J Nanostruct 14(4): 1046-1057, Autumn 2024

RESEA RCH PAPER

Synthesis of Titanium Dioxide Nanoparticles (TiO2) and

Application for Reduction of Bacterial Growth
Mena Akram Ali *, Saba A. Mahdy, Nehia Neama Hussein

Division of Biotechnology, Department of Applied Sciences, University of Technology, Baghdad, Iraq

ARTICLE INFO ABSTRACT

The present study has been dedicated to analyzing the photocatalytic
Article History: activity and chemical stability of TiO2 nanoparticles, which in turn,
Received 02 July 2024 could be employed in environmental and medical applications, such as
Accepted 16 September 2024 antimicrobial coatings and cancer therapy. The work was focused on
Published 01 October 2024 the synthesis of titanium dioxide nanoparticles (TiO2) using titanium
tetraisopropoxide Ti[OCH(CH₃)₂]₄ dissolved in isopropyl alcohol,
Keywords: sonication, and hydrothermal method at 140°C. The nanoparticles were
Cancer afterwards cleaned, centrifuged, and dried by oven at 50°C for 3h, and
stored at refrigerator until use. In the characterization of the fabricated
Antibacterial
nanoparticles, an anatase tetragonal phase with a particle size of around 74
Antioxidant nm was established. XRD analyses (X-ray diffraction) were exploited to learn
Anti-cancer more about the nanostructure, crystallite dimensions, and crystallographic
TiO2 nanoparticles planes. Spectroscopy of TiO2 was done using UV-Vis absorbance, and the
results were in line with TiO2 photocatalytic activity which is particularly
due to its interaction with light at 385 nm. The antibacterial properties of
the nanoparticles were demonstrated in biological evaluations, and they
were found to be effective against Proteus mirabilis. This may lead to finding
out ways of using them for microbial targeting. Besides, the nanoparticles
demonstrated concentration-dependent antioxidant properties, which were
responsible for the suppression of oxidative stress. Moreover, TiO2 showed the
anti-cancer effects against Human Skin Squamous Cell Carcinoma (HSSCC)
concentrating and time-dependent cell death processes. The research,
further, studied the role of TiO2 nanoparticles in the suppression of biofilms,
especially against staphylococcal biofilms, but had a limited effect in the
species like Pseudomonas aeruginosa, suggesting that the strategies should
be specific to applications. In general, the TiO2 nanoparticles synthesized
demonstrate a wide range of applications in medicine and environment with
their antibacterial, anti-oxidant and anti-cancer properties.

How to cite this article

Ali M., Mahdy S., Hussein N. Synthesis of Titanium Dioxide Nanoparticles (TiO2) and Application for Reduction of Bacterial
Growth . J Nanostruct, 2024; 14(4):1046-1057. DOI: 10.22052/JNS.2024.04.006

INTRODUCTION
Titanium dioxide nanoparticles (TiO2 NPs) have pollutants and inactivate microbial pathogens
been acknowledged as a useful nanomaterial in when irradiated with UV light has been broadly
environmental applications due to their unique documented, which appears to be an effective
photocatalytic properties and chemical stability path to water and air purification [2]. The
[1]. The TiO2 ability to decompose organic efficiency of TiO2 nanoparticles is often an issue
* Corresponding Author Email: as.21.55@[Link] due to the recombination of the photo-generated

This work is licensed under the Creative Commons Attribution 4.0 International License.
To view a copy of this license, visit [Link]
 M. Ali et al./ Synthesis of TiO2 NPs and Application for Reduction of Bacterial Growth

