Ghareeb et al.

Microbial Cell Factories (2024) 23:341 Microbial Cell Factories

[Link]

REVIEW Open Access

Unlocking the potential of titanium dioxide

nanoparticles: an insight into green
synthesis, optimizations, characterizations,
and multifunctional applications
Ahmed Ghareeb1, Amr Fouda2*, Rania M. Kishk3 and Waleed M. El Kazzaz1

Abstract
This comprehensive review explores the emergence of titanium dioxide nanoparticles (TiO2-NPs) as versatile
nanomaterials, particularly exploring their biogenic synthesis methods through different biological entities
such as plants, bacteria, fungi, viruses, and algae. These biological entities provide eco-friendly, cost-effective,
biocompatible, and rapid methods for TiO2-NP synthesis to overcome the disadvantages of traditional approaches.
TiO2-NPs have distinctive properties, including high surface area, stability, UV protection, and photocatalytic activity,
which enable diverse applications. Through detailed analysis, this review demonstrates significant applications of
green fabricated TiO2-NPs in biomedicine, explicitly highlighting their antimicrobial, anticancer, and antioxidant
activities, along with applications in targeted drug delivery, photodynamic therapy, and theragnostic cancer
treatment. Additionally, the review underscores their pivotal significance in biosensors, bioimaging, and agricultural
applications such as nanopesticides and nanofertilizers. Also, this review proves valuable incorporation of TiO2-NPs
in the treatment of contaminated soil and water with various environmental contaminants such as dyes, heavy
metals, radionuclides, agricultural effluents, and pathogens. These comprehensive findings establish the foundation
for future innovations in nanotechnology, underscoring the importance of further investigating bio-based synthetic
approaches and bioactivity mechanisms to enhance their efficacy and safety across healthcare, agricultural, and
environmental applications.
Keywords TiO2-NPs, Biogenic synthesis, Biomedical applications, Targeted drug delivery, Bioimaging, Biosensors,
Agricultural applications, And environmental remediation

*Correspondence:
Amr Fouda
amr_fh83@[Link]
1
Botany and Microbiology Department, Faculty of Science, Suez Canal
University, Ismailia 41522, Egypt
2
Botany and Microbiology Department, Faculty of Science, Al-Azhar
University, Nasr City, Cairo 11884, Egypt
3
Microbiology and Immunology Department, Faculty of Medicine, Suez
Canal University, Ismailia 41522, Egypt

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 2 of 40

Introduction modulating the particle size, materials’ structural, elec-

In the 1980s, nanotechnology breakthroughs allowed tronic, physical, and chemical properties can be tailored
scientists to manipulate atomic particles, enabling the to meet specific requirements across various applica-
engineering of nanoscale materials and structures. This tions. Therefore, as a result, oxide nanoparticles, owing
innovation has applications in biomaterials, organic to their size limitations and abundant density of corner
chemistry, and medicine [1]. This microscopy technol- surface sites, possess the ability to exhibit unique physical
ogy has reshaped modern medicine, introducing new and chemical properties [16].
approaches to drug delivery, body imaging, and disease Due to the high production of titanium oxide nanopar-
detection with remarkable precision [2]. Nanotechnol- ticles (TiO2-NPs), approximately 10 × 103 tons in 2011,
ogy has progressed challenging diagnoses and advanced it is considered the major and promising industrial sub-
understanding of disease mechanisms. Due to their stance [17]. TiO2-NPs owe their widespread use to their
molecular-level size, nanoparticles enhance treatments economical production costs, superior chemical stability,
in both in-vivo and in-vitro lab settings. These micro- elevated refractive index, robust oxidation capabilities,
scopic carriers facilitated targeted drug delivery in high- and the existence of oxygen vacancies in their crystalline
risk areas, delivering exact doses while minimizing side lattice [18]. The substantial band gap exhibited by TiO2-
effects [3]. NPs is a pivotal property that renders them suitable for
Nanoparticles (NPs) possess significant advantages semiconductor applications in the optical industry [19].
compared to bulk materials, including expanded surface Attributed to the exceptional electrical and ionic prop-
area, elevated surface energy, constrained nano environ- erties of TiO2-NPS, these materials can be further cus-
ment, and decreased imperfections [4]. Characteristics tomized and tailored to be utilized in the fields of sensor
of nanoparticles, like size, shape, chemical makeup, sur- technology and electronic device fabrication [20].
face architecture, and crystallinity, heavily influence their TiO2-NPS manifests as a white, water-insoluble powder
potential uses across many areas, including pharmaco- with a remarkably high refractive index of 2.4, rendering
logical domains such as antimicrobial [5, 6], antioxidant it suitable for pigments in the paint industry [21]. Nota-
[7], anticancer [8], anti-tyrosinase potential [9] as well as bly, TiO2 naturally occurs in three distinct polymorphic
industrial and optoelectronics domains [10]. forms - rutile, anatase, and brookite - each possessing a
In this regard, transition metals have garnered sig- crystalline structure. These polymorphs find extensive
nificant attention due to two key factors: firstly, their applications in the gemstone industry [22]. Furthermore,
atomic electronic configuration features incomplete owing to their unique crystalline nature, the physical
d-orbitals, and secondly, their ability to undergo fluc- and chemical properties of TiO2-NPs can be tailored by
tuating oxidation phases. Consequently, their intrinsic modifying the ratio of these polymorphic forms, thereby
physicochemical properties can be tailored for a vast expanding their scope of applications across various
array of applications across the fields of natural sci- sectors. The remarkable versatility of TiO2-NPs is high-
ences and materials engineering [11]. Furthermore, lighted by their extensive and successful applications
these metallic elements can generate a diverse range of across diverse domains. These nanoparticles have proven
oxide compounds, which present promising prospects invaluable in the field of sensors [23]. While also serv-
for expanding the frontiers of research across burgeon- ing as photocatalysts for the decomposition of wastewa-
ing domains such as environmental studies, agricultural ter pollutants [24]. Furthermore, their antimicrobial and
applications, medical advancements, cosmetic formula- antibacterial properties [25] allow for their use as food
tions, energy storage solutions, fuel cell technologies, additives and in cosmetic products [26].
semiconductor device development, sensor innova- In this review, we delve into different eco-friendly, safe,
tions, and catalytic processes [12]. Particle size is a piv- and sustainable synthesis techniques for TiO2-NPs pro-
otal factor that significantly influences the fundamental duction. This eco-friendly approach offers significant
properties of any material. Firstly, with regard to struc- advantages over conventional industrial methods, nota-
tural properties, bulk oxides exhibit robustness and low bly improved reagent handling and enhanced process
phase stability, whereas nanoparticle oxides can readily safety [27]. Furthermore, to the best of our knowledge,
undergo structural transformations and possess low sur- we have endeavored to provide a comprehensive sum-
face free energy due to their reduced particle size [13]. mary of the various biomedical applications of TiO2-NPs,
Secondly, concerning electronic properties, the surfaces encompassing their antioxidant, antibacterial, antifungal,
of bulk oxides are extended due to the distribution of antiviral, and anticancer with other biotechnological and
ionic charge as compared to the nanoscale structure [14]. environmental applications. This review aims to elucidate
Thirdly, the band gap of bulk oxides is broad. In con- our current understanding within this field and inspire
trast, NP oxides exhibit a low band gap, enhancing con- the exploration of more sophisticated nanostructures
ductivity and chemical reactivity [15]. Consequently, by in the forthcoming years. By elucidating the synthesis

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 3 of 40

methods and potential applications, we strive to pave routes for nanoparticle synthesis, potentially enabling
the way for future advancements and innovations in the unique geometric configurations.
realm of TiO2-NPs. Nonetheless, the methods mentioned earlier frequently
necessitate multiple processing steps, stringent control
Synthesis methods over variables such as pressure, pH, and temperature,
Conventional methods and the employment of expensive equipment and toxic
Conventional methods for synthesizing metal oxides chemicals. Furthermore, these techniques often result
can be broadly categorized into two distinct approaches: in the generation of toxic by-products that pose a sig-
top-down and bottom-up. The top-down approach nificant threat to ecosystems [34]. In light of these chal-
involves breaking down bulk macroscopic particles into lenges, there is an urgent need to develop eco-friendly
nanoscopic particles (1 mm-xµ) through various physi- approaches utilizing biological and green synthesis
cal methods. This approach is relatively straightforward techniques.
than the bottom-up but also encounters monodispersed
challenges and limited particle control. Various physical Biogenic synthesis
techniques involved, such as milling, etching, sputtering, Considering a broader perspective, biological approaches
pulse laser ablation, and evaporation-condensation tech- embrace the concept of “green synthesis” for the pro-
niques [28]. duction of nanoparticles [35]. This method can be
On the other hand, the bottom-up approach relies on a further categorized into two distinct pathways: (a) phy-
self-assembly process, whereby atomic nuclei are joined tosynthesis, where the synthesis process is facilitated by
together to form nanosized particles (0.1 nm-xµ). This leveraging the capabilities of plants and their extracted
approach facilitates the easy manipulation of nanopar- compounds, and (b) microbial synthesis, which involves
ticle dimensions and morphology, resulting in superior harnessing the synthetic potential of microorganisms
homogeneity. However, its drawbacks include scalability, such as bacteria, algae, fungi, yeasts, and actinomycetes,
strict resource constraints, and multi-phase implementa- utilizing extracts derived from these organisms. The phy-
tion. It encompasses several techniques and processes, tosynthetic route exploits the natural ability of plants to
including chemical vapor deposition, sol-gel processes, synthesize materials by harnessing their varied metabo-
hydrothermal methods, sonochemical techniques, flame lites. At the same time, the microbial pathway capitalizes
spraying, spinning, and green synthesis methods [29]. on the metabolic machinery of various microbial species
Notably, while the top-down approach focuses on reduc- to generate the desired metal and its oxides-nanoparti-
ing the size of bulk materials to the nanoscale, the bot- cles. These biological methods offer an environmentally
tom-up approach builds nanoparticles from the ground friendly and sustainable alternative to conventional syn-
up, starting with atomic or molecular precursors [30]. thesis techniques [36, 37].
The choice between these approaches depends on sev- Green synthesis surpasses conventional processes by
eral factors, such as the desired material properties, cost- integrating non-toxic biological organisms, natural acids,
effectiveness, and scalability requirements. and water as solvents and safer catalysts while replacing
Researchers have developed a wide range of chemi- hazardous chemical substances. This eco-green strategy
cal and physical approaches to synthesize nanoparticles significantly enhances reaction efficiency with faster syn-
with diverse geometries, enabling their application across thesis times and higher yields for industrial scalability
numerous fields. Among the novel techniques employed (Table 1). Concurrently, it generates substantial product
for achieving these distinct nanoparticle geometries, yields of biocompatible nanoparticles that are imperative
a clear distinction can be drawn between lithography- for use within the medical and pharmaceutical domains
based techniques and non-lithographic methods [31]. [38].
The lithography techniques, which include photolithog- The green synthesis approach faces several key chal-
raphy, ion beam lithography, microcontact printing, dip lenges that can potentially obstruct its successful imple-
pen lithography, and nanoimprint lithography, involve mentation. Firstly, the optimization processes required to
patterning materials on a surface through various litho- synthesize nanoparticles with specific size distributions
graphic processes [32]. These techniques allow for precise and morphological characteristics are inherently tied to
control over the geometric features of the nanoparticles. their intended biological functions (Table 1). Secondly,
On the other hand, non-lithographic techniques encom- deciphering the distinct role played by each constituent
pass methods such as ball milling, a mechanical process compound involved in the biofabrication process man-
for grinding and mixing materials, evaporation-con- dates a comprehensive chemical analysis of the filtered
densation, and electrochemical synthesis, which utilizes biological biomass [39].
electrochemical reactions to synthesize nanoparticles
[33]. These non-lithographic approaches offer alternative

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 4 of 40

Table 1 Summarizing the advantages and disadvantages of biogenic NPs synthesis

Biogenic NPs Advantages Disadvantages Ref-
synthesis er-
ence
Plant-Derived - Bio-safe, inexpensive, and quick Production process - Genetically unmodifiable, unlike [35,
NPs - Eco-sustainable approach (contaminants and pollutants recycling) microorganisms 36,
- Optimize synthesis oversight with stable and uniform NPs yield 38,
- Biomass waste utilization and reusability 40]
Microbial Me- - Decreased requirement for extra capping or stabilizing agents. - Scaling up manufacturing for commer-
diated NPs - Can be genetically altered for higher yields. cial production (contamination, sterility,
- Less handling of toxic chemicals & lower infrastructure requirements and cost concern).
- Quick and easy to adjust and manipulate synthesis parameters (PH, [Link].) - Multiple separation steps are needed,
- Adaptive capabilities with different synthesis routes (intracellular, extracellular, as well as batch-to-batch yield variation
and cell-free extract synthesis. with the possibility of NPs aggregation
- Cost and energy-efficient approach over time
- Versatile biomedical and industrial applications - Bioproduct quality variance.
- Constrained dimensional control and
NPs instability for particular NPs

Phytosynthetic routes producing various biogenic nanoparticles with important

The synthesis of nanoparticles facilitated by plants is antimicrobial applications [48].
considered more stable compared to the synthesis medi- The desired plant part undergoes thorough washing
ated by microbes. The process of producing nanoparti- and cleaning, followed by boiling in a solvent like ethanol
cles using plant extracts is straightforward, economical, (C2H6O) or dH2O and subsequent filtration (Fig. 1). The
and yields a high quantity while adhering to sustain- resultant filtered solution, rich in plant extracts, serves
able practices [41]. These plant extracts can be derived as a reducing agent. To this filtered solution, a suitable
from various plant parts, such as flowers, roots, seeds, metallic or metallic oxide precursor such as titanium tet-
or leaves. Among the different plant parts, leaves are raisopropoxide (Ti(OCH(CH3)2)4), titanium tetrachloride
more commonly utilized for obtaining extracts, as they (TiCl4), or titanyl hydroxide (TiO(OH)2) for TiO2-NPs
are abundant in metabolites [42]. Leaves offer a more synthesis is introduced under constant agitation [49]. The
viable option for deriving extracts without generating reaction mixture starts vigorously when the TiO2 precur-
toxic byproducts. These processes involving leaf extracts sor salt is combined with the plant extract, and a color
are straightforward, economical, and non-toxic, making change indicates the first sign of nanoparticle synthesis
them easily accessible and compatible for NPs produc- [50]. This initial observation can then be confirmed after-
tion with appropriate quantities suitable for industrial ward by spectroscopic techniques, leading to the forma-
and environmental sectors [43]. tion and characterization of the desired TiO2-Nps [51].
Significantly, plant extracts are abundant in poly- A major portion of green synthesis studies utilize leaf
phenolic compounds that function as potent reducing extracts, as leaves are abundant reservoirs of metabolites.
agents, thereby facilitating the reduction of metal ions For example, Extracts from Moringa oleifera leaves facili-
and consequently leading to the formation of nanopar- tated the synthesis of TiO2-NPs around 100 nm in size
ticles [44]. Additionally, the reduction of these metallic with varying shapes, exhibiting promising wound healing
ions can be prompted by various cellular components capabilities [52]. Another TiO2-NP was synthesized using
such as amines, carbonyls, phenolic compounds, pig- the leaf extract of Hibiscus rosasinensis L. and exhibited
ments, terpenoids, and alkaloids [45]. This synthesis pro- potent antimicrobial activity against both Gram-nega-
cess is driven by redox reactions involving metallic ions tive and Gram-positive bacterial strains [53]. In another
and secreted molecules like sugars, carbohydrates, and study, TiO2-NPs derived from Nyctanthes leaf extracts
proteins. However, elucidating the precise mechanism possessed a uniform spherical shape and ranged in size
of action remains challenging due to the vast chemical from approximately 100 to 150 nm. These biogenic NPs
diversity of the metabolites involved in the reduction demonstrated significant pediculicidal (lice-killing), acar-
process [46]. Nevertheless, experimental parameters icidal (mite-killing), and larvicidal (larva-killing) proper-
such as pH, temperature, reactant concentration, and ties [54]. Similarly, the leaf extract of Calotropis gigantea
reaction time play a pivotal role in determining the phys- L. was reported to facilitate the reduction of TiO2 to
ical-chemical properties of the resultant nanoparticles nanoparticles within a span of 6 h, attributed to the pres-
[47]. Moreover, the biodiversity and availability of differ- ence of primary amines in the extract. The bio-mediated
ent plant families, each with unique profiles of primary TiO2-NPs exhibited promising acaricidal activity against
and secondary metabolites, contribute significantly to

