co-MAA) copolymer, and the fluorescein molecules enter to the hollow core by gradient

diffusion. Then, the fluorescein molecules are enclosed at internal of microspheres after the

temperature is below LCST (set at 37 oC), in which the gate closed by the swelling

copolymer.

The experimental setup for study of fluorescein enclosed and release was constructed

as illustrated in Figure 6-8. It composed of a fluorescence microscopy and a temperature

control system. The microspheres loading with fluorescein are dispersion in the PBS

solution at first, and then transferred to the glass tube which placed in the center of plastic

box. The plastic box was filled with water and the temperature can be maintaining or

adjusted by continuous introduction of heated water.

125
Figure 6-8. Schematic illustration of the set-up designed to investigate the loading and

release of thermo-responsive hollow microsphere.

126
 Fluorescence images of microspheres loading with fluorescein in PBS solution when

temperature keeping at 37 oC were recorded by the fluorescence microscopy. In this case,

the fluorescein molecules were enclosed at the microspheres. Compared with the naked

microspheres (without fluorescein as the blank sample) image Figure 6-9a, the green

fluorescence was obviously found in the fluorescein-microspheres as shown in the Figure

6-9b. Meanwhile, there was no significant difference fluorescence intensity were found

after the fluorescein-microspheres placed at 37 oC PBS solution for 12 h, see Figure 6-9c.

Release study carried out in PBS solution with the temperature maintained at 40 ºC and the

fluorescence images were recorded. As shown in series Figure 6-9d to f, with release time

increasing the intensity of fluorescence decreased. The results confirmed that the

microspheres could enclose and release the fluorescein with a controlled manner.

Additionally, the long time and sustained release behavior eliminated the burst release in

the conventional drug delivery system.

127
Figure 6-9. Typical fluorescence images of (a) microspheres without fluorescein,

(b)and (c) microspheres encapsulation fluorescein at the initiate and time after 12 h

under 37 oC, (d-f) fluorescein release from the microspheres under 40 oC at time of 0.5

h, 5 h, and 12 h.

128
6.4 Conclusions

In summary, thermo-responsive hollow polymeric microspheres were developed for

controlled drug release. The microspheres were fabricated by grafting the

poly(NIPAAm-co-MAA) to the cross-linked HDDA particles. The monodisperse HDDA

particles with hollow core-porous shell structure at the size of 23.42 μm were prepared by

inkjet printing combined with UV polymerization. And the LCST of the thermo-responsive

copolymer was raised to the human physiological temperature by copolymerization of

MAA. The results of fluorescein loading and release on the thermo-responsive

microspheres are confirmed it promising candidates for use in controlled drug delivery

systems.

129
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131
132
CHAPTER 7

GENERAL CONCLUSIONS

The scope of this dissertation is development of NIPAAm-based thermally responsive

block copolymers and their application for chemical switching devices. It involves four

parts research contents, polymer synthesis, smart chemo-mechanical switch fabricate,

ink-jet printing technology, and controllable drug release system.

As for the polymer synthesis, the copolymer poly(NIPAAm-co-HFIPA) was initially

synthesized on glass plate surface via surface-initiated ATRP approach. The comonomer

HFIPA was added to ensure the copolymer shows hydrophobicity to water at temperature

above LCST. As a result, the copolymer showed hydrophilic at temperatures below 20 °C,

and hydrophobic at temperatures above 40 °C. This property make it has great potential for

use in various important applications including water controllable transportation, water/oil

separation, and so on. Moreover, the other thermally responsive copolymer poly(NIPAAm-

co-MAA) with varies ratio of MAA were synthesized via free radical-initiated precipitation

copolymerization approach. The MAA was added to raise the LCST of the copolymer so

that match the human physiological temperature. With increasing the concentration of

MAA, the LCST was increased. When the molar ratios of NIPAAm to MAA was 10 : 1, an

LCST of 37.5 °C was obtained. Meanwhile, the period for the copolymer conformation

change was within 8 min. The synthesized copolymer with rapid temperature responsive
property has great potential for use in human drug delivery system.

133
 The smart chemo-mechanical switch for controllable water transportation was

constructed by integrating poly(NIPAAm-co-HFIPA) on a commercial multi-capillary

structured plate. Water controllable transportation was realized by switchable surface

energy and the hydrophilic/hydrophobic of the copolymer brush could be modulated with

temperature, and the capillary effect was provided by the unique architectural structure of

the capillary plate. The ON/OFF state of the designed switching can be controlled by

manipulating the temperature of the system. The switch showed good water permeability at

temperatures below 20 °C and was water repellent at temperatures above 40 °C. The entire

permeation transition cycle occurred within a shorter time, about 4.5 min. The excellent

controllability over aqueous solution transportation indicates that it has great potential for

use in various important applications including intelligent microfluidic switching, water/oil

separation, controllable drug release, and so on.

The ink-jet printing technology was employ to preparation of monodisperse porous

polymer particles in this dissertation. In the field of particle fabrication, ink-jet printing

initially served as an alternative technique to spray drying technology which pioneered a

new manufacturing technique known as inkjet spray drying or jetting technology. However,

generation of porous polymer particles with well-defined sphere morphology by this

technology has never been reported. In my work, two kinds of monodisperse polymer

particles were directly fabricated by dipped inkjet injection approach. The size of particles

could be precisely controlled by fine tuning the waveform applied to the inkjet microchip.

It convinced that the inkjet-based approach described here represents a potentially

universal platform for the production of monodisperse porous polymer particles, which

could be carried out in the common laboratory or industrial settings.