electron-hole pairs and their limited absorption MATERIALS AND METHODS

in the visible light spectrum [3]. The most recent Materials
achievements in nanotechnology, including Titanium Tetra Isopropoxide (TTIP, C12H28O4Ti,
TiO2 nanoparticles, have proved to be effective 97%), Ethanol (CH3CH2OH, 96%) isopropyl
against these shortcomings. The doping of TiO2 alcohol 50 ml, and distilled water was purchased
with non-metals such as nitrogen and carbon has from Merck India. All chemicals and reagents
shown to shift the photo catalytic activity into are of analytic grade and used without further
the visible light spectrum, which increases the purification. Bacterial pathogens, such as Staph.
overall efficiency of the process [4]. Besides, the epidermidies, Staph. haemolyticus (gram-positive
synthesis of nanostructured titanium dioxide in bacteria), P. mirabilis, P. aeruginosa (gram-negative
different morphologies (such as nanotubes and bacteria) were used to study biological activities.
nanowires) has resulted in increased surface area
and improved electron transport. This has again Synthesis of TiO2 by hydrothermal method
resulted in the enhancement of photocatalytic The production of TiO2 NPs was somewhat
performance [5]. Even though these technologies altered from the previously published literature
make progress, the problems with the practical [14]. Initially, 50 milliliters of isopropyl alcohol are
use of nanostructured TiO2 are still here, and they mixed continuously for 30 minutes to dissolve 1.6
are especially about the cost-effectiveness, the milliliters of titanium tetra isopropoxide. Finally,
scalability of synthesis methods, and the long- to create the dispersion medium, add a few
term impact on the environment [6]. Facing these drops of distilled water. The product spent twenty
issues is a must for the full-scale implementation minutes in the ultrasonic bath. The solution was
of TiO2-related technologies in environmental sonicated and then placed in an autoclave set at
decontamination. 140 °C for three [Link] allowing the solution
Furthermore, TiO2 nanoparticles have a to reach room temperature, the contaminants
wide range of applications in the medical field were eliminated by centrifuging and washing
in addition to their environmental benefits. it with deionized water. Whatman No. 1 Filter
Accordingly, this type of materials inherently has paper is then used to filter it. The filtered sample
antimicrobial properties that could be employed was dried three hours at 50°C. The resulting TiO2
in the coating of medical devices and implants nanoparticles were collected and subjected to
to decrease the amount of hospital-acquired further analysis.
infections [7,8]. TiO2, the photocatalyst, is not only
able to produce self-sterilized surfaces, which are Characterization of TiO2 nanoparticles
favourable for utilization in bleached areas such The PANanalytical XPERT PRO Diffractometer
as the operating rooms, but also TiO2 is able to was used to record the titanium dioxide
produce self-sterilized surfaces [9]. nanoparticles under investigation’s X-ray diffraction
Also, TiO2 nanoparticle’s antioxidant properties pattern. Using a Perkin Elmer spectrophotometer,
have been identified, a research which led to the the FTIR spectra was acquired between 400
promotion of oxidative stress related diseases and 4000 cm−1. SEM was used to visualize the
especially cancerous diseases such as [10]. The surface morphology of TiO2 nanoparticles. The
care by the number of football players is great. homogeneity and elemental distribution of the
We have to treat the minor injuries and the chemical under investigation are ascertained
bigger injuries too [11,12]. However, despite using the EDS spectra. Quanta FEG-250 assisted
the fact that the safety and biocompatibility of in recording the SEM with the EDS spectrum. A
ns- TiO2 nanoparticles remain a challenge for Shimadzu 2700 spectrophotometer was used
nanoparticle interaction with biological systems, to record the UV-Visible Diffuse Reflectance
which may impact their toxicological effects [13]. Spectrophotometer (DRS) spectrum. The 200–
The study aims at examining the crystal structure, 800 nm range was used to record the reflectance
antibacterial, antioxidant and anticancer spectra. Using the disk diffusion technique, the
properties of chemically synthesized titanium antibacterial activity of TiO2 nanoparticles against
dioxide (TiO2) nanoparticles for possible future gram-positive and gram-negative bacteria was
medical and environmental uses. investigated.

