1.

Title Synthesis of metal oxide-based biopolymers and evaluation of their biomedical and
mechanical applications

2. Background and statement of the problem

Chitin is one of the most readily available biopolymers on Earth, its low solubility limits its uses
and applications. This biopolymer is made up of β-(1-4)-linked D-units that are dispersed at
random (Jiménez-Gómez et al., 2020). Chitosan has gained significant attention in recent years
due to its remarkable properties like high permeability, biodegradability, non-toxicity to humans,
and cost-effectiveness (Younes et al., 2014). The major sources of chitosan are shown below in
Table 1 (Islam et al., 2016; Kaur & Dhillon, 2014; Pellis et al., 2022):

Table 1. Sources of chitosan

Sources Percentage of Chitin (%)

Crustacean Exoskeleton 20-30
(Prawn)
Shrimp Cuticles (Shrimp) 30-40
Crab Cuticles (Crab) 15-30
Insects Cuticles (Beetles) 5-25
Fungi (Bread mold cell walls) 2-44

Chitin can be collected from a variety of sources but collecting from shrimp shells has greater
advantages due to its higher abundance, high chitosan content, consistency in quality, and
economic and environmental sustainability. Chitosan is most commonly used in many different
applications such as drug delivery systems, gene delivery, environmental applications, agricultural
applications, biosensors, antimicrobial coatings, antioxidant properties, wound dressings,
packaging materials etc. (Younes et al., 2014). Films made of chitosan biopolymer are robust and
challenging to break. Additionally, they have mechanical qualities similar to commercial polymers
made historically from petroleum-based derivatives and are flexible. They do not, however, have
a strong moisture barrier and are vulnerable to dampness. Several additional biopolymers can be
mixed with chitosan to create stable, mechanically compact films that have a lower water vapour
permeability (WVP) than the original polymer (Bonilla et al., 2012).

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Among the various polymers incorporated with nanoparticles, chitosan has drawn a lot of interest
because of its desirable characteristics (Naskar et al., 2019). Chitosan nanoparticles (NPs) have
enhanced antimicrobial and antioxidant potential because they combine the beneficial properties
of NPs (such as a large surface-to-volume ratio, high functionalization possibilities, and a better
capacity for drug delivery) with that of their chitosan foundation (Rashki et al., 2021). Chitosan
(CS)-based polymeric nanoparticles (NPs) have inherent advantageous characteristics such as
biocompatibility, biodegradability, non-toxicity, bioactivity, ease of synthesis, and targeted
specificity, which make them good drug carriers (Mikušová et al., 2021).

Numerous intriguing properties have been demonstrated for a range of metal oxide nanoparticle
types, including zinc oxide (ZnO), cerium oxide (CeO2), iron oxide (Fe2O3/Fe3O4), tin oxide
(SnO2), titanium dioxide (TiO2), and zirconium oxide (ZrO