The controllable drug release system was fabricated by grafting the previouly

prepared poly(NIPAAm-co-MAA) onto the surface of cross-linked HDDA particle. The

monodisperse HDDA particles with hollow core-porous shell structure were prepared by
inkjet printing technology combined with UV polymerization. Drug loading was achieved

134
by drug molecules gradient diffusion at the temperature above LCST, when the pore

channel opening. The drug molecules can be enclosed when the temperature is below

37 °C, when the pore channel was closed by the swelling copolymer. While as the

temperature increasing, the copolymers become collapsed, and the drug could release from

the opening pore channel. The results of fluorescein loading and release were confirmed it

promising candidates for use in controlled drug delivery systems.

135
136
 PUBLICATION LIST

Refereed publications

*1. J Yang, D Katagiri, S Mao, H Zeng, H Nakajima, K Uchiyama, Generation of

controlled monodisperse porous polymer particles by dipped inkjet injection, RSC

Advances 2015, 5, 7297-7303.

*2. J Yang, M Hida, S Mao, H Zeng, H Nakajima, K Uchiyama, A chemo-mechanical

switch for controllable water transportation based on a thermally responsive block

copolymer, Chemical Communications 2014, 50 (71), 10265-10268.

3. J Yang, H Zeng, S Xue, F Chen, H Nakajima, K Uchiyama, Quantitative-nanoliter

immunoassay in capillary immune microreactor adopted inkjet technology, Analytical

Methods 2014, 6 (9), 2832-2836. (Inside Cover)

4. H Zeng, J Yang, D Katagiri, Y Rang, S Xue, H Nakajima, K Uchiyama, Investigation of

monodisperse droplet generation in liquids by inkjet, Sensors and Actuators B:

Chemical 2015, 220, 958-961.

5. S Xue, H Zeng, J Yang, H Nakajima, K Uchiyama, A compact immunoassay platform

based on a multicapillary glass plate, Sensors 2014, 14 (5), 9132-9144.

6. F Chen, S Mao, H Zeng, S Xue, J Yang, H Nakajima, JM Lin, K Uchiyama, Inkjet

nanoinjection for high-thoughput chemiluminescence immunoassay on multicapillary

glass plate, Analytical Chemistry 2013, 85 (15), 7413-7418.

*7. J Yang, D Katagiri, S Mao, W Zhang, H Zeng, H Nakajima, K Uchiyama, Inkjet

printing thermoresponsive core-shell polymer microcapsules for controlled drug release,

Chemistry of Materials. In preparation.

137
Academic conferences

*1. J Yang, D Katagiri, H Zeng, H Nakajima, K Uchiyama. Inkjet approach for preparation

of monodisperse porous polymer particles. Pittcon Conference & Expo 2015, Ernest N.

Morial Convention Center, New Orleans, USA, March 8-12, 2015. (Poster)

*2. J Yang, H Zeng, H Nakajima, K Uchiyama. Single-step formation of monodisperse

porous polymer microparticles with inkjet technology. RSC Tokyo International

Conference, JASIS Conference, Makuhari Messe, Japan, September 4-5, 2014. (Poster)

*3. J Yang, H Zeng, H Nakajima, K Uchiyama. Quantitatively controlled nanoliter

immunoassay utilizing inkjet technology. The 15th Beijing Conference and Exhibition

on Instrumental Analysis, Beijing, China, October 23-26, 2013. (Poster)

4. J Yang, H Zeng, H Nakajima, K Uchiyama. Nanoliter immunoassay based on capillary

immune microreactor and inkjet injection technology. RSC Tokyo International

Conference, JASIS Conference, Makuhari Messe, Japan, September 5-6, 2013. (Poster)

5. J Yang, K Uchiyama. Nanoliter immunoassay based on capillary immune microreactor

and inkjet injection technology. China-Japan-Korea Symposium on Analytical

Chemistry, Kyushu University Hospital Campus, Japan, August 22-23, 2013. (Oral
presentation)

138
 ACKNOWLEDGMENTS

First and foremost, I must express my sincere gratitude to my advisor and mentor, Dr.

Katsumi Uchiyama for providing me this precious opportunity to be part of his lab and

supporting me during the past three years. This dissertation would not have been possible

without his guidance and helpful discussions and suggestions. His patience, encouragement,

understanding, and knowledge have profoundly impacted my life as a scientist.

I would like to thank my dissertation committee members, Dr. Shoichiro Asayama, Dr.

Hizuru Nakajima, and Dr. Shungo Kato for their valuable insights and suggestions

concerning the content and presentation of my work.

I am grateful to Dr. Hulie Zeng, and Dr. Ming Yang, for their helpful suggestions and

advice in the successful completion of my research. I am sincerely appreciating for all the

advice and encouragement from Dr. Jin-Ming Lin. You make my higher education dream

come true.

I would also like to thank the staff members of Department of Applied Chemistry, Dr.

Hideki Masuda, Dr. Hirohisa Yoshida, Dr. Koichi Kajihara, and Dr. Yasuhiro Akita, for

their kind assistance in many aspects throughout my PhD program.

I am thankful to all the past and current members of Dr. Uchiyama research group for

their help, friendship and support. Especially, thanks Daisuke Katagiri, Mitsuaki Hida,

Shuhua Xue, Ying Weng, Yasuhiro Shiobara, Kazuhiro Morioka, Fengming Chen, Ying

Rang, Yuri Nakagawa, Sifeng Mao, Chiho Sato, Weifei Zhang, Hiroshi Uno, Akihito

Korenaga, Shun Kondo, Yong Zhang, Maho Asada.

Thanks to the support of Asia Human Resource Fund and Tokyo Metropolitan High

Technology Research Program.

Finally, I am thankful to all those who have contributed to the completion of my PhD

dissertation.

139