J Nanostruct 14(4): 1057046-1057, Autumn 2024 1047

 M. Ali et al./ Synthesis of TiO2 NPs and Application for Reduction of Bacterial Growth

Antibacterial activity concentrations (200 µl per well) of the test

Mueller-Hinton agar medium was created substance. The plates were covered, incubated at
to examine the impact of titanium dioxide 37°C, and exposed for different durations (24, 48,
nanoparticles on the development of bacterial 72 hrs).
isolates. The McFarland tube was used to create
and compare the bacterial solution used in this Minimum Inhibitory Concentration
investigation. The plates containing the few Microtiter Plate Method
selected strains were cultured and left to dry at The antibacterial activity of was assessed using
the open air at room temperature. After properly minimum inhibitory concentration MIC assays
sterilizing the wells, wells were then punched against Gram-negative [Link], [Link],
into agar. For the purpose of this assay, 80μL of and [Link] [18] Stated that the MIC was
different concentrations of three type of TiO2NPs determined on a 96-well microtiter plate using
(20, 40, 60, 80, and 100)μgmL-1 for each bacterial the resazurin assisted microdilution technique
plate. The plates were incubated for twenty- in Mueller-Hinton broth (MHB) as follows: Test
four hours at 37 °C, and diameter of the cleared material, which was: Plant extract [Link] ,
zone against the microorganism was measured in TiO2 nanoparticle, Preparation of test materials
millimeters [15,16]. with the final required concentration. 100 μl of
broth medium in each well from 1 to 10 were
Anticancer Activity made.
Preparation of Cancer Cell Lines 100 μl of diluted test material was transferred
The cytotoxic effects of TiO2-NPs, The efficacies to the first well. by transferring 100 μl from the
of test sub-stances at specified concentrations first to the 10th well (the concentrations were
(Control, 1. 95, 3. 9, 7. 8, 15. 62, 31. 25, 62. 5, 100, 50, 25, 12.5, 6.25, 3.12, 1.56, 0.78, 0.39, and
125, 250, 500, 1000 µg/mL) were evaluated on 0.19 μg/ml) in addition to the controls.
a HSSCC line in passage. The cells were cultured Each well was inoculated with 100 μl of bacterial
in RPMI-1640 medium supplemented with 10% suspension equivalent to McFarland standard no
Fetal Calf Serum (FCS). The cytotoxic impact of 0. 5 (1.5 ×10 8 CFU\ml).
the test substance was examined by culturing The microtiter plate was incubated at 37 C for
cells in tissue culture plates (96-Microtiter plates) 24 h. 8- 30 μl of resazurin dye was added to each
with a flat bottom. The experiment consisted well (30 μl/well), and incubated for 2 to 4 hrs. for
of three stages: Cells Seeding: After activating the observation of color change. After completion
and proliferating cancer cell lines for 24 hr, the of the incubation, rows with no color change (blue
monolayer growth was treated with Trypsin- resazurin color remained unchanged) were scored
Versen solution. Subsequently, 25 mL of RPMI- as above the MIC whereas the last blue well in the
1640 medium, prepared with serum, was added to row recorded as MIC.
each well, adjusting the cell count to 1x104 using
a counting chamber. A volume of 100 µl of the cell Anti-biofilm
suspension was distributed into the tissue culture The microtiter plate (MTP) assay is a qualitative
wells, which were then incubated at 37 °C for 24 hr technique that uses a microplate reader to
to allow cell attachment to the glass [17]. determine an agent’s effectiveness against biofilm
formation. The minimum inhibition concentrations
Preparation of TiO2-NPs concentrations MIC obtained from the previous experiment were
Different concentrations of the test substance used to study the effect of the test materials on the
were prepared using a serum-free tissue culture formation or inhibition of biofilm of the studied
medium. These concentrations were added to [Link], [Link],and [Link]
the wells containing adherent cancer cells, and isolates that produce strong biofilm, the test
solutions were prepared just before use. Six materials were: Plant extraction ([Link] ,
replicates were used for each treatment. The TiO2NPs.
culture medium in the plates was poured out, The same previously mentioned protocol
designating column 1 as the negative control, (MTP) was used for the biofilm formation assay.
to which 200 µl of serum-free culture medium However, 100 μl of test compounds was added.
was added. Columns 2 to 12 received increasing the plate was incubated at 37 °C for 24h. After