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 5 of 40

Fig. 1 Flowchart showing plant-mediated production of TiO2-NPs starting from plant extract to applications

the larvae of Rhipicephalus microplus and Haemaphysa- geometric parameters and form of the resultant nanopar-
lis bispinosa [55]. ticles [90].
The synthesis of TiO2-NPs has also been reported using The formation of nanoparticles can occur through
the aqueous leaf extract of the medicinal plant Cathar- either intracellular or extracellular mechanisms, contin-
anthus roseus (L.). The aliphatic alcohols and amines gent upon the specific type of microorganism involved
present in the extract contributed to the formation of [91]. Researchers have leveraged the potential of living
TiO2-NPs with irregular morphologies and particle cell extracts to pursue biological nanoparticle synthesis.
sizes ranging between 25 and 110 nm [56]. Notably, the The subsequent sections will concisely explore the pri-
variation in size and morphology observed among the mary biological pathways employed for this synthesis.
reported studies is influenced by factors such as reaction
temperature, time, and the source of the plant extract Bacterial-mediated TiO2-NPs biosynthesis Bacteria
[57]. Thus, optimizing these parameters can lead to are favored for nanoparticle synthesis due to their rela-
improvements in the synthesis mechanism [58]. Further- tively undemanding conditions, uncomplicated purifica-
more, a wide array of propitious plant extracts remains to tion processes, and prolific generation. Consequently,
be explored for synthesizing TiO2-NPs. Table 2 shows the these microorganisms have emerged as extensively stud-
green synthesis of TiO2-NPs using plant extracts regard- ied " nanomaterial production hubs“ [36]. Depending on
ing their sizes, shapes, and applications. their specific characteristics, various bacterial species
can synthesize these NPs intracellularly or extracellularly
Microbial synthesis (Table 3). The bacterial synthesis of TiO2-NPs is aided
The green synthesis of NPs follows a bottom-up strat- by their natural defense mechanisms, allowing them to
egy, where NPs form through the oxidation/reduction adapt to environmental changes and resist certain metals.
of metallic ions facilitated by biomolecules secreted by This process can occur through three primary methods:
microorganism entities. These biomolecules, including using whole cells, supernatants, or extracts. Superna-
enzymes, proteins, sugars, carbohydrates, and others, tants, obtained after centrifuging bacterial cultures, con-
play a crucial role in driving the redox processes that lead tain various bioactive compounds such as enzymes, pro-
to the generation of nanoparticles [88]. Despite extensive teins, amino acids, polysaccharides, and carbohydrates,
research, a comprehensive understanding of the nanopar- which are utilized as biocatalysts for NPs production [92].
ticle synthesis mechanism driven by microorganisms This bacterial-mediated approach offers a versatile and
remains elusive as each type of microorganism interacts efficient method, capitalizing on these microorganisms’
with metallic ions through various pathways [89]. The inherent properties and adaptability.
biochemical processes and interaction dynamics of a The biosynthesis of TiO2-NPs from Ti3+ ions (titanium
specific microorganism, coupled with environmental fac- precursors) by bacteria occurs through a three-step pro-
tors such as temperature and pH, ultimately influence the cess: trapping, bioreduction, and capping. Initially, bac-
teria in the aqueous solution or surrounding medium

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 6 of 40

Table 2 Examples of some plants-derived TiO2-NPs and their applications

Plant Species Size Shape Applications / Properties Studied Reference
Morus nigra (Mulberry) 0.17 nm round shape- irregular Antimicrobial activity against Staphylococcus aureus and Proteus [59]
(TiO2(Aq)), surface morphologies mirabilis
0.076 nm
(TiO2(Et))
Carthamus tinctorius 47 nm Spherical, irregular Alternative fuel additives for diesel-biodiesel blends, impact on [60]
(Safflower) engine performance and emissions
Phyllanthus niruri Industrial wastewater treatment [61]
Ivy leaf extract 26.34 nm irregular spherical and Cefoperazone removal from pharmaceutical wastewater, antimi- [62]
tetragonal shape crobial activity
Punica granatum 100–150 nm spherical Antibacterial activity against E. coli [63]
(Pomegranate)
Mentha arvensis 80.51–200 nm Triangular aggregation Antioxidant activity (DPPH, FRAP assays) [64]
pattern
Mangifera indica 3–27 nm agglomerated spheri- Antimicrobial activity against fungi and bacteria, antioxidant activ- [65]
(Mango) 8–28 nm cal-like morphology ity, non-linear optical activity
Azadirachta indica less
(Neem) agglomerated- spherical
Avicennia marina 25–35 nm irregular, rough, and Methylene Blue Removal, nitrate, and chemical oxygen Demand [66]
(Mangrove) granular surface texture (COD) reduction
Cissus rotundifolia 1–100 nm Spherical Anticarcinogenic activity against Streptococcus mutans and [67]
Lactobacillus sp.
Caesalpinia pulcherrima 20–30 nm Spherical Photocatalytic dye degradation of methylene blue, antimicrobial [68]
activity against bacteria
Mucuna pruriens 5 μm to 500 nm Triangular aggregation Antioxidant activity (DPPH, FRAP assays) [69]
pattern
Ocimum tenuiflorum 10.2–15.2 nm knitted ball-like crystal- Enhancement of growth, physiology, enzymatic activities, essen- [70]
(Holy basil) line structure. tial oil content and yield
Syzygium cumini (Java 12–30 Round and Nematicidal activity against root-knot nematode (Meloidogyne [71]
plum) polydisperse incognita), impact on plant growth and physiology
Morus alba (Mulberry) 28.34 nm Anatase crystalline Antimicrobial activity, UV protection, colorfastness to washing/ [72]
phase rubbing/light
Juniperus phoenicea 10–30 nm Spherical Insecticidal activity against Sitophilus oryzae, antimicrobial activity, [73]
impact on the nutritional value of grains
Terminalia bellirica 420 nm Spherical Antioxidant activity, antimicrobial activity [74]
Psidium guajava (Guava) 5–10 nm Spherical Antimicrobial activity against E. coli and S. aureus [75]
Salacia reticulata 32–46 nm flake-like structures Antidiabetic, anti-inflammatory, antibacterial activities, develop- [76]
mental toxicity in zebrafish embryos
Psidium guajava (Guava) 10–30 irregular Antimicrobial activity, anticancer activity against MG-63 cells, [77]
structural and optical characterization
Commelina benghalensis 150–200 nm Roughly spherical Photodegradation of methylene blue dye and sulfisoxazole [78]
antibiotic
Moringa oleifera 10 nm Anatase form Impact on germination and growth of spinach seeds [79]
(Moringa)
Ipomoea carnea (Morn- 7–10 nm. spherical Effect on growth, yield, and quality of black carrot [80]
ing glory)
Tinospora cordifolia 18–20 nm Spheroidal Photocatalytic degradation of methylene blue dye, [81]
Tulbhagia violacea 31–42 nm Rectangular Antioxidant activity, anticancer activity against HEK 293 and HeLa [82]
cell lines,
Trianthema 30–50 nm spherical Antifungal activity against Ustilago tritici (wheat fungal disease), [83]
portulacastrum 40–60 nm round
Chenopodium quinoa
Mango leaf extract 10–30 Spherical (TN-2, TN-3), Antifungal activity against Penicillium steckii [84]
agglomerates (TN-1)
Terminalia catappa 75.24 ± 1.12 nm Monoclinic-shaped, Plant modulating ability on Abelmoschus esculentus, antioxidant [85]
clustered activity, impact on plant growth and physiology
Juniperus phoenicea 10–30 nm Spherical Antimicrobial activity against various bacterial and fungal strains, [86]
anticancer activity against human ovarian adenocarcinoma cells
Moringa oleifera leaf 25–110 nm spherical Salinity Stress mitigation in wheat crops [87]

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 7 of 40

Table 3 Bacterial and actinomycetes species-mediated biosynthesis of TiO2-NPs showing promising biomedical and biotechnological
applications
Source Size Shape Applications/Properties Studied Reference
TiO2-NPs fabricated by bacterial strains
Staphylococcus aureus (G + ve) ~ 20 nm (average Smooth and Antibacterial and antibiofilm against Bacillus [112]
diameter) spherical subtilis and Escherichia coli
Lactobacillus rhamnosus (G + ve) 3–7 nm Spherical Antifungal against isolated fungal strains, Bio- [113]
compatible towards WI38 and HFB4 cell lines
Bacillus subtilis MTCC 8322 (G + ve) 80–120 nm Spherical to Photocatalytic dye degradation (Methylene Blue [114]
irregular and Orange G)
Bacillus subtilis (G + ve) 70.17 nm spherical Improved mechanical properties of Glass Iono- [115]
mer Cement (GIC) for dental applications
Paenibacillus sp. HD1PAH and Cyperus 17.11 nm Spherical Anthracene biodegradation, soil enzyme activi- [116]
brevifolius. 29.39 nm Granular ties increased
Halomonas elongata IBRC-M 10,214 (G-ve) 104.63 ± 27.75 nm Spherical Antibacterial against E. coli and S. aureus [117]
Lactobacillus johnsonii (G +ve) 4–9 nm Irregular Desalinization and surface cleaning [118]
Bacillus amyloliquefaciens (G +ve) 22.11–97.28 nm Spherical Photocatalytic dye degradation (Reactive Red [119]
31), enhanced by doping
Aeromonas hydrophila (G-ve) 40.50 nm Smooth, spheri- Antibacterial against S. aureus and S. pyogenes [120]
cal, uneven
Propionibacterium jensenii (KC545833) (G +ve) < 80 nm uniform size and Collagen stabilization for wound dressing [121]
anatase form
Bacillus subtilis (FJ460362) (G +ve) 10–30 nm Mostly spherical Photocatalytic control of aquatic biofilm [122]
TiO2-NPs formed by actinomycetes strains
Streptomyces sp. HC1 30–70 nm Spherical Antimicrobial activity, antibiofilm activity [110]
Streptomyces bluensis 58.3 nm spherical Azo dye degradation [123]
Saccharopolyspora spinosa 23.3 nm Spherical Antimicrobial [124]

trap Ti3+ ions. Subsequently, enzymes and proteins facili- bacterial colonization [100, 101]. In biogenic nanoparti-
tate the reduction of these trapped ions into TiO2-NPs. cles, EPSs exhibit intriguing characteristics. One notable
Research has shown that microbial proteins containing property is mucoadhesion, which facilitates neutral coat-
functional groups such as –NH2, –SH, –COOH, and ing with low surface energy and prevents the recogni-
–OH plays a crucial role in stabilizing the synthesized tion of non-specific protein receptors. Moreover, EPSs
TiO2-NPs [93]. These groups provide binding sites for can adsorb metal cations onto their surface, a trait that
Ti3+ ions and act as capping or stabilizing agents. The aids in stabilizing and biosynthesizing metal nanopar-
reduction of Ti3+ ions to NPs occurs either on the cell ticles. Their robust reducing and stabilizing capabilities
wall or within the bacterial intramembrane space. This make EPSs an attractive alternative biogenic substrate
process involves electron transfer from reduced com- for synthesizing nanoparticles [102]. Upon interaction
pounds to inorganic compounds, promoting bacterial between metal ions and EPSs containing reducing sug-
bioreduction of NPs. Finally, various bacterial biomol- ars, the metal ions undergo chelation, followed by their
ecules cap the reduced TiO2-NPs as natural stabilizing reduction and stabilization facilitated by various func-
agents. This capping step is essential for maintaining NP tional moieties. Prominently, polyanionic groups, chiefly
stability, a critical factor in their synthesis and application -COOH and -OPO32−, are well-documented to partake
[94]. in reducing and stabilizing metal nanoparticles. Fur-
thermore, electrostatic interactions between metal cat-
Ti3+ (titanium salts) → Bacteria-Ti3+ ( Trapping).
ionic species and anionic EPS groups like -COOH and
Bacteria-Ti3+ + e−→ TiO2​-NPs (Bioreduction)
-OPO32− have been cited as advantageous for nanopar-
TiO2​-NPs + microbial functional groups (–NH2, –
ticle synthesis. Among these functional entities, -OH,
SH, –COOH, –OH) → Capped TiO2​-NPs (Capping).
-COOH, -OPO32−, hemiacetal, and amino terminals have
been proposed to reduce metal ions from precursor salts,
Among bacterial metabolites, exopolysaccharides (EPSs), yielding the corresponding nanoparticles. Notably, -OH
which are heterogeneous organic biopolymers [95, 96] groups coordinate with metal ions, whereas their oxi-
secreted into the extracellular environment [97] and dation to form carbonyl and -COOH groups is pivotal
have a vital function in providing protective microen- during the reduction process and nanoparticle forma-
vironments for bacteria [98, 99], enhancing their toler- tion [103]. Moreover, the abundance of -OH and hemi-
ance towards biotic and abiotic stresses while facilitating acetal end groups in EPSs facilitates the reduction and