1048 J Nanostruct 14(4): 1057046-1057, Autumn 2024

 M. Ali et al./ Synthesis of TiO2 NPs and Application for Reduction of Bacterial Growth

that, all wells were washed, stained, and read at Ascorbic acid or vitamin C at a concentration of
600 nm wavelength using a micro-plate reader. 1/1 (1 mg/100 ml of distilled water) is used as a
percent of biofilm inhibition was calculated by the positive control due to its high effectiveness as an
equation [19]: antioxidant and is considered a standard material
for comparison.
% Biofilm inhibition = [(OD Control- OD
Sample)/OD Control] ×100 Statistical Analysis
One-way analysis of variance (ANOVA) and
Antioxidant subsequent post-hoc tests were conducted using
The method mentioned in [20] was followed IBM SPSS Statistics for Windows, version 26 (SPSS
in conducting the antioxidant antioxidant Inc., Chicago, Illinois, United States). Variables
test, using the method (2, 2-diphenyl-1-picryl- were expressed as mean ± standard deviation
hydrazylhydrate DPPH), by adding 0.024 grams of (SD). The level of significance was set at 𝑝≤0.05.
DPPH to 50 milliliters of absolute ethyl alcohol. It
is dissolved well by mixing it on a magnetic stirrer RESULTS AND DISCUSSION
without heat, then the volume is supplemented X-Ray Diffraction
to 100 milliliters with absolute ethyl alcohol to Fig. 1, shown, plots intensity against 2Theta
give a final concentration equal to 0.024 mg/ml. angles, revealing crucial information about
Then, half a milliliter of serial concentrations of the crystalline structure of TiO₂. Peaks labeled
the test substance (0.0, 25, 50, 100, 150, 200) µg/ with Miller indices (such as 110, 101, and 200)
mL were taken and added to a mixture of DPPH correspond to specific crystallographic planes
(0.5.) mM and mL (3.3) of absolute ethanol. The within the material. The X-ray diffraction (XRD)
amount of color change was measured using analysis revealed the presence of nano-sized
The spectrophotometer was at a wavelength particles in the tetragonal anatase phase of TiO₂.
of 515 nm during 100 minutes of reaction at The XRD pattern confirmed a particle size of
room temperature. The plank tube contained approximately 74 nm, and the specific surface area
(3.3) mL of absolute ethanol and (0.5) mL of the was determined to be 19.16 m²/g. Importantly,
sample, and the control tube contained (3.3) mL the morphology index (MI) derived from the
of absolute ethanol. And (0.5 mL) of DPPH. The full width at half maximum (FWHM) of the XRD
 removal percentage was calculated according to data provided insights into the interrelationship
the equation shown below. between particle size and specific surface area.

The XRD data obtained during chemical synthesis
1 − sample absorbency of TiO₂ nanoparticles. Noteworthy observations
Antioxidant Activity = 100 − × 100
control absorbency include specific 2θ angles corresponding to

Fig. 1. Crystallographic Insights: X-ray Diffraction Pattern of Chemically

Synthesized TiO2.
 
J Nanostruct 14(4): 1057046-1057, Autumn 2024 1049
 M. Ali et al./ Synthesis of TiO2 NPs and Application for Reduction of Bacterial Growth

crystallographic planes within the TiO₂ lattice. resistance of staphylococcal species to the
The d-spacing values provide information about antibacterial nature of these TiO2 nanoparticles
interplanar distances, while the grain sizes indicate the means that these nanoparticles may become
the dimensions of individual crystallites. less resistant over time. Also, P. aeruginosa, a
common and possibly resistant bacterium, also
UV-Vis absorbance spectrum of TiO2 has an CIM value of 25 mg/ml; this, then, confirms
The UV-Vis absorbance spectrum of TiO2 shown the requirement of high TiO2 levels to ensure
in (Fig. 2) offers critical insights into its optical repression. Unlike its relative P. mirabilis which
behavior. Notably, a strong absorbance peak denotes a MIC value which is higher by an order
occurs around 385 nm, indicating the material’s of 10 animalia just as the animals. At one hundred
interaction with light. and fifty-nine DDL/ml, the bacterial cells do not
appear to be hampered by TiO2 nanoparticles as
Minimum Inhibitory Concentration (MIC) strongly as in the other experiments. Such a low
The MIC values of chemically synthesized MIC value means that just a small amount of TiO2
titanium dioxide (TiO2) nanoparticles against four nanoparticles is necessary to control the growth
different bacterial species are shown in the. Table of P. mirabilis, which may make TiO2 nanoparticles
1. an efficient and inexpensive treatment against
two bacterial isolates i.e. Staph. epidermidis infections caused by this microorganism.
and Staph. haemolyticus from the data have the
higher MIC value of 25 mg/mL implying that the Antioxidant activity
concentration of TiO2 nanoparticles must be The study involves testing various
high enough to inhibit the microbes. To intrinsic concentrations of TiO2 NPs (0.0 µg/mL, 25 µg/

Fig. 2. UV-Vis Absorbance Spectrum of Chemically Synthesized TiO2. 

 
Table 1. MIC of TiO2 NPs against some pathogenic bacteria
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1050 J Nanostruct 14(4): 1057046-1057, Autumn 2024
 M. Ali et al./ Synthesis of TiO2 NPs and Application for Reduction of Bacterial Growth

mL, 50 µg/mL, 100 µg/mL, 150 µg/mL, and 200 activity increases to 2.79%. The antioxidant activity
µg/mL) to determine their potential antioxidative sees a significant increase at 150 µg/mL and
properties. Table 2 From the data, it is evident 200 µg/mL, where it reaches 9.37% and 10.82%,
that the base absorbance without any TiO2 (0.0 respectively, accompanied by a more pronounced
µg/mL) is 0.822 with 0.0% antioxidant activity, drop in absorbance to 0.745 and 0.733.
serving as the control. As the concentration of
TiO2 NPs increases, there is a gradual decrease Anticancer activity
in absorbance and a corresponding increase in The effects of various concentrations of
antioxidant activity. For instance, at 25 µg/mL, the chemically synthesized titanium dioxide
absorbance decreases slightly to 0.810, yielding a nanoparticles (TiO2 NPs) on Human Skin Squamous
modest antioxidant activity of 1.46%. This trend Cell Carcinoma (HSSCC) over a period of 24, 48, and
continues, with the antioxidant activity mildly 72 hours. The results are displayed as mean values
increasing to 1.82% at 50 µg/mL. with corresponding standard deviations, and each
A notable shift in both absorbance and experiment was conducted with two replicates
antioxidant activity is observed at higher (N=2). Concentrations of TiO2 NPs ranged from
concentrations. At 100 µg/mL, the absorbance 0 µg/mL (control) to 1000 µg/mL. Table 3 As
further decreases to 0.799, and the antioxidant observed from the table, the control group (0

Table 2. Impact of TiO2 NPs Conc. on antioxidant activity


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Table 3. Effects of different concentrations 2
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Table 4. Effectiveness of chemically synthesized TiO2 NPs on biofilm inhibition in all bacterial
isolates.
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J Nanostruct 14(4): 1057046-1057, Autumn 2024 1051
 M. Ali et al./ Synthesis of TiO2 NPs and Application for Reduction of Bacterial Growth

µg/mL) maintained relatively stable cell viability prominently as the exposure time increased from
across all time points, with negligible changes 24 to 72 hours. Notably, the viability sharply
noted. However, increasing concentrations of decreased at higher concentrations, with the
TiO2 NPs led to a decrease in cell viability, more most substantial reductions observed at 250 µg/

Table 5. Antibacterial Activity of Chemically Synthesized TiO₂ Nanoparticles at Varying Concentrations Against Different Microorganisms
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Fig. 3. Antibacterial activity of TiO2 nanoparticles chemical method.
 