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 8 of 40

stabilization of nanoparticles, rendering them more suit- coupled with their ease of handling in laboratory settings
able for diverse applications. Additionally, monosaccha- [125]. The utilization of fungi for synthesizing metal-
rides like glucose, galactose, mannose, and fructose have lic nanoparticles has garnered substantial attention due
been mentioned to reduce metal ions, contributing to the to their possessing certain advantageous traits surpass-
mechanisms involved in nanoparticle synthesis observed ing other organisms. The ability to readily scale up and
with other EPSs [104]. streamline downstream processes, the economic viability,
Also, Actinomycetes have the ability to produce a and the presence of mycelia with an augmented surface
diverse array of secondary metabolites, have emerged as area constitute valuable advantages that warrant due con-
promising sources for the biosynthesis of nanoparticles sideration [126]. Furthermore, fungi have gained height-
with desirable surface characteristics and size control ened interest owing to their involvement in the biological
[105]. These microorganisms possess the remarkable synthesis of metallic nanomaterials, facilitated by their
capability to facilitate the production of metallic and exceptional tolerance and remarkable ability to bioaccu-
metal oxide NPs through either intracellular or extra- mulate metals [127] (Fig. 2).
cellular methodologies [106] (Table 3). The extracellular The synthesis of nanoparticles employing fungi and
approach has garnered significant commercial interest their biotechnological applications, particularly in
due to its advantage of minimizing polydispersity, a cru- medicine, are encompassed within the realm of myco-
cial factor in nanoparticle synthesis [107]. Notably, acti- nanotechnology. This scientific term, representing the
nomycetes are a class of high-content guanine and convergence of “mycology” and “nanotechnology,” holds
cytosine microorganisms primarily exploited for their significant potential due to the vast diversity and range
antibiotic production capabilities [108]. However, of fungal species [128]. Fungi can form nanoparticles
their versatility extends beyond antibiotics, as several in diverse structures spanning mesoscale to nanoscale
researchers have successfully synthesized TiO2-NPs by dimensions via enzymatic reduction, either extracellu-
harnessing the metabolic pathways of these remarkable larly or intracellularly, and through biomimetic miner-
microorganisms [109]. This further highlights the poten- alization processes. Their extensive scalability has led to
tial of actinomycetes as a valuable resource for the green a distinct preference for their utilization in nanoparticle
synthesis of nanoparticles with diverse applications. synthesis, exemplified by techniques such as thin solid
Among the recent examples, one study reported the substrate fermentation. Owing to the remarkable secre-
synthesis of spherical-shaped titanium TiO2-NPs, rang- tory capabilities of fungi, producing abundant quanti-
ing in size from 30 to 70 nm, using Streptomyces sp. HCl. ties of extracellular enzymes or proteins becomes viable
The developed TiO2-NPs were tested as antimicrobials [129]. Furthermore, the economic feasibility and sustain-
against various pathogenic microorganisms, including ability of employing biomass present another advantage
Staphylococcus aureus, Escherichia coli, Candida albi- for the implementation of a green approach, enabled
cans, and Aspergillus niger. The researchers concluded by fungal entities or byproducts, in the fabrication of
that the synthesized TiO2-NPs exhibited higher antimi- metallic nanomaterials [130]. Here are some examples of
crobial properties against bacterial species compared to different fungal species used for the biosynthesis of TiO2-
fungal species [110]. Another study reported the synthe- NPs, referring to their activities (Table 4).
sis of TiO2-NPs using the marine actinobacteria Strep- Yeasts, a class of fungi belonging to the ascomycetes
tomyces bluensis as the biological source. The precursor group, have demonstrated remarkable potential for the
for the nanoparticle synthesis was Ti(OH)4. The resulting synthesis of nanoparticles. Biological processes possess a
spherical TiO2-NPs exhibited an average size of 37.54 nm superior ability to control the morphology of materials.
and exhibited high potential to degrade azo dyes, includ- During the logarithmic growth phase, the yeast Schizo-
ing AR-79 and AR-80, with percentages of 84% and 85%, saccharomyces pombe facilitated the synthesis of semi-
respectively [111]. Table 3 shows some bacterial and conductor nanocrystals [126]. Nanoparticles synthesized
actinomycetes species that were utilized for TiO2-NP extracellularly by yeasts offer a wide range of advantages,
synthesis. as the protein-mediated interactions between the yeasts
and nanoparticles facilitate subsequent downstream
Fungal-mediated TiO2-NPs biosynthesis Fungi have reactions and modifications [131]. The entire yeast family
gained widespread nanoparticle biosynthesis adoption exhibits the remarkable capability of aggregating various
due to their metabolites’ remarkable efficacy in fabri- heavy metals. They can sequester substantial amounts of
cating diverse nanoparticles. They represent a valuable toxic metal ions. Numerous studies have concluded that
addition to the repertoire of microorganisms employed the mechanisms employed by these species to counter-
for nanoparticle production. The extensive utilization of act the toxic effects of heavy metals involve the extracel-
various fungal species can be attributed to their capacity lular production of polysaccharides or peptides. These
to secrete substantial quantities of proteins or enzymes, biomolecules play a crucial role in either managing the

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 9 of 40

Fig. 2 Promising characteristics of fungi as a green tool for NPs synthesis

cell’s permeability barrier against heavy metals or actively biosynthesis of TiO2-NPs using yeast strains were sum-
effluxing them from the cell [132]. For direct intracellu- marized in (Table 4).
lar delivery of metal ions, yeast cells need to be disrupted
to avoid negative or lethal consequences. The toxicity to Viruses-mediated TiO2-NPs biosynthesis Viruses pro-
cells can be mitigated by either maximizing the retention vide an important illustration of how nanosized particles
of metal ions within the cells or by exposing them to met- can be synthesized. So far, viruses have produced nano-
als that do not possess the same level of toxicity as lead, tubes and nanorods [148]. Research studies and experi-
mercury, and cadmium ions [133]. ments have demonstrated that plant viruses and certain
The significant variations observed in size, particle dis- bacteriophages can be readily isolated and subjected to
tribution, monodispersity, and characteristics of the syn- further processing [149]. However, not all virus compo-
thesized nanoparticles can be attributed to the diverse nents can synthesize nanoparticles, and the underlying
mechanisms employed by different yeast strains from reasons for this need to be thoroughly investigated and
various families for nanoparticle formation. In yeast understood [150]. One study reported the successful for-
cells, a detoxification mechanism involving glutathione mation of chalcogenide nanocrystals within genetically
(GSH) and two classes of metal-binding proteins, namely modified virus-like particles [151]. Leveraging the self-
metallothioneins and phytochelatins (PC), is utilized to assembly capabilities of the genetically modifiable M13
facilitate the formation of nanoparticles [89]. In most virus, TiO2 nanostructures with controlled sizes between
yeast species, these molecules (GSH, metallothioneins, 20 and 40 nm were synthesized under ambient condi-
and PC) facilitated the formation pathway of nanopar- tions, enabling their homogeneous distribution and per-
ticles and stabilized the resulting compounds. The resis- colated networks suitable for exploiting their promising
tance mechanism was defined as the ability of yeast photo-electrochemical properties [152] (Table 5).
cells to convert the absorbed metal ions into non-toxic
polymer complexes. The nanoparticles formed by yeasts Algae-mediated fabrication of TiO2-NPs Algae are
are commonly referred to as “semiconductor nanocrys- capable of accumulating significant quantities of heavy
tals” or “quantum semiconductor nanocrystals”. Recent metals, an attribute that enables them to be utilized for the
research has revealed that yeasts can also synthesize vari- biosynthesis of metallic and metal oxide NPs [153]. Algae
ous other types of nanoparticles [126]. More examples of have found widespread application in the synthesis of

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 10 of 40

Table 4 Examples of biosynthesis of TiO2-NPs using different fungal species with reference to their size, shape, and activities
Species Size Shape Applications / Properties Studied Reference
TiO2-NPs produced by multicellular fungi
Aspergillus eucalypticola SLF1 33 nm Mesoporous, anatase Photocatalytic, antimicrobial, antioxidant activity [134]
phase
Paraconiothyrium brasiliense 57.39 ± 13.65 nm Spherical Antibacterial, antibiofilm properties [135]
Fomitopsis pinicola 80–120 nm Irregular with a rough Antibacterial and anticancer activity [136]
surface
Trichoderma viride 10.4 to 45.8 nm Elliptical to spherical Larvicidal, antifeedant, pupicidal activity against Helicoverpa [137]
(size range), armigera
26.619 ± 7.577 nm
(mean size)
Trichoderma citrinoviride 10–400 nm Irregular, triangular, Antibacterial activity against Pseudomonas aeruginosa, [138]
pentagonal, spherical, antioxidant potential
rod-shaped
Trichoderma harzianum 50.0 nm Spherical Enhanced growth of T. harzianum, inhibitory against Sclero- [139]
tinia sclerotiorum
Aspergillus niger DS22 10.4 to 45.8 nm, Elliptical to spherical Antibacterial, anti-inflammatory, wound-healing activities [140]
26.619 ± 7.577 nm
(mean size)
Aspergillus flavus 62–74 nm Spherical, oval Antibacterial activity [141]
Metarhizium anisopliae 9.50 nm Spherical Larvicidal, pupicidal, antifeedant activity against Spodoptera [142]
frugiperda
Hypsizygus ulmarius ~ 80 nm Spherical Antibacterial and anticancer potential [143]
Pleurotus djamor 31 nm Spherical Mosquito larvicidal, antibacterial, anticancer effects [144]
Streptomyces sp. HC1 30 to 70 nm Spherical Antimicrobial and antibiofilm activity [110]
Alternaria solani 15 nm Agglomerated Antimicrobial, anticoagulant, antiplatelet, hemolytic and [145]
cytotoxicity properties
TiO2-NPs produced by yeasts
Saccharomyces cerevisiae 8–35 nm Individual aggregate Photocatalytic [146]
(Baker’s yeast) antimicrobial activity
Saccharomyces cerevisiae 6.7 ± 2.2 nm Spherical Photocatalytic activity, antimicrobial activity [147]

Table 5 Examples of the biosynthesis of TiO2-NPs using viruses and algae with noted size, shape, and applications
Source Size Shape Activity Reference
Viruses
M13 bacteriophage 20–40 nm quasi-spherical photo-electrochemical properties [152]
Algae
Bostrychia tenella (Red macroalga) 22.86 nm Uniform, monodispersed Antifouling activity [160]
Carteriospongia foliascens (Marine sponge) 8.3 nm uniform, monodispersed Antifouling activity
particles
Sargassum wightii (Brown macroalga) 20–80 spherical Larvicidal activity against mosquito [161]
malaria’s vector
Phaeodactylum tricornutum (Microalga) 50 nm spherical Antimicrobial activity, antistatic properties, [162]
cytotoxicity against cancer cell lines
Sargassum myriocystum (Brown macroalga) 50–90 nm Cubic, square, spherical larvicidal activity against mosquito larvae, [163]
photocatalytic dye degradation
Spirulina platensis (Microalga) 4.62 nm Spherical, dispersed Antifungal Activity [164]
irregularly
Spirulina platensis 90–150 nm spherical Antimicrobial activity [157]

TiO2-NPs due to their ready availability and effectiveness. to bacterial synthesis routes [154]. Additionally, algae
Beyond just enzymes and proteins, these phytosynthetic can synthesize nanoparticles utilizing their extracts or
organisms also possess carotenoids and various pigments supernatants, which are rich in secondary metabolites.
involved in photosynthesis, which contribute significantly These extracts obviate the need for live algal cultures in
to the physio-assisted (algae-mediated) production of nanoparticle synthesis [155].
TiO2-NPs. However, algal-based synthesis methods for In one study, TiO2-NPs were synthesized using the
these nanoparticles are not as well-established compared algae Spirulina platensis [156]. Moreover, another study

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 11 of 40

reported the synthesis of spherical-shaped TiO2-NPs Effect of Precursor and reducing Agent concentrations on
ranging from 90 to 150 nm in size by utilizing an extract nanoparticle size and agglomeration
from S. platensis [157]. Also, TiO2-NPs were synthesized Precursor and reducing agent concentrations crucially
from the seaweed Sargassum wightii, and their efficacy determine the synthesized nanoparticles’ size [168]. Ini-
was evaluated for killing the larvae of vectors that trans- tially, excessive reducing agents bind to preformed nuclei,
mit malaria and filariasis [158]. Another investigation amplifying secondary ion reduction on their surfaces.
involved the synthesis of negatively charged TiO2-NPs This accelerates nanoparticle growth, yielding larger sizes
with cubic, square, and spherical shapes ranging approxi- at higher reactant concentrations [169]. However, an
mately 50–90 nanometers in size, utilizing Sargassum overly high reducing agent concentration adversely pro-
myriocystum as the source material. Additionally, the motes nanoparticle bridging and aggregation. This arises
researchers in this study assessed the antimicrobial prop- when an abundance of metal ions adsorbs onto nuclei
erties of the synthesized titanium dioxide nanoparticles surfaces, facilitating uncontrolled secondary reduction
[159]. Table 5 summarizes some examples of the biosyn- and agglomerated growth [170]. Therefore, striking the
thesis of TiO2-NPs using different algal strains. right balance in reactant concentrations is vital to avoid
aggregation issues and achieve monodisperse nanopar-
Factors modulating biosynthesis ticles with the desired size [171].
The shaping and resizing of metallic and their oxide
nanomaterials seem to be influenced by their environ- Temperature effects on nanoparticle size and dispersity
mental conditions or altered by the presence of func- Temperature plays a crucial role in determining nanopar-
tional molecules [165]. Researchers have explored ticle size during synthesis. Higher temperatures tend to
modifying various synthesis parameters, such as ther- promote nucleation, leading to the formation of numer-
mal conditions, acidity/alkalinity, incubation duration, ous smaller nanoparticles. In contrast, lower tempera-
oxygenation, ionic strength, redox state, mixing pro- tures favor slower growth processes, resulting in fewer
portions, and irradiation, to optimize the production of but larger nanoparticles [172]. However, increasing the
nanoparticles. Both chemical and physical factors gov- overall reaction temperature accelerates the total reac-
ern the dimensions and morphology of nanoparticles. tion rate [173]. Notably, temperature exhibits contrasting
Achieving optimal conditions for metal ion concentra- effects on the size under sufficient versus insufficient pre-
tion, thermal environment, and acidity/alkalinity is cru- cursor conditions due to differing influences on nucle-
cial during the synthesis process [166]. The properties of ation and growth kinetic constants [174]. As temperature
nanoparticles formed via biological methods are signifi- increases, the enhanced reduction rate consumes most
cantly influenced by the incubation time of the reaction metal ions in nuclei formation, hindering secondary
medium. Prolonged incubation can lead to variations in reduction on preformed nuclei surfaces. Consequently,
characteristics, potentially due to particle aggregation elevated temperatures yield smaller, highly dispersed
or shrinkage [46]. Consequently, the particles’ self-life nanoparticles with improved yield [175].
or stability may impact their functional potential over
extended periods. Therefore, in addition to the synthe- Influence of pH on nanoparticle size, shape, and Colloidal
sis parameters like thermal conditions, acidity/alkalin- Stability
ity, substrate concentration, and exposure duration to The pH during nanoparticle synthesis exerts a profound
the substrate, the incubation time is a crucial factor that influence not only on the size but also on the shape of the
governs the characteristics and potential of biologically resulting particles [176]. The pH governs the local sur-
synthesized nanoparticles [167]. Adjusting these syn- face characteristics of nanoparticles [177] by facilitating
thesis parameters can modulate the rate of nanoparticle the protonation and deprotonation of molecular atoms
formation within cells and their eventual size to some during the nucleation and growth stages [178]. Notably,
extent. In the subsequent subsections, precursor concen- nanoparticles synthesized under lower pH conditions
tration, temperature, and pH will be further explained, exhibit less regularity in shape and a tendency toward
since these key parameters contribute to the optimiza- aggregation. However, by tailoring the pH conditions
tion and the efficiency of the biogenic process as they sig- during synthesis, nanoparticles with the desired size and
nificantly influence the nucleation rate, reaction kinetics, uniform shape can be produced [179]. Conversely, in the
aggregation, and stabilization of the desired nanoparticle alkaline pH range, nanoparticles form a well-dispersed
characteristics. cluster distribution in the colloidal stage, thereby pre-
venting aggregation [180].

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 12 of 40

Characterization of nanoparticles XRD pattern, the presence of these phases in the syn-
The physicochemical characterization of synthesized thesized TiO2-NPs can be determined. Vibrating sample
nanoparticles is a crucial step that demands meticulous magnetometry (VSM) evaluates their magnetic proper-
attention before their practical applications. Investigat- ties. Thermogravimetric analysis (TGA) assesses thermal
ing properties such as size, shape, surface area, homo- stability and quantifies components within nanoparticle
geneity, and stability provides valuable insights into samples. Zeta potential reflects the surface charge of
these nanoscale systems, enabling better control over NPs, influencing their suspension stability. Higher abso-
nanoparticle synthesis for commercial purposes [181]. lute values imply enhanced stability through electrostatic
Various characterization techniques are employed, each repulsion. Contact angle measurements determine the
serving a distinct function [182]. The color change test extent to which a liquid spreads on a solid surface, pro-
offers a simple visual indication of nanoparticle for- viding insights into the surface energy and hydrophobic-
mation. UV-visible spectrometry analyzes the optical ity of nanoparticles [186]. Moreover, the 2D shape and
properties and confirms the presence of nanoparticles. 3D shape were detected and investigated by atomic force
In the case of TiO2-NPs synthesized by bacteria, they microscopy (AFM), whereas the chemistry of TiO2-NPs
exhibit UV-Vis absorption peaks in the range of 300– surface, state of chemical and electronics within NPs, and
400 nm. Most of these peaks are observed between 350 elemental composition were assessed by X-ray photo-
and 400 nm, whereas in some cases, the peaks are found electron spectroscopy(XPS) [187].
below 350 nm. This variation in peak position is attrib- These techniques collectively offer comprehensive
uted to the presence of different biomolecules involved in characterization, enabling researchers to thoroughly
the synthesis process and the size and shape of the NPs, understand the synthesized nanoparticles and optimize
which affect their surface plasmon resonance. Fourier their synthesis for diverse applications.
transform infrared spectroscopy (FT-IR) elucidates the
chemical composition and identifies functional groups Biomedical and biotechnological applications of
that facilitate the stabilization and surface modification/ TiO2-NPs
capping of the synthesized TiO2-NPs. Within the rapidly evolving landscape of nanotechnol-
Electron microscopy techniques, including transmis- ogy, the realm of biomedical applications has witnessed
sion electron microscopy (TEM), high-resolution TEM remarkable progress fueled by extensive research endeav-
(HR-TEM), scanning electron microscopy (SEM), and ors. Nanoparticles have garnered significant attention
field emission-SEM (FE-SEM), provide detailed visualiza- due to their distinctive physicochemical properties,
tion and structural analysis of nanoparticles. Also, they which render them advantageous across a diverse array of
provide insights into the association of carbon-based bio- fields, including pharmaceutical formulations, diagnos-
molecules with the synthesized NPs [183]. In SEM, the tic tools, personal care products, and electronic devices
non-metallic regions appear darker due to electron defi- [188]. The unique characteristics inherent to nanomate-
ciency, while in TEM, these areas appear brighter com- rials, particularly their minuscule size ranging from 1 to
pared to the darker titanium element. Additionally, the 100 nm, heightened reactivity, and immense surface area,
elemental analyzer integrated with electron microscopy have unlocked novel avenues for exploration and innova-
facilitates the determination of the chemical composi- tion [189]. These properties facilitate their cellular entry
tion and purity assessment of the synthesized TiO2-NPs. and interaction with biomolecular and cellular pathways,
Complemented by data from (FTIR) and (XRD), electron rendering nanomaterials invaluable assets in the realm of
microscopy can reveal the presence and nature of bio- drug therapeutics [190](Fig. 3).
molecules associated with the TiO2-NPs [184]. TiO2 emerges as a highly versatile material, rendering
Energy-dispersive X-ray spectroscopy (EDX) mapping it an attractive choice across a wide array of applications
reveals elemental composition and distribution within due to its remarkable mechanical and photochemical
nanoparticles. Dynamic light scattering (DLS) mea- characteristics [191]. In the realm of cosmetics, TiO2
sures the size distribution and aggregation behavior of finds utility in formulating anti-aging and skin-brighten-
nanoparticles in solution [185]. ing products, harnessing its unique characteristics [192].
Powder X-ray diffraction (XRD) is crucial for charac- Similarly, its optoelectronic properties make it a valu-
terizing the synthesized TiO2-NPs as it determines their able component in the fabrication of devices and sen-
crystalline structure and phase composition. TiO2-NPs sors within the electronics industry [193]. Furthermore,
can exist in three different phases: anatase, rutile, and the medical field has embraced TiO2 for its potential in
brookite. The anatase phase exhibits a sharp peak near targeted drug delivery systems, antimicrobial activity,
two theta values (2θ) of 25–26°, while the rutile phase is treatment of cancer cells, biosensors, dental treatment,
identified by a peak around 2θ value of 27–28°. By ana- anti-inflammatory activity, etc. (Fig. 3). The following
lyzing the specific peak positions and intensities in the subsections comprehensively explore some biomedical

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 13 of 40

Fig. 3 Flowchart enumerates the biomedical and biotechnological applications of TiO2-NPs

and biotechnological applications of TiO2-NPs, high- (DPPH), within a reduced timeframe, demonstrating
lighting their remarkable versatility and vast potential their antioxidant capabilities (Table 6). TiO2-NPs are
across diverse domains. also reported to exhibit protective effects against reactive
oxygen species (ROS). Plant-mediated synthesis methods
Antioxidant activity result in TiO2-NPs with functional groups such as phe-
TiO2-NPs, especially those formed by plant extract that nols and tannins, contributing to their stabilization and
impaired various functional groups, showed high anti- enhanced antioxidant potential. These functional groups
oxidant activity to be utilized for decreasing the delete- present on the surface of the nanoparticles play a crucial
rious effects of free radical ions [194]. These TiO2-NPs role in scavenging free radicals and mitigating oxidative
were able to scavenge free radicals, such as 2,2′-casino- stress [195].
bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and In a study, the green synthesis of TiO2-NPs using
2,2-Diphenyl-1-(2,4,6-trinitrophenol) hydrazine-1-yl Psidium guajava (guava) leaf extract exhibited superior

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 14 of 40

Table 6 Antioxidant activity of green synthesized TiO2-NPs

TiO2-NPs Synthesized by Antioxidant Activity/ Assay used Reference
Psidium guajava (guava) leaf extract Superior to ascorbic acid/(TiO2-NPs:18.3 mg/g) [196]
Artemisia haussknechtii DPPH scavenging activity: (68.43% -500 µg/ml of TiO2-NPs) [197]
fruit peel agro-waste Dose-dependent antioxidant effects (DPPH, H2O2 free radical, NO, and reduc- [198]
ing power assays)
Lawsonia inermis 5-100 mg/ml of TiO2-NPs enhanced DPPH scavenging and 82% reduction in [199]
Leaf extract hydrogen-mediated hemolysis
Syringodium isoetifolium Strong antioxidant activity in DPPH and ABTS assays [200]
Tinospora Cordifolia 90% DPPH scavenging assay [201]
Trichoderma citrinoviridae 50–100 µg/ml of tested TiO2-NPs in DPPH were potent than standard gallic [138]
acid
Coleus aromaticus 100 µg/ml of TiO2-Nps tested caused 89% scavenging in DPPH assay [202]
Withania somnifera DPPH scavenging activity (68.43% at 500 µg/ml) [203]
Eclipta prostrata
Laurus nobilis (bay leaf ) DPPH 46.71% at 200 µg/ml and H2O2 of 58.45% at 50 µg/ml [203]
Terenna asiatica IC50 = 80.21 µg/µL in the DPPH assay [204]
Achillea wilhelmsii C. Koch ROS generation (215.4%) and decreased MMP (72%) [205]
Terminalia catappa bark extract 47% reduction in MDA content [85]
Pithecellobium dulce 52% and 45% for DPPH inhibition respectively [206]
Lagenaria siceraria
Leaves extracts
Malva parviflora extract 85% DPPH inhibition and 90% scavenging for ABTS. [207]
Tulbhagia violacea 50 µg/mL of TiO2-NPs showed IC50 = 32.7 in DPPH assay [82]
Leaf extracts
Limonia acidissima (ROS) release and (MMP) damage at 70 µg/ml of TiO2-NPs [208]
Peel extract
DPPH:2,2-Diphenyl-1-picrylhydrazyl; NO: Nitric Oxide; ABTS: 2,2’-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid); ROS: reactive oxygen species; MMP:
Mitochondrial Membrane Potential; MDA: Malondialdehyde

antioxidant activity compared to ascorbic acid. This Antimicrobial activity

enhanced antioxidant potential was attributed to the Antibacterial activity
presence of phenolic compounds in the aqueous leaf The indiscriminate use of antibiotics has resulted in
extract (85.4 mg/g) and the synthesized TiO2-NPs the emergence of multidrug-resistant bacterial strains,
(18.3 mg/g). which has become a significant cause for concern regard-
The phenolic content in the plant extract played a ing food safety and human health [209]. In the quest for
crucial role in the formation and stabilization of the novel antibacterial agents, metal oxide nanoparticles
nanoparticles, contributing to their remarkable antioxi- have garnered significant interest from researchers, with
dant properties [196]. Similarly, Artemisia haussknechtii biofilm formation being identified as a major factor con-
leaves synthesized TiO2-NPs with strong antioxidant tributing to antibiotic resistance [210]. Consequently,
properties. The synthesized NPs have been tested scientific investigations have shifted focus to explore the
through various assays, indicating a remarkable DPPH antimicrobial capabilities of nanoparticles composed of
scavenging activity of 68.43% at a concentration of metals and metal oxides.
500 µg/ml. The reducing power of TiO2-NPs was greater TiO2-NPs exhibit photocatalytic antimicrobial activity
than ascorbic acid (control), as determined by the reduc- when exposed to UV irradiation with a wavelength below
ing power assay to donate an electron [197]. In another 385 nm. However, the effectiveness of this antimicrobial
study, TiO2 nanoparticles, produced using fruit peel activity displayed by TiO2-NPs is contingent upon the
agro-waste, possess antioxidant potential. The study uti- thickness of the microbial cell surface [211]. TiO2-NPs
lized various scavenging assays such as DPPH free radi- exhibit antibacterial activity through the generation of
cal, H2O2 free radical, and NO, as well as the reducing reactive oxygen species (ROS), such as hydroxyl radicals
power assay. The results showed that TiO2 nanoparticles (OH-), superoxide anions (O2•-), and hydrogen peroxide
exhibited dose-dependent antioxidant effects compared (H2O2) (Fig. 4). These ROS induce oxidative stress on
to ascorbic acid, which was used as a control [198]. the bacterial cell membrane, inducing lipid peroxidation
in the plasma membrane’s unsaturated phospholipids.
Consequently, the bacterial membrane sustains dam-
age. Additionally, the photocatalytic activity of TiO2-NPs

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 15 of 40

Fig. 4 Various antibacterial mechanisms of biogenic TiO2-NPs

disrupts crucial biological processes within bacteria, biomedical and food packaging sectors (Table 7). Their
including respiration, oxidative phosphorylation reac- production encounters challenge due to the inherent
tions, and the maintenance of semi-permeability [212]. dissimilarities between the inorganic and hydrophilic
Figure 4 presents various antibacterial mechanisms of characteristics of the nanoparticles and the hydropho-
TiO2-NPs used to inhibit bacterial growth. bic nature of polymer matrices. Nonetheless, antimicro-
A research investigation examined the impact of bial nanocomposites derived from titania have garnered
TiO2-NPs on biofilm formation by Methicillin-Resistant substantial interest in recent times [215]. A research
Staphylococcus aureus (MRSA) using a tissue culture endeavor delved into the electrochemical fabrication
plate method (Table 7). Among the 30 isolates evalu- of silver-titania nanocomposites, striving to augment
ated, 22 demonstrated strong biofilm formation capabili- their photocatalytic characteristics while simultane-
ties, while 2 exhibited weak biofilm formation. The study ously amplifying their antifungal and antibacterial activi-
revealed that TiO2-NPs at a concentration of 500 µg/mL ties [216]. Another study focused on the development of
inhibited the growth of both strong and weak MRSA paraffin and silver-coated titania nanoparticles (TiO2/
biofilm formers, suggesting the potential of TiO2-NPs AgNPs) embedded in a polyethylene nanocomposite for
as viable antibacterial options [213]. In another study, food packaging purposes. Nanocomposite films were
TiO2-NPs were employed in combination with antibiotics fabricated by melt-blending 3% and 5% of TiO2/Ag NPs
ceftazidime and cefotaxime against multidrug-resistant into low-density polyethylene. The findings revealed
Pseudomonas aeruginosa. The bacterial samples were that the addition of 5% TiO2/Ag NPs resulted in a sub-
isolated from sputum, pus, and bronchoalveolar lavage. stantial decrease in bacterial growth [217]. Therefore, the
Upon 1-hour UV light exposure, a bactericidal effect was incorporation of TiO2 nanoparticles, whether used inde-
observed at TiO2-NPs concentrations exceeding 350 µg/ pendently or integrated with polymers, results in the sup-
ml. The minimum inhibitory concentrations of TiO2-NPs pression of microbial growth, thereby preventing food
were six-fold higher compared to the antibiotics. Con- spoilage and enhancing the shelf-life of food products
sequently, the combined application of antibiotics and [218].
TiO2-NPs synergistically augmented the antimicrobial
activity [214]. Antifungal activity
Recently, the synthesis of TiO2-NPs-based nano- Conventional fungicides and pesticides can be hazardous
composites is characterized by their high activity in the to health, making nanoparticles (NPs) a more desirable