Fig. 4 .Comparison of Antibacterial Efficacy between Titanium Dioxide Nanoparticles with Antibiotics Across Different Bacterial
  Strains

1052 J Nanostruct 14(4): 1057046-1057, Autumn 2024

 M. Ali et al./ Synthesis of TiO2 NPs and Application for Reduction of Bacterial Growth

mL and 500 µg/mL, where cell viability dropped to TiO2 NPs were applied. The results demonstrate
0.76±0.01 and 0.61±0.01 at 72 hours, respectively. a distinct variance in effectiveness based
Complete cell eradication was achieved at the on the type of bacterial isolate. Both Staph.
highest concentration of 1000 µg/mL across all haemolyticus and Staph. epidermidis showed
time points. 100% biofilm inhibition when treated with the TiO2
Statistical analysis, indicated by P values, shows nanoparticles, indicating that these nanoparticles
significant differences (P < 0.05) at many points, are highly effective against the biofilms formed
particularly notable at concentrations of 7.8 µg/ by these particular staphylococcal species. Table
mL and higher. Lower concentrations typically 4 However, the effectiveness of TiO2 NPs varied
did not result in statistically significant changes significantly with other bacterial species. For P.
compared to the control. mirabilis, the biofilm inhibition was only 28.9%,
suggesting a markedly lower susceptibility of
Antibacterial activity P. mirabilis biofilms to the antibacterial effects
The biofilm inhibition by titanium dioxide of TiO2 NPs. In the case of P. aeruginosa, the
nanoparticles (TiO2 NPs) synthesized chemically, nanoparticles were moderately effective, achieving
compared against a control setting where no a 61.9% inhibition of biofilm formation. This data

Table 6. Synergism effect of Titanium Dioxide Nanoparticles and Antibiotics Against Various Bacterial Isolates
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E/d ϲϬ Ϯϰ ϭϱ Ͳϰϰ ϭϰ Ϯϱ Ͳϲϰ͘ϳϬ ϲ ϭϳ ϵϬ͘ϵϬ Ϯϭ ϭϭ
EKZ ͲϭϮ ϮϮ Ϯϱ Ͳϲϲ͘ϲϲ ϲ ϭϴ ϭϴ͘ϭϴ Ϯϲ ϮϮ ϮϱϬ Ϯϭ ϲ
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D Ϯϭϲ͘ϲϲ ϭϵ ϲ Ͳϳϭ͘ϰϮ ϲ Ϯϭ Ͳϯϱ ϭϯ ϮϬ ϭϯϯ͘ϯϯ ϭϰ ϲ
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Fig. 5. Elemental dispersive spectrum analysis.
 
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 M. Ali et al./ Synthesis of TiO2 NPs and Application for Reduction of Bacterial Growth

underscores that while TiO2 nanoparticles can be mirabilis, which is one strain against which they
highly effective against certain bacterial biofilms, exhibit substantial inhibition. On the other hand,
their efficacy can differ substantially depending depending on the bacterial strain, certain drugs
on the specific microorganism. The Antibacterial show stronger inhibition than others, indicating
capacity of TiO2 nanoparticles (NPs) synthesized inconsistent efficiency. Remarkably, when it comes
chemically at different concentrations against to preventing bacterial growth, TiO2 NPs are either
several bacteria. In their studies, they have more effective than some antibiotics or at least
demonstrated outcomes for Staph. haemolyticus, [Link] 6 and Fig. 4.
Staph. epidermidis, P. mirabilis, and P. aeruginosa.
Bacterial growth is inhibited for each bacterium Elemental Dispersive Spectrum(EDS)
and the level of inhibition is presented as zones Using EDS spectra,the elemental analysis
of inhibition in millimeters, with information on of the chemical compounds was examined.
three individual measurements and a mean with The EDS spectra of bio-mediated TiO2 NPs are
standard deviation Table 5. displayed in Fig. 5. The peaks at certain energies
A distinct trend can be observed across the on the elemental analysis graph, the height of
results: the antibacterial activities of the TiO2 which is inversely correlated to the element’s
nanoparticles might be substantially lower when concentration. Elements like carbon (C), oxygen
their concentration is decreased to 20% from (O), sodium (Na), and others are identified in the
100%. For example, S. haemolyticus has got more spectrum, with titanium (Ti) displaying the largest
than 25% reduction in zone of inhibition from weight proportion. The weight percentages and
28. 33±1. 52 mm at 40% to 14. The maximum standard deviations for every element are given
concentration of the pollutant is 66±1. 15 mm at in the box labelled “Spectrum 9,” providing a
20% concentration. Another bacterium from the quantitative examination of the elements.
Staphylococcus family, Staph. epidermidis, shows The elemental data composition and distribution
a similar decrease from 24±1 mm to 13±2 mm of titanium tetraisopropoxide (TTIP) nanoparticles
with the same concentration gradient Fig. 3. was obtained through their characterization.
The results show different degrees of inhibition, Several peaks with varied sizes and intensities
with TiO2 NPs being particularly efficient against P. were found by the analysis, suggesting that the