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 16 of 40

Table 7 Summarizing the antimicrobial activity including antibacterial, antifungal, and antiviral of synthesized TiO2-NPs
TiO2-NPs nature Size Shape Target and finding References
Antibacterial
Terminalia chebula 56 nm tetragonal Decreased biofilm of St. mutans [219]
Rosa davurica 146 ± 3 nm irregular Bacterial biofilm inhibition of S. aureus and B. cereus [220]
Cynodon dactylon < 100 nm firmly Antibacterial effect against A. baumannii and St. aureus [221]
agglomerated
H. thelbiecea 40 nm spherical Bacterial cell membrane damage [222]
Ananos seneglensis 50 nm crystalline
Azadirachta indica Ficus benghalen- 10–33 nm tetragonal Bactericidal effect against Streptococcus mutans and Citro- [223]
sis Syzygium aromaticum crystalline bacter freundii
Ocimum americanum L. leaf 25 nm spherical Bactericidal action against Clostridium perfringens, S. paraty- [224]
phi and K. pneumoniae
Pleurotus djamor 31 nm spherical Antibacterial effect against Pseudomonas fluorescens and C. [144]
diphtheriae
Commercial < 50 nm NA Inhibited MRSA biofilm formation [213]
microemulsion 9 nm anatase structure Bactericidal effect on Pseudomonas aeruginosa [214]
Electrochemical TiO2 anatase spherical Enhanced antifungal and antibacterial activities [216]
(sacrificial anode) (15.6 nm)
Ag-TiO2
(15.6 nm)
sol-gel method 10–15 nm spherical Used in food packaging to prevent spoilage & decreased [217]
bacterial growth
Antifungal
Curcuma longa 92.6 nm anatase Increased resistance to damping off fungal disease by F. [225]
graminearum
Commercial 70–130 nm Anatase crystal C. albicans was inhibited by 65% [226]
Ball milling method 108–130 nm irregular hyphal lysis of Macrophomina phaseolina [227]
Bacopa monnieri < 100 nm homoge- Enhanced antifungal and antibiofilm activity against [228]
neous surface [Link] and [Link] compared to PVA alone
morphology
chemical 26 nm spherical Reduction of Candidal adhesion and biofilm formation [229]
Commercial 6 nm NA Biocidal against Aspergillus niger on Paulownia wood [230]
Commercial 50 nm thin homoge- Antifungal against wood-decaying fungi (Mucor circinelloides [231]
neous layer and Hypocrea lixii)
Pogostemon cablin 71.82 nm NA Antifungal effect at low MFCs [232]
Caricaceae (Papaya) shell extracts 15 nm Semispherical Antifungal activity against S. Sclerotiorums with improved [233]
seed germination
African oil palm 14.60 ± 0.44 nm agglomerated Fusarium solani growth inhibition [234]
hemispherical
Trichoderma harzianum 431 ± 87 spherical High chitinase activity against Sclerotinia sclerotiorum [139]
Trianthema portulacastrum 30–60 nm Granule-like Fungicidal effect against Ustilago tritici [83]
Chenopodium quinoa shapes
ultrasonic NA Spherical with Inhibited spores’ germination of F. graminearum [235]
aggregation
Caesalpinia pulcherrima 20–27 nm spherical Superior anticandidal activity at low MICs [236]
flower extract
Antiviral
Sonochemical method 8 nm tetragonal Antiviral treatment of Newcastle disease virus (NDV) [237]
Chemical hydrolysis 4–10 NA Viral treatment against H3N2 Influenza virus [238]
Solid state reaction method 1 m²/g irregular Antiviral filtration against H1N1 (face masks) [239]
chemical impregnation method NA amorphous Antiviral properties against H1N1 and SARS-CoV-2 [240]
(COVID-19)
sol-gel method 50–100 nm tubular Potent anti-SARS-CoV-2 activity [241]
electrochemical anodization 10 nm tubular Electrochemical sensor for rapid detection of SARS-CoV-2 [242]
technique
Commercial 50 nm spherical Antiviral activity against Human Papillomavirus HPV [243]
Chemical adsorption method 1–100 nm Anatase Anti SARS-coV-2 [244]

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 17 of 40

Table 7 (continued)
TiO2-NPs nature Size Shape Target and finding References
Sol gel method 20 nm hemispherical Antiviral Activity Against Tobacco mosaic virus (TMV) in [245]
Pepper plants
Hydrolysis chemical method 5–6 nm anatase Increased silkworm to Bombyx mori nucleopolyhedrovirus [246]
(BmNPV)

option as they exhibit fewer adverse effects and a more ATP levels, depriving the cells of their primary energy
favorable therapeutic index, indicating a better safety currency and impairing cellular functions [253]. Lastly,
margin [247]. Recently, TiO2-NPs have revealed fungi- they can interact with and damage DNA, leading to
cidal effects against different human and plant patho- genetic alterations and impaired replication [254]. They
genic fungi such as Fusarium oxysporum [248], Fusarium also disrupt protein structure and function and interfere
graminearum [225], the opportunistic human pathogen with mitochondrial processes, further contributing to
Candida albicans [226], Macrophomina phaseolina [227] cellular dysfunction and growth inhibition (Fig. 5).
(Table 7). Also, the researchers synthesized nanocompos- Laboratory and field studies on heritage buildings have
ites comprising polyvinyl alcohol (PVA) and TiO2-NPs. also demonstrated the efficacy of TiO2 as an effective bio-
These nanocomposites were evaluated for their anti- cidal agent against lichens and phototrophic microbes,
fungal properties against two fungal strains: Candida as well as a preventive material against the biodeteriora-
albicans (ATCC 14053) and Penicillium chrysogenum tion of buildings [255]. Furthermore, the application of
(MTCC 5108); the results demonstrated that the PVA- TiO2-NPs has been proposed as a protective measure for
TiO2 nanoparticle biofilms showed enhanced antifungal culturally significant buildings to mitigate the biodeterio-
activity potential comparing to PVA alone. The incorpo- ration of mortars caused by microbial growth [256]. For
ration of TiO2 nanoparticles into the PVA matrix con- example, in one study, researchers evaluated the effects
ferred superior antifungal efficacy to the nanocomposite of TiO2-NPs against the fungal mold Aspergillus niger
material [228]. on the surface of Paulownia wood. The results showed
Candidiasis, caused by Candida species, is an opportu- that the treatment with TiO2-NPs effectively suppressed
nistic fungal infection affecting various mouth, skin, and the growth of the fungal mold [230]. In another study,
genitourinary tracts [249] and can cause life-threatening the antifungal effects of TiO2-NPs were assessed against
bloodstream candidemia. It has a high global annual wood-decaying fungal species, including brown rot
incidence of approximately 4 × 106 cases with a mortal- (Mucor circinelloides) and white rot (Hypocrea lixii) fungi
ity rate of about 40%, resulting in a substantial economic [231]. This suggests that TiO2-NPs possess remarkable
burden due to treatment costs and prolonged hospital- antifungal properties and can be employed as an effective
ization [250]. It has been reported that metal and their protective measure against fungal growth and deteriora-
oxide nanoparticles such as (TiO2-NPs, Ag-NPs, Cu-NPs, tion in various wood-based applications.
ZnO-NPs) exhibited antifungal activity against Candida However, it is noteworthy that the antifungal activity
species through various mechanisms that synergistically exhibited by TiO2-NPs appears to be relatively weaker
contribute to the inhibition and eradication of these fun- compared to their antibacterial action. This discrepancy
gal pathogens. Firstly, those nanoparticles release ions can be attributed to the fundamental differences in the
that disrupt crucial cellular processes within the fungal structural composition of the cellular envelopes between
cells [229]. Additionally, they induce oxidative and nitro- fungi and bacteria. Fungal cells possess a more robust
sative stress by generating reactive oxygen and nitrogen and complex cell wall structure, which may render them
species, causing deleterious effects on cellular biomol- less susceptible to the antifungal mechanisms of TiO2-
ecules such as lipids, and nucleic acids [39]. Furthermore, NPs compared to bacterial cells with their relatively
these nanoparticles can directly interact with and com- simpler cell envelope architecture [257]. Table 7 summa-
promise the structural integrity of the fungal cell mem- rizing examples for antifungal activity of biogenic TiO2-
brane and cell wall, compromising their barrier functions NPs against various pathogenic fungi.
and potentially causing leakage of cellular contents [251].
Remarkably, certain nanoparticles can inhibit the activity Antiviral activity
of essential enzymes involved in various metabolic pro- Nanoparticles like Ag, TiO2, and carbon nanotubes pos-
cesses, hampering the growth and survival of fungal cells sess antiviral properties. These properties are employed
(Fig. 5). Moreover, they can modulate the expression of to showcase the antiviral mechanisms, which include
specific genes involved in stress response, cell wall bio- inhibiting the virus from binding to cells and facilitating
synthesis, and virulence factors, effectively regulating the the breakdown of the viral capsid [258]. Several factors
fungal cell’s ability to thrive [252]. Also, they can deplete influence the antiviral capabilities of NPs, including their

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 18 of 40

Fig. 5 Suggested antifungal mechanisms produced by TiO2-NPs

behavior in air and water environments when studied in and the virus [238]. Additionally, TiO2-NPs have shown
vitro. antiviral efficacy against the H9N2, the avian influenza
Notably, TiO2-NPs have exhibited superior antivi- virus. Notably, the replication of two other influenza
ral activities against the Newcastle disease virus (NDV) strains, H5N1 and H1N1, was effectively inhibited by the
when assessed at doses ranging from 6.25 to 100 µg/ml application of DNA-tagged titanium nitride nanopar-
(Table 7). The mechanism behind this antiviral activity ticles (TiNPs) [211]. The H1N1 influenza virus, with
involves the potential of TiO2-NPs to destroy the lipids its high frequency of genetic polymorphism [260, 261]
present in the viral envelope through the generation of that caused the 2009 pandemic, also posed an increased
ROS, a process known as G-sol. The glycoprotein spikes risk of subsequently developing type 1 diabetes in chil-
on the viral surface, which are known to be harmful in dren infected by it [249]. An interesting study has docu-
facilitating infections, are restricted by an adjunct mech- mented the potent antiviral capability of a TiO2-modified
anism, preventing the virus from binding and infecting hydroxyapatite composite (HA/TiO2) against the H1N1
cells. Consequently, TiO2-NPs represent a promising Influenza A Virus when exposed to UV light irradiation.
platform for the treatment of Newcastle disease virus Notably, these composites exhibit potential for antimi-
infections [237]. Also, TiO2 nanostructures were uti- crobial filtration applications, rendering them suitable
lized to treat faba bean crops infected with the broad for use in face masks to combat such highly mutable
bean stain virus (BBSV). The faba bean plants treated influenza strains [239]. Also, a novel nanocomposite
with these NPs demonstrated a significant reduction in comprising TiO2-NPs and polylysine (PL)-containing
the severity of the viral disease compared to untreated oligonucleotides, termed TiO2⋅PL–DNA. This nanocom-
plants. This superior antiviral effect was observed within posite exhibits antiviral properties against various sub-
two weeks after the plants were infected with the broad types of the influenza A virus, including H1N1, H5N1,
bean stain virus [259]. and H3N2 [240].
Further studies have highlighted the antiviral poten- Investigations have demonstrated the remarkable
tial of TiO2 nanoconjugates and nanoparticles against disinfection capability of titanium dioxide nanotubes
various influenza viruses. In one investigation, TiO2- (TiO2-NTs) against SARS-CoV-2. These nanostructures
NPs displayed antiviral capabilities against the H3N2 exhibited potent anti-SARS-CoV-2 activity at extremely
influenza virus strain, with the proposed mechanism low concentrations in vitro, coupled with negligible
involving direct interaction between the nanoconjugates cytotoxicity and an insignificant selectivity index (CC50/

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 19 of 40

IC50 ≤ 1). Moreover, they displayed excellent antiviral Under UV light exposure, the TiO2 surface facilitates
efficacy at very low concentrations (IC50 = 568.6 ng/mL). the decomposition of ambient oxygen and water into
Consequently, it was concluded that these TiO2 nano- ROS, which act as highly oxidizing or reducing agents,
structures are well-suited for use as coatings, serving as leading to the decomposition of organic and microbial
potent disinfectants to combat SARS-CoV-2 [241]. While matter. Furthermore, modified TiO2 has demonstrated
in another study, researchers developed an electrochemi- remarkable utility in visible light activity, enabling indoor
cal sensor based on TiO2- NTs for the rapid detection or outdoor disinfection applications. This photocatalytic
of SARS-CoV-2. The surface engineering of these TiO2 disinfection effect has also shown promising potential as
nanostructures has been proposed as a strategy to tailor an antiviral photocatalyst for controlling various viruses
their potential functionality, enabling real-world applica- [211]. ROS, such as OH•, O2•-, and H2O2, are produced at
tions in combating the SARS-CoV-2 virus [242]. Inter- the surface of TiO2 due to UV activation (Fig. 6).
estingly, a study explored the potential of TiO2 coatings These highly reactive species possess the capability to
in inactivating SARS-CoV-2 through time-dependent, degrade the capsid proteins of non-enveloped viruses as
TiO2-mediated photocatalytic reactions. Transmission well as the envelope proteins and phospholipids of envel-
electron microscopy (TEM) revealed microstructural oped viruses. The degradation of these crucial viral com-
changes in the SARS-CoV-2 virus upon interaction with ponents leads to the leakage of genetic material, resulting
the coating. The antiviral activity, assessed in aerosol in the subsequent degradation of nucleic acids. Conse-
and liquid forms, exhibited up to 99.9% effectiveness quently, this process culminates in the eventual inactiva-
after 20 min of exposure. The mechanistic effects on the tion of the viral particles, rendering them non-infectious
SARS-CoV-2 virion included decreased virion count, and incapable of replication or causing further infections
increased virion size, and reduced particle surface spike [263].
structure. Further analyses using western blotting and
RT-qPCR investigated the photocatalytic damaging of Anticancer activity
viral proteins and genomes, respectively. The study con- Nanotechnology has gained prominence in cancer treat-
cluded that TiO2-induced photocatalytic reactions hold ment and diagnosis due to the severe side effects associ-
promise for disinfecting SARS-CoV-2 and other emerg- ated with traditional chemotherapeutic agents, which
ing infectious agents in human habitats [262]. exhibit cytotoxicity on healthy cells. The application

Fig. 6 Antiviral mechanisms of TiO2-NPs based on secretion of ROS

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 20 of 40

of nanotechnology offers a more targeted and precise (DEN)-induced hepatocellular cancer animal model. The
approach, minimizing harm to normal cells while effec- study’s findings revealed that the surface-modified Pacli-
tively combating cancer [264]. Nanotechnology research taxel (PAC) attached to TiO2-NPs exhibited superior
in cancer therapy focuses on developing nanostructures anticancer activity compared to PAC alone. Significantly,
capable of delivering and releasing drugs in a targeted the surface modification of TiO2 with FA in the TiO2-
manner. Pursuing targeted drug delivery and controlled PEG-FA-PAC nanocomposite facilitated targeted deliv-
drug release represents the primary approach to aug- ery to liver cancer cells overexpressing FA receptors. This
menting therapeutic efficacy while mitigating adverse targeted approach resulted in increased accumulation of
effects. PAC-NPs at the cancer site, thereby reducing the drug’s
TiO2 nanostructures, renowned for their high bio- toxicity. Moreover, the TiO2-PEG-FA-PAC nanocompos-
compatibility, tunable drug release capabilities, and ite demonstrated a reduction in cell viability correlated
minimal toxicity, are widely acknowledged as promising with concentration. when tested on HepG2 liver cancer
candidates for enabling targeted delivery and controlled cells, further underscoring its potent anticancer effects
release of conventional chemotherapeutic agents, con- [270].
sequently enhancing their clinical therapeutic impact Another study reported the enhanced anticancer activ-
[265]. Furthermore, TiO2-NPs have demonstrated their ity of FA-TiO2-NPs against MG63 osteosarcoma cells.
efficacy as drug carriers, facilitating targeted delivery Compared to unconjugated nanoparticles, the FA-TiO2-
of chemotherapeutics like doxorubicin, cisplatin, and NPs exhibited a two-fold lower IC50 value, indicating
temozolomide [266]. The subsequent sections provide a improved cytotoxicity and efficacy in inhibiting can-
comprehensive review of significant research endeavors cer cell growth. The study also explored the apoptosis-
that have explored the utilization of TiO2 nanostructures inducing effects of FA-TiO2-NPs on osteosarcoma cells.
in targeted drug delivery mechanisms and diverse con- Treated cells displayed hallmark apoptotic features: con-
trolled release systems, shedding light on their potential densed chromatin, surface membrane vesiculation, and
in cancer treatment. cell volume reduction [271].
Remarkably, Annexin V/PI apoptosis assay unveiled a
Target drug delivery significantly higher percentage (38%) of cells undergoing
The primary objective of drug delivery systems is to early and late apoptosis upon FA-TiO2-NP exposure com-
enhance therapeutic efficacy while minimizing adverse pared to only 16% with unmodified TiO2-NPs. Moreover,
effects through appropriate drug encapsulation. These cell cycle analysis revealed an augmented sub-G1 cell
systems have been designed with varying structures, cycle in FA-TiO2-NP-treated cells, signifying escalated
such as spherical, capsular, and porous configurations, to reactive oxygen species (ROS) production and height-
achieve specific drug release profiles [267]. In this con- ened apoptosis induction. A parallel study demonstrated
text, the drug can be situated on the surface of TiO2-NPs, that the delivery of doxorubicin via TiO2 nanocompos-
potentially leading to a controlled and sustained release ites facilitated enhanced intracellular drug retention and
pattern. TiO2-NPs are functionalized with specific mol- cellular internalization in multidrug-resistant MCF-7/
ecules that possess targeting capabilities toward desired ADR breast cancer cells, effectively circumventing the
cells, allowing them to localize within tissue-specific P-glycoprotein-mediated efflux mechanism responsible
cells and thereby enhancing their efficiency. These tita- for drug resistance [272]. Researchers in another study
nium nanoparticles have been employed as nanocarriers developed a pH-responsive drug delivery system by mod-
for the delivery of various drugs, including sodium phe- ifying TiO2-NPs with hyaluronic acid (HA) and loading
nytoin, valproic acid, temozolomide, and daunorubicin them with the chemotherapeutic agent cisplatin for ovar-
[268]. Initially, the drug release exhibited a burst pattern, ian cancer treatment. This nanoformulation facilitated
followed by a controlled release over an extended period. enhanced cisplatin accumulation within A2780 ovarian
Consequently, TiO2-NPs demonstrated the ability to cancer cells via endocytosis, exhibiting significant anti-
deliver drugs with enhanced efficacy, optimized dosing, cancer effects [273].
and controlled and sustained release profiles while simul-
taneously reducing toxicity levels. Controlled drug release in cancer therapy
The surface modification of nanocarriers plays a crucial In an effort to enhance therapeutic efficacy and mitigate
role in enhancing the specificity of drug delivery [269]. the undesirable side effects of the chemotherapeutic drug
A research study explored this concept by modifying doxorubicin. Doxorubicin was encapsulated within TiO2-
TiO2-NPs with polyethylene glycol (PEG), followed by NPs, forming DOX-TiO2-NPs. Subsequent evaluation
the attachment of folic acid (FA) and the standard anti- of the anticancer potential of DOX-TiO2-NPs revealed
cancer drug Paclitaxel (PAC). These anticancer proper- an increased cytotoxic activity against the SMMC-7721
ties were investigated in vivo using a diethylnitrosamine hepatocarcinoma cell line, as demonstrated by the MTT

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 21 of 40

assay [274]. The ratio of Bax/Bcl-2 protein was increased exposing cancer cells to high ROS levels could serve as
upon DOX-TiO2-NP treatment which indicates the a novel target for killing cancer cells without deleterious
endocytosis uptake of doxorubicin, leading to caspase- effects on normal ones [281].
apoptotic processes [275]. It is worth highlighting that
TiO2-NPs displayed minimal cytotoxicity when used Photodynamic therapy of cancer
independently at a concentration of 10 µg·mL− 1, with cell Cancer cells are subjected to laser light to produce a pho-
viability of 95%, indicating their potential for safe bio- tosensitive agent at a specific wavelength called photo-
medical applications [274]. dynamic therapy (PDT). Unlike other treatments such as
surgery, radiation, and chemotherapy, PDT is considered
Stimuli-controlled drug release a secondary and highly promising non-invasive modal-
Stimuli-responsive systems are designed to release drugs ity for cancer therapy [282]. It offers an encouraging
in response to specific signals. These signals can be clas- approach as a supplementary treatment option for can-
sified into two categories: internal and external stimuli. cer patients [283]. The inorganic nature of TiO2 endows
Among the benefits of this system are reducing the side it with the capability to produce ROS when exposed to
effects of used treatment drugs and increasing the drug’s UV light in aqueous environments (Fig. 7). This ROS
biocompatibility. They facilitate controlled and tissue- generation, which subsequently triggers cell death, posi-
specific drug release, contributing to improved thera- tions TiO2 as a promising candidate for PDT, a therapeu-
peutic outcomes [276]. light emerges as an intriguing tic approach utilized in the treatment of diverse diseases
external trigger for enabling the controlled and timed [284]. The versatility of TiO2-NPs, their nanocompos-
release of chemotherapeutic agents from delivery sys- ites, and hybrid biomolecular forms has been extensively
tems, owing to its capacity for precise spatial and tempo- explored, revealing their potential as photosensitizing
ral targeting [277]. In this context, TiO2 nanostructures agents for cancer treatment and combating antibiotic-
have garnered substantial interest as photoactive drug resistant bacterial infections [285]. When TiO2-NPs are
carriers. In addition to their photoactive nature, TiO2- exposed to UV light with a wavelength below 385 nm,
NPs are preferred as drug carriers due to their character- photoexcited electrons and holes are generated. Subse-
ized high surface area, durability, and accessibility. quently, these photoexcited electrons and holes can react
In a study, a mesoporous TiO2 shell as a core-shell with OH− or H2O, forming oxidative radicals capable of
structure for near-infrared light-triggered drug deliv- destroying microorganisms and tumor cells (Fig. 7).
ery. Doxorubicin was loaded into the porous TiO2 shell, The production of ROS by TiO2-NPs has been reported
and hyaluronic acid (HA) capping was performed. The to function as effector signaling mediators in the
authors reported that the cell viability was decreased at p53-dependent apoptotic pathway. Upregulated expres-
drug release at low concentrations, indicating its prom- sion of cytochrome c, cleaved caspase-3, and PARP, as
ise for cancer therapy [278]. Also, colloidal TiO2-NPs observed through western blot analysis, further indi-
were utilized as carriers for light-controlled delivery of cated the induction of apoptosis via caspase activation
a ruthenium complex drug to melanoma cancer cells. (Fig. 7), highlighting the therapeutic promise of surface-
This system exhibited a faster drug release profile upon engineered TiO2-NPs. The generated ROS can damage
UV light exposure compared to visible light illumina- the mitochondrial membrane and its functionality, sub-
tion. Furthermore, cell death increased when exposed to sequently initiating the mitochondrial release of cyto-
UV light as opposed to red light. The authors proposed chrome c into the cellular matrix, thereby triggering the
that both the TiO2-NPs and the ruthenium complex intrinsic apoptotic cascade [286].
could act as photosensitizers, generating reactive oxygen
species and inducing cell death [279]. In another study, Theragnostic
DOX was loaded onto TiO2-NPs, which were coated with The convergence of diagnostic and therapeutic
polymeric phenylboronic acid (PBA) through a boronic approaches has given rise to a new field known as therag-
ester bond. These nanoparticles exhibited high tumor- nostic. These strategies enable researchers and clinicians
targeting ability due to the specific interaction between to precisely modulate molecular signaling cascades at the
PBA and sialylated epitopes on tumor cells. Additionally, molecular level, which is particularly relevant in the con-
ultrasound irradiation could generate ROS, leading to the text of PDT [287]. In this regard, nanocomposites com-
release of DOX from the nanoparticles via cleavage of the prising zinc(II) phthalocyanine (MCZnPc) and MCZnPc
boronic ester bond [280]. anchored onto TiO2-NPs labeled with radioactive iodine
Studies show that increased oxidative stress from ele- (125I) were synthesized to evaluate their potential for
vated tumor ROS levels can be more damaging to cancer anticancer applications. The researchers evaluated the
cells. ROS generation in the tumor environment trig- efficacy of MCZnPc and MCZnPc-TiO2 nanocompos-
gers desirable apoptotic cell death. Therefore, selectively ites against HeLa (human cervical cancer) and EMT6

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 22 of 40

Fig. 7 Mitochondrial dysfunction and apoptotic pathways triggered by TiO2-NPs

(mouse mammary cancer) cell lines. After a 3-hour incu- alterations within cellular environments, as well as the
bation period in the dark, the cells were exposed to light measurement of hazardous environmental contaminants
at a wavelength of 684 nm. The labeled nanocomposites [292] and optimizing electrochemical biosensors for the
exhibited enhanced cellular uptake, and the study dem- precise detection and quantification of toxic chemicals in
onstrated the theragnostic potential of TiO2-NPs for can- food products to boost food safety protocols [293]. Bio-
cer treatment [266]. In a recent study, the incorporation sensors derived from nanomaterials possess remarkable
of samarium as a dopant into TiO2-NPs was shown to capabilities, enabling highly sensitive and rapid detection
enhance the radiosensitivity and cellular toxicity of can- of biological entities [294]. The recent fascination with
cer cells. The doped nanoparticles exhibited increased innovative hybrid systems combining biomolecules and
X-ray absorption, leading to the generation of higher TiO2 nanostructures has resulted in substantial achieve-
ROS upon irradiation compared to undoped TiO2-NPs ments in manufacturing bio-nano hybrid devices, such
[288]. TiO2-based platforms have been utilized for the as biomolecule-sensitized solar cells (BSSCs) and photo-
detection of small biomolecules, cancer cells, and patho- electrochemical cells (PECs) [295].
gens in blood samples through various techniques such In addition, the biosensors field was advanced through
as label-free microfluidic immunosensors, photoelectro- the integration of TiO2-NPs and biomolecules to form
chemical biosensors, field-effect transistors, and ampero- thin films that monitor patients’ responses to medica-
metric methods [289]. tion or surgical treatment [296]. However, for a biosensor
Additionally, Nanoconjugates composed of TiO2, poly- to be commercially viable, it must meet specific criteria:
ethyleneimine (PEI), and folic acid (FA) have been devel- cost-effectiveness, user-friendliness, sensitivity and accu-
oped to construct a controlled drug delivery system racy, rapid response times, and the ability to be manufac-
regulated by near-infrared (NIR) laser irradiation [290]. tured efficiently with high selectivity rates [297].
The induced and sustained delivery of TiO2 nanoparti- TiO2 nanostructures have been utilized to fabricate a
cles facilitated by X-ray exposure can generate electron- diverse range of sensing devices, encompassing humid-
vacancy pairs, leading to the structural degradation of ity, oxygen, and hydrogen sensors. These nanoscale semi-
organic linkers within the nanoconjugates [291]. conductors have proven their efficacy as exceptional
electrode materials within biosensors, owing to their
Biosensors distinctive properties. Such properties include a porous
Nanosensors are minute yet highly perceptive devices, architecture, providing a vast specific surface area, cou-
with at least one of their sensing dimensions measuring pled with outstanding biocompatibility [298]. TiO2 pos-
around 100 nm or less. These sophisticated instruments sesses the capability to function as an immobilizing
play a crucial role in detecting and evaluating intricate matrix, engaging in reactions with the amine and car-
physical and chemical transformations. Moreover, they boxyl groups of enzymes while concurrently preserving
enable the observation of biochemical and biomolecular their biocatalytic activity [299].