Fig. 6. Elemental composition and distribution of TTIP Nanoparticles based on peak intensity and area analysis
 

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 M. Ali et al./ Synthesis of TiO2 NPs and Application for Reduction of Bacterial Growth

sample included a variety of components. Peak which relates to band-gap energy changes accord
4 stood out as having the maximum intensity at with Zhang et al. (2020) who stated that similar
1035.77 and a significant corrected area of 0.595, shifts could assist in improving the light absorbing
indicating the presence of a large component. ability of the material [22]. It lost efficacy against
Peaks 10 and 12 were prominent in the sample, Staphylococcus species and P. aeruginosa at MIC
as seen by their considerable intensities and of 25 mg/mL while it showed susceptibility against
corrected areas. On the other hand, a few peaks, P. mirabilis at very lower concentration of MIC 1.
such peaks 1 and 13, showed reduced corrected 56 mg/[Link] is in line with those of [23], who
areas, suggesting small components or possible also found that different bacteria species exhibited
problems with baseline correction Fig. 6. differential resistance to nano- TiO2, possibly
caused by the differences in the cell walls and the
Scanning Electron Microscope metabolic activities. Staphylococcus species shows
The prepared TiO2 NPs’ SEM images are high resistance level. Hence more concentration of
displayed in (Fig. 7). The surface morphology the TiO₂ or combined treatment strategies may be
of chemically synthesized TiO2 nanoparticles required for better management of the microbes
resembles a sphere, while the bio-mediated TiO2 [24].
nanoparticles have a spherical structure as seen The TiO2 nanoparticles’ antioxidant activity
in the SEM picture. It was discovered that the concentration-dependent increase is remarkable
average particle size of spherically formed TiO2 NPs since it increases with the concentration,
was in the 39 nm range. resulting in a higher antioxidant capacity at higher
The X-ray diffraction results of the study concentrations. This is the same trend reported by
showed that the nanoparticles of TiO₂ were [25], who suggested that the antioxidative stress
predominantly in the anatase phase, tetragonal, activity of nanoparticles might be due to their
with a calculated particle size of 74 nm and a surface properties and ability to remove reactive
surface area of 19 m2/g. 16 m²/g. These results oxygen species. These characteristics show the
are in agreement with those concluded by Zhang possibility of using TiO2 nanoparticles as protective
and Banfield (2000) who synthesized anatase TiO₂ coatings, as well as in the form of antioxidants in
nanoparticles with the same structural features pharmaceutical formulations [26].
and highlighted the role of particles with small The statistically significant dose-dependent
sizes in increasing the photocatalytic activity due toxicity of TiO2 nanoparticles for HSSCC cell
to the increased surface area [21]. Also, the UV- line has been demonstrated, agreeing with
Vis absorbance spectrum with the blue shift and the conclusions drawn by the study by Čapek


Fig. 7. SEM images of TiO2 nanoparticles

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 M. Ali et al./ Synthesis of TiO2 NPs and Application for Reduction of Bacterial Growth

and Roušar (2021), where such effect has been UV-B radiation the means of how exactly the UV
observed. They have identified and proved the wavelengths’ type of radiation can be used on the
fact that TiO2 nanoparticles cause apoptosis via cancer treatment. Such a treatment is fair because
the entering of molecular reactive oxygen species these radiation wavelengths are going to damage
and damage of the mitochondrion’s functionality only cancer tissues.
[27]. The clearly poisonous effects arise at the
higher concentrations (250 μg/mL and 500 μg/ CONFLICT OF INTEREST
mL) signify that TiO₂ nanoparticles can be used The authors declare that there is no conflict
as therapeutic medications for cancer treatment, of intere sts regarding the publication of this
especially because they can discriminatingly then manuscript.
target and destroy cancer cells at the required
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