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 23 of 40

Recently, nanocomposites, including TiO2-NPs, are and holes upon exposure to light. Secondly, a catalytic
used in biosensors. For instance, nanocomposites con- counter electrode (CE) coated with an electron trans-
taining graphene oxide/TiO2-nanowires/chitosan were fer material is present. Finally, an electrolyte is placed
used to monitor the sequences of specific genes in Vib- between the two electrodes, facilitating the shuttling of
rio parahaemolyticus. The incorporation of TiO2 nanow- holes to the counter electrode [296]. In PEC biosensors,
ires in the nanocomposite significantly increased the the detection principle relies on photon-to-electricity
interfacial area, thereby enhancing its sensing capabili- conversion. The process involves photon absorption by
ties [300]. TiO2-based biosensing of target analytes typi- the semiconductor, generating electron-hole pairs that
cally employs either electrochemical techniques (such as separate and transfer to the working electrode and elec-
amperometric and potentiometric processes) or photo- trolyte, respectively. The presence of the target analyte
electrochemical (PEC) methods. This alteration manifests modulates the photocurrent generated, which correlates
as a detectable current signal in the case of amperometric directly with the analyte’s concentration in the sample,
detection or a detectable potential or charge accumula- enabling quantitative biosensing [302]. In TiO2-based
tion for potentiometric detection [301]. PEC biosensors, photoelectrochemical biosensors, the transducer mate-
which leverage the photoelectric effect, have emerged as rial can take one of three forms: (a) consisting solely of
a promising class of electrochemistry-based biosensing TiO2, (b) a hybrid combination of TiO2 and inorganic
devices. These innovative sensors combine the benefits semiconductors, or (c) a composite material where TiO2
of both optical and electrochemical detection techniques, is combined with other substances.
making them highly attractive for various applications Diverse biomolecules like enzymes and DNA have
[302]. served as bio-recognition elements. When DNA is cou-
Typically, a titanium dioxide-based PEC biosensing pled with TiO2, the presence of target DNA results in the
platform consists of three main components (Fig. 8). formation or cleavage of double-stranded DNA, leading
Firstly, a nanostructured TiO2 layer is fabricated on a to a direct or indirect change in the light-harvesting per-
conductive surface, acting as the working electrode (WE). formance of the working electrode. Consequently, this
This facilitates the generation and movement of electrons alteration impacts charge separation and the generated

Fig. 8 TiO2-based photoelectrochemical (PEC) biosensing platform

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 24 of 40

photocurrent [303]. Quantum dots with narrow band well as more advanced scanning techniques such as laser
gaps can sensitize TiO2-NPs and achieve energy band ablation, inductively coupled plasma mass spectrometry
modulation. Cadmium sulfide nanoparticles (CdS-NPs), (ICP-MS), and matrix-assisted laser desorption/ioniza-
possessing a narrow band gap of 2.4 eV and a broad exci- tion mass spectrometry (MALDI-MS) [308]. Enhance-
tation spectrum, are well-suited as sensitizing materials. ments in diagnostic techniques lead to preliminary
The excitation of CdS -NPs leads to charge separation, treatment and improved recovery prospects for patients.
facilitating the transfer of conduction band (CB) elec- Among nanoparticles, titanium dioxide nanoparticles
trons to the conduction band of TiO2. A recent hybrid TiO2-NPs are extensively researched and employed in
material combining CdS QDs and TiO2-NTs has found diagnostic methods like MRI and CT scans, serving as
application in the sensitive detection of prostate-specific contrast agents. Notably, TiO2-NPs become activated
antigen (PSA), a crucial biomarker for prostate can- upon irradiation, enabling them to act concurrently as
cer [304]. In this application, the CdS QDs were loaded diagnostic tools and therapeutic agents.
onto the TiO2-NTs, which acted as transducers. The A research study examined the image contrast capabili-
sensitization of TiO2 with CdS broadens the excitation ties of TiO2-NPs using magnetic MRI and CT scanners.
wavelength range and enhances the photoelectric perfor- A clear distinction in imaging was detected between
mance of the TiO2 electrodes. Additionally, the coupling the control samples and those containing TiO2-NPs on
between CdS and TiO2 reduces the recombination of T2-weighted MRI images. This finding suggests that
photo-generated electrons and holes, collectively leading TiO2-NPs could potentially serve as a novel theragnostic
to higher conversion efficiency [305]. When PSA is pres- agent, offering both radio-sensitizing properties for ther-
ent, an immune-sandwich assembly facilitates the attach- apeutic applications and radiological diagnostic function-
ment of an immune-gold-labeled alkaline phosphatase ality due to chemical modifications on their surface [309].
(ALP) to the CdS QDs/TiO2 NTs electrode. ALP can cat- According to the findings, in-depth investigations were
alyze the hydrolysis of ascorbic acid 2-phosphate (AAP) carried out on in-situ tagging methodologies for fluores-
in situ, thereby generating ascorbic acid (AA) for efficient cence microscopy to mark the TiO2-NPs internalized by
electron donation. The results indicate that an increase cells. The initial technique involved utilizing fluorescent
in PSA concentration leads to an improved photocurrent biotin and fluorescent streptavidin to tag the nanopar-
response. ticles before and after cellular uptake. Conversely, in the
PEC biosensors can be combined with alternative second approach, copper-catalyzed azide-alkyne cyclo-
detection techniques, enabling the development of bio- addition was employed for labeling and identification
sensors tailored to specific properties and behaviors. In of azide-conjugated TiO2-NPs. Moreover, synchrotron
a study, dye-sensitized solar cells (DSSCs) utilizing TiO2 X-ray fluorescence microscopy (XFM) was utilized to
nanoparticulate films as the photoanode were integrated detect TiO2-NPs. The results showcasing TiO2-NPs by
with a colorimetric DNA detection approach to sense a XFM exhibited remarkable alignment with the location
particular DNA sequence from Mycobacterium tuber- of optical fluorescence as detected by confocal micros-
culosis, resulting in a disposable biosensing device [306]. copy [310].
In another study, a nanocomposite consisting of g-C3N4 TiO2-NPs can be readily synthesized and modified,
and TiO2 nanosheets was employed to fabricate a photo- such as by incorporating europium(III) ions. Addition-
electrochemical biosensor capable of detecting glucose ally, hollow TiO2 nanoshells serve as viable two-photon
with an impressive detection limit of merely 0.01 mM. nanoprobes. When coated with polyethyleneimine, these
The two-dimensional TiO2 nanosheets, possessing a high nanoparticles demonstrate an affinity for binding to
specific surface area, demonstrated promising potential HeLa cervical cancer cells, enabling their detection [311].
for accommodating a substantial glucose oxidase load- In another research, a nanostructure comprising a silver
ing. Furthermore, to enable visible light excitation of core surrounded by a silica shell and an outer mesopo-
the photoelectrochemical biosensor and avoid potential rous titania coating (Ag@SiO2@mTiO2) was developed.
deactivation by UV radiation, g-C3N4 was incorporated The metallic silver core served to enhance fluorescence
to minimize the nanocomposite’s bandgap [307]. signals [312].

Biomedical imaging Agriculture applications

There is a wide range of imaging techniques that can be Contemporary advanced technologies, including aquatic
utilized for scientific research and biomedical applica- farming, solar-powered greenhouses, genetic engi-
tions. These include spectroscopy methods like infrared neering, multi-layer crop production, and anti-mold
(IR) spectroscopy and Raman spectroscopy, nuclear mag- chemicals, have conferred substantial benefits to the agri-
netic resonance imaging (MRI), radio-imaging using spe- cultural sector by enabling maximum yield production
cific nuclides, computed tomography (CT) scanning, as and the cultivation of off-season crops. However, these

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 25 of 40

technologies have also contributed to significant health TiO₂-NPs against S. littoralis larvae at all the concentra-
and environmental concerns due to the improper utiliza- tions tested [316]. In another study, TiO₂-NPs were tested
tion of fertilizers and pesticides [313]. Nanotechnology individually and in combination with ZnO-NPs to evalu-
is a rapidly evolving interdisciplinary domain focused on ate their insecticidal efficacy against Bactericera cocker-
developing innovative technological tools to maximize elli nymphs through laboratory and greenhouse studies
crop yields and enhance plant protection. on tomato plants. While leaf immersion bioassays were
The underlying strategy involves boosting plants’ ability conducted in the laboratory, direct plant spraying was
to absorb essential nutrients more efficiently. Among var- employed in the greenhouse. Results showed that TiO₂-
ious nanomaterials, extensive research efforts have been NPs alone (100 ppm) and combined with ZnO -NPs
concentrated on exploring the agricultural applications of (250 ppm) caused 99% and 100% mortality, respectively,
TiO2-based nanomaterials due to their distinctive struc- after 96 h of treatment in the laboratory. However, in the
tural properties, chemical stability, hydrophilic nature, greenhouse experiment, TiO₂- NPs (500 ppm) and TiO₂ +
and environmental compatibility [314]. The subsequent ZnO NPs (250 ppm) resulted in only 32% and 23% mor-
section highlights the applications of nanopesticides and tality, respectively [317]. Also, a different study explored
nanofertilizers derived from TiO2-NPs. the insecticidal potential of TiO₂-NPs prepared via a
green synthesis method using the aqueous leaf extract
TiO2-NPs as nanopesticides of Pouteria campechiana. The findings revealed that at a
Maintaining crop health by preventing diseases and pest concentration of 900 µg/ml, the larvae and pupa of Aedes
infestations is a persistent challenge that drives the cre- aegypti were dead [318].
ation of novel solutions and agents. Chemical pesticides TiO2-NPs loaded with fluorine and nitrogen were used
are employed to manage or eliminate microorganisms, to inhibit the Fusarium oxysporum growth in tomatoes
unwanted plants, insects, and fungi. Nevertheless, exces- under visible light conditions by destroying of fungal cell
sive pesticide usage can lead to severe health implica- wall. The synergistic effects of TiO₂-NPs and the attached
tions. Nanoparticles or nanoformulations of pesticides N & F resulted in a stronger toxic impact on the fungal
demonstrate greater efficacy compared to conventional strain, ultimately leading to its eradication. The genera-
pesticides (Fig. 9). This increased effectiveness could be tion of ROS under visible light illumination enabled the
attributed to improved absorption of the active ingre- disinfection of the fungus. Consequently, these TiO₂-
dients and higher bioavailability facilitated by NPs, NPs could be utilized for visible light-induced bacterial
consequently resulting in more efficient elimination of and fungal disinfection [248]. In another investigation,
infectious agents [315]. the antibacterial capabilities of TiO₂-NPs and TiO₂-NPs
TiO₂-NPs were employed to combat Spodoptera litto- doped with silver (Ag) and zinc (Zn) showed high anti-
ralis, a polyphagous pest that infests various crops like bacterial activity (in-vitro and greenhouse experiment)
cotton and vegetables such as tomatoes. An experiment against Xanthomonas perforans that causing tomatoes
tested six TiO₂-NP concentrations (31.25–1000 ppm) by spot disease at a concentration of 500–800 ppm under
feeding larvae leaves treated with these nanoparticles. visible light conditions. This enhanced activity was attrib-
Two weeks after application, mortality was assessed. The uted to the combinatorial effects of TiO₂ and the Ag and
findings unequivocally demonstrated the toxic effects of

Fig. 9 The advantages of utilizing TiO2-NPs as nanopesticides for enhancing and improving agricultural yield

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 26 of 40

Zn dopants [319]. Table 8 summarizes the activity of hairs across the root epidermis, moving symplastically
TiO2-NPs in agricultural sectors as nanopesticides. via plasmodesmata between cells, and subsequently
transported upwards through stem vascular tissue to
TiO2-NPs as nanofertilizers the leaves (Fig. 10).contributing to elevated chlorophyll
The incorporation of fertilizers can boost agricultural levels, augmented photosynthetic efficiency, and conse-
productivity. However, to address issues such as environ- quently enhanced plant biomass and yield (Fig. 11).
mental pollution and inefficient nutrient utilization, nano The choice of application technique hinges on the spe-
fertilizers emerge as a preferable option, potentially serv- cific crop, its development phase, and intended objec-
ing as a more effective and efficient alternative to conven- tives. Foliar spraying is prevalent for its direct delivery of
tional fertilizers. Additionally, nano fertilizers contribute nanoparticles to photosynthetic tissues, while soil appli-
to enhancing soil quality by mitigating the detrimental cation is favored for prolonged nutrient accessibility.
effects associated with the excessive application of con- The influence of TiO₂-NPs, a nano fertilizer, was stud-
ventional fertilizers [326]. TiO2-NPs can be utilized in ied on spinach seedlings during their growth and devel-
the context of nanofertilizers via diverse approaches, opment stages. The treatment with TiO₂-NPs resulted in
such as foliar administration, seed pretreatment involv- improved seed vigor and germination rates. Furthermore,
ing immersion, and soil hydroponic methods where at a 2.5% concentration of nanoTiO2, enhancements were
TiO2-NPs are incorporated into the irrigation solution observed in plant dry weight, Rubisco activity, chlo-
or combined with soil substrates [327]. Foliar spraying rophyll formation, and phytosynthetic rate during the
is a prevalent method for facilitating rapid absorption growth phase [328].
of TiO2-NPs through stomatal uptake and cuticle layer Mahmoodzadeh and coauthors investigate the effects
penetration. Their transport through cell walls and dis- of TiO2-NPs on the seedling vigor of canola plants [329].
tribution within the plant vascular system is facilitated The authors reported that the seedling and radical growth
by their size and surface traits. Conversely, soil applica- of canola was increased at high TiO2-NPs concentration
tions are frequently chosen due to its prolonged nutri- (2000 mg/L). In a related investigation, the plant growth
ent release capability. TiO2-NPs are internalized by root (vegetative parts, male and female flower appearance)

Table 8 Examples of TiO2-NPs used in the agricultural sector to control the infection with different pests, insects, and microbes
Nanoparticles Synthesized TiO2-NPs Target Host Results Refer-
treatment source ence
TiO₂-NPs Leaf aqueous extract of Aedes aegypti (larvae Not detected Maximum death of larvae and pupa was at- [318]
Pouteria campechiana and pupa) tained at 900 µg/ml
TiO₂-NPs Commercial with Tested against Cotton, tomatoes Toxic effects at all concentrations [316]
concentrations of Spodoptera littoralis
31.25–1000 ppm larvae
TiO₂-NPs + ZnO-NPs Chemical Bactericera cockerelli Tomato plants Causing 99% mortality under lab condi- [317]
TiO₂ (100 ppm), ZnO nymphs tions and 32% mortality under greenhouse
(250 ppm) conditions
TiO₂-NPs (co-doped Not detected Fusarium oxysporum Tomatoes Completely eradicated fungal strain under [248]
fluorine and nitrogen) visible-light condition
TiO₂-NPs + Ag/Zn Commercial with con- Xanthomonas Tomatoes High antibacterial activity under visible light [319]
dopants centration of 500–800 perforans
ppm
TiO₂-NPs Desmostachya bipinnata Spodoptera litura and Not detected The highest mortality of 96% and 94% against [320]
Aedes aegypti (larvae A. aegypti and S. litura respectively was at-
and pupa) tained at 900 µg/mL.
Solanum trilobatum pediculocidal, and A subpictus High mortality rate at [321]
larvicidal activities and Hyalomma 2–10 µg/mL of TiO2-NPs
anatolicum
Beauveria bassiana Noctuidae pests Helicoverpa armig- 50% mortality of caterpillars with lower [322]
era and Spodoptera toxicity
frugiperda
Moringa oleifera Bipolaris sorokiniana wheat 40 mg/L of TiO2-Np was sufficient to reduce [323]
infection disease severity
Bacillus thuringiensis Ephestia kuehniella Mediterranean Potential nano pesticides, 74% larval mortality [324]
larvae flour moth
Ag/Fe/TiO2-NPs Trichoderma harzianum Sclerotinia Anodontites Fungicidal action after 48 of TiO2-Np exposure [325]
sclerotiorum trapesialis

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 27 of 40

Fig. 10 Foliar spray and soil irrigation routes of TiO2-NPs as nanofertilizers

Fig. 11 Advantages of utilizing TiO2-NPs as nanofertilizers to improve plant growth traits

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 28 of 40

and phytosynthetic pigment content (chlorophyll, antho- sewage management, which interferes with the indis-
cyanins, and carotenoids) of Zea mays were affected by criminate use of pesticides and fertilizers in agriculture
spraying of TiO2-NPs at various TiO₂-NPs concentra- parties. Presently, the pollution of our environment is so
tions. The results showed that the 0.03% TiO₂-NPs con- widespread that it has turned into a critical issue. Water
centration had a notable effect on chlorophyll a and b, and soil are contaminated with harmful heavy metals,
carotenoids, total chlorophyll, and anthocyanins. Addi- chlorinated compounds, or dyes. At the same time, the
tionally, the pigmentation during the reproductive stage atmosphere is filled with noxious nitrogen oxides (NO),
was improved upon TiO₂-NPs spray as compared with carbon monoxide (CO), volatile organic compounds,
control. These activities could be due to the positive and chlorofluorocarbons (CFCs). Given the high levels of
impact of TiO₂-NPs on the electron transport chain and contamination, now is the time to employ advanced tech-
photosynthesis processes, thereby boosting pigment pro- nology for monitoring and identifying these pollutants in
duction [330]. our water and soil [339].
Recently, TiO2-NPs formed by fruit peel extract of Cit- Nanotechnology has experienced remarkable advance-
rus medica were used to improve the Capsicum annuum ments in the field of environmental protection over the
yield [331]. Also, the foliar spray of TiO2-NPs on sunflow- recent years [340]. Among its most promising contri-
ers at a concentration of 2.6 mg/L led to improved physi- butions are the significant applications in water and air
ological traits and nutritional parameters and increased remediation. The nano-size, surface area to volume ratio,
the content of oil [332]. Interestingly, the height of cori- chemical NPs stability, surface modifications, and shapes
ander plants, as well as their physiological traits, carot- are considered the unique features of NPs, making them
enoid contents, total sugars, amino acids, and phenols, superior activity for environmental applications either
were improved as a result of spraying with TiO2-NPs at in-situ or ex-situ [341]. Different types of available nano-
2, 4, and 6 ppm [333]. These studies collectively dem- materials and nanotools are being utilized to remediate
onstrate the potential of TiO₂-NPs to enhance various environmental contaminants [342]. Among the various
aspects of plant growth and development across varied materials, utilizing TiO₂-NPs as a remediating agent is
species, highlighting their promising applications in agri- witnessing a steady rise in applications such as water
culture (Table 9). purification, air cleaning, and soil decontamination. The
electronic band structure, high quantum efficiency, sta-
Environmental applications bility, and chemical inertness for TiO2-NPs enable them
Natural resources such as water, air, and soil are severely to be resilient and adaptable to diverse conditions and
affected worldwide, and their recovery is difficult due uses, especially for contaminant removal [343]. Table 10
to rapid population growth, urbanization, and indus- summarizes the activity of TiO2-NPs in removing soil
trialization. Several reasons lead to water and soil con- and water contaminants.
tamination. The main ones are the direct release of raw
industrial effluents without treatment into rivers and

Table 9 Summarizing the activity of TiO2-NPs in agricultural sectors as nanofertilizers

Synthesized by TiO₂-NPs Conc. Results Refer-
and type of treatment ence
Spinach seedlings 2.50%, seeds soaking Improved seed vigor, germination rates, plant dry weight, Rubisco activity, chloro- [328]
phyll formation, photosynthetic rate
Canola 2000 mg/L, seeds treatment Increased seedling and radical growth [329]
Zea mays 0.03%, spraying Notable effects on chlorophyll a and b, carotenoids, total chlorophyll, anthocya- [330]
nins, improved pigmentation during the reproductive stage
Capsicum annuum Not detected Enhanced plant growth [331]
Sunflowers 2.6 mg/L, foliar spray Improved physiological traits, nutritional parameters, increased oil content [332]
Coriander 2, 4, and 6 ppm, spraying Improved height, physiological traits, carotenoid contents, total sugars, amino [333]
acids, phenols
Wheat 25–100 µg/mL, foliar spray Reduced salinity and improved germination with 50 µg/mL TiO₂ -NPs [87]
Rice 1000 mg/L, root exposure Stress tolerance at lower conc. [334]
bell pepper 250 mg/L, leaves spray Increased disease resistance [335]
Rice 750 mg kg− 1, irrigation Boosted crop yield and improved rhizosphere enzymatic activity [336]
Moldavian balm 200 mg/L, added to nutrient Increased volatile oil content under salt conditions [337]
solution
Vitex plant 0-800 ppm, foliar spray Higher sugar content and boosted shoot and root dry weight mass [338]
Vitex agnus-castus L.

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 29 of 40

Table 10 Examples of different TiO2-NPs and their rule in soil and water remediation
Contaminants Treatment Results Reference
Diphenyl arsenic acid (DPAA) TiO2-NPs DPAA removal efficiency up to 82.7%; adsorption of [344]
inorganic arsenic byproducts
p-Nitrophenol TiO₂-NPs combined with pulsed discharge Removal of p-nitrophenol up to 88% within 10 min [345]
plasma
Cadmium (Cd) TiO₂-NPs combined with plant growth-pro- Enhanced Cd uptake, increased chlorophyll content, [346]
moting rhizobacteria (PGPR) and promoted Trifolium repens seedlings
Antimony (Sb) TiO₂-NPs combined with biochar Increased Sb accumulation and improve Sorghum [347]
bicolor seedlings
lead (Pb) TiO₂-NPs adsorbents (0.1 g) using surfactants 97% of pb(ii) ions removal from contaminated soil [348]
Cadmium (Cd) TiO₂-NPs synthesized by Trianthema portulacas- Efficient cd removal from industrial wastewater [349]
trum and Chenopodium quinoa
Cu²⁺ and Rhodamine B (Rh-B) TiO₂-NPs synthesized by Chlorella vulgaris High contaminants removal within 1 h. [350]
Industrial textile dyes, methylene TiO₂-NPs synthesized by durva herb High degradation activity of MB and Rh-B days after [351]
blue (MB) and Rh-B 50 min
Organic contaminants TiO₂/Arabic gum Degradation of ciprofloxacin and MB [352]
Phenols TiO2/Algae Complex 98% phenol degradation after nearly 19 h [353]
Methylene blue TiO₂-NPs synthesized by mulberry plant MB disintegration within 2 h [354]
Organic Dyes (RB19 &RR76) TiO₂-NPs synthesized by Eichhornia crassipes 0.08 g of TiO2-NPs in 60 min. under U.V at PH = 1 [355]
achieved complete degradation

Soil remediation remove up to 88% of p-nitrophenol within just 10 min

Soil contamination occurs due to the presence of hazard- [345].
ous compounds, particularly heavy metals at toxic lev- Also, a study examined the combined application of
els. The sources of soil pollution include manufacturing plant growth-promoting rhizobacteria (PGPR) and TiO2-
activities, mining operations, and landfill sites containing NPs for enhancing phytoremediation of cadmium (Cd)
industrial wastes like paint residues, electrical wastes, contaminated soil using Trifolium repens seedlings. The
batteries, and industrial or municipal sewage [356]. soil had 28% clay, 37% sand, 35% silt, 0.47% N, 7.1 mg/
Heavy metals pose a significant challenge as soil pollut- kg phosphorus, and pH 7.8. Different doses of TiO2-NPs
ants because they are non-degradable and persist in the and PGPR were applied separately and in combination
environment once introduced [357]. Numerous research to analyze their effects on Cd uptake, plant growth, and
studies have revealed intriguing findings pertaining to chlorophyll content. The combined application promoted
the use of TiO₂-NPs for soil remediation through the UV- plant growth, increased chlorophyll content, reduced the
facilitated degradation of organic contaminants present required TiO2-NPs dosage for phytoremediation, and
in the soil [358]. enhanced Cd uptake and T. repens growth in Cd-con-
The photodegradation of diphenyl arsenic acid (DPAA), taminated soil compared to individual applications. This
a pollutant resulting from leakage of arsenic weapons highlights the potential synergistic effects of PGPR and
with adverse health effects, has been studied using TiO2- TiO2-NPs for efficient phytoremediation of heavy metal-
NPs. Various operational parameters like NP dosage, polluted soils [346]. The combined application of biochar
radiation time, light intensity, and soil-water ratio were and TiO2-NPs was explored for phytoremediation of
optimized to enhance DPAA removal efficiency up to antimony (Sb) contaminated soil using Sorghum bicolor
82.7%. While TiO₂-NPs don’t completely convert DPAA, seedlings. The soil had a pH of 7.7, 1.12% N, 8.7 mg/kg
they adsorb its inorganic arsenic byproducts, playing a phosphorus, 28% clay, 37% sand, and 35% silt. Different
crucial role in mitigating DPAA contamination through concentrations of TiO2 and biochar were applied individ-
photocatalytic oxidation [344]. Another study investi- ually and in combination to assess their effects on plant
gated the photocatalytic degradation of p-nitrophenol, growth, Sb uptake and accumulation, and physiological
a bio-refractory and toxic organic compound widely responses in Sb-polluted soil. The results showed that
used in various industries, from contaminated soil using the combined biochar and TiO2-NPs treatment positively
TiO2-NPs combined with pulsed discharge plasma. The influenced plant growth and significantly increased Sb
mechanism involved removing p-nitrophenol from the accumulation compared to individual applications. The
soil and enhancing its degradation by increasing the findings demonstrated the potential of this approach for
pulsed discharge voltage. The authors proposed that the efficient phytoremediation of heavy metal-contaminated
pulsed discharge plasma could drive the photocatalysis of soils [347].
TiO2-NPs. The report revealed that this approach could

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 30 of 40

Water remediation Conclusion

Clean and fresh water is a vital necessity for daily life In summary, this review underscores the pivotal role of
and the life cycle of living organisms. Recently, water TiO2-NPs in nanotechnology. The principal findings
resources have been highly contaminated with a wide point out that TiO2-NPs showcase notable versatility
range of pollutants as a result of human activities [359]. and potential across diverse sectors, including biomedi-
Among these contaminants are physical (such as impu- cal research, agricultural advancement, and environmen-
rities that alter the water’s physical features), chemical tal restoration. Their distinctive characteristics, such as
(such as heavy metals and organic and inorganic pollut- high surface area and photoactivation customize them
ants), and biological pollutants. These pollutants have for various medical and ecological applications, including
negative and deleterious impacts on humans, animals, targeted drug delivery, photodynamic therapy, and envi-
and the ecosystem [360]. Therefore, the treatment of ronmental decontamination. The insights above empha-
wastewater based on nanoscience has been given more size the need for continued exploration of bio-based
attention. Adsorption and photocatalytic-based nano- synthetic techniques and detailed bioactivity mecha-
technology are among the approaches for wastewater nisms, focusing on enhancing their efficacy and safety
treatment that have gained a lot of traction due to their in healthcare applications and improving their usage in
eco-friendliness, sustainability, and cost-effectiveness. agronomic practices.
Additionally, they offer various other advantageous
properties that make them particularly suitable for tack- Future perspectives
ling the complex issue of water contamination [361]. The future of TiO2-NPs holds immense potential, with
In this regard, TiO2-NPs are widely employed in water numerous exciting avenues awaiting exploration. By
remediation (Fig. 12) due to their unique physical and addressing current challenges, leveraging cutting-edge
chemical features, highly biocompatible, robust oxida- technologies, and fostering interdisciplinary collabora-
tion efficiency, and unparalleled photocatalytic proper- tions, researchers can unlock the full potential of these
ties [10]. This prominence has led to a surge in research remarkable nanoparticles, driving innovation and
focused on harnessing the potential of TiO₂-NPs for advancing scientific frontiers across various domains.
wastewater treatment, with numerous studies exploring Synthesis Advancements: The green and biogenic syn-
their efficacy in this domain. thesis routes for TiO2-NPs have gained traction owing

Fig. 12 Enhanced photocatalytic activity of TiO2-NPs in organic pollutants degradation and their role in water remediation

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Ghareeb et al. Microbial Cell Factories (2024) 23:341 Page 31 of 40

to their eco-friendly nature and sustainability. However, environmental engineering. Such collaborative efforts
there is significant scope for optimization and scaling up foster cross-pollination of ideas, enable the exchange of
these biological synthesis methods to achieve industrial- knowledge and expertise, and accelerate the translation
scale production while maintaining monodispersity and of fundamental research into practical applications.
precise control over nanoparticle size, shape, and crys-
Acknowledgements
tallinity. Integrating cutting-edge techniques, such as The authors extend their appreciation to the Botany and Microbiology
machine learning and computational modeling, could Department, Faculty of Science, Al-Azhar University, Cairo, Egypt; Botany and
accelerate the design and development of novel biogenic Microbiology Department, Faculty of Science, Suez Canal University, Ismailia,
Egypt; and Microbiology and Immunology Department, Faculty of Medicine,
synthesis strategies, enabling the tailored production of Suez Canal University, Ismailia, Egypt, for their great cooperation and support
TiO2-NPs for specific applications. in achieving and publishing this research work.
Surface modifications and hybrid nanostructures: The
Author contributions
surface chemistry of TiO2-NPs plays a pivotal role in Rania M. Kishk; Conceptualization, validation, resources, visualization,
determining their functionality and performance. Future supervision, project administration; Ahmed Ghareeb, Amr Fouda, and Waleed
research endeavors could focus on developing innova- M. El Kazzaz; Conceptualization, methodology, software, validation, formal
analysis, investigation, data curation, writing—original draft preparation,
tive surface modification strategies, including doping, writing—review and editing, visualization, supervision, project administration.
functionalization, and incorporating biomolecules or All authors have read and agreed to the published version of the manuscript.”
polymers. These approaches can potentially enhance the
Funding
nanoparticles’ biocompatibility, stability, and targeted Open access funding provided by The Science, Technology & Innovation
delivery capabilities, expanding their applications in bio- Funding Authority (STDF) in cooperation with The Egyptian Knowledge Bank
medicine, sensing, and catalysis domains. (EKB).
This study received no external funding.
Moreover, integrating TiO2-NPs with other nanoma-
terials, such as graphene, carbon nanotubes, or quan- Data availability
tum dots, could give rise to hybrid nanostructures with No datasets were generated or analysed during the current study.

synergistic properties. These hybrid systems may exhibit

superior performance in areas like photocatalysis, energy Declarations
conversion, and optoelectronics, paving the way for Ethics approval and consent to participate
developing advanced technologies and devices. Not applicable.
Theragnostic applications: The remarkable photocata-
Consent for publication
lytic and optical properties of TiO2-NPs have enabled Not applicable.
their exploration in biomedical applications, including
cancer therapy, antimicrobial treatments, and bioimag- Competing interests
The authors declare no competing interests.
ing. Future research could delve into developing mul-
tifunctional TiO2-based nanoplatforms that seamlessly Received: 29 July 2024 / Accepted: 27 November 2024
integrate diagnostic and therapeutic capabilities. Such
theragnostic systems could facilitate early disease detec-
tion, targeted drug delivery, and real-time monitoring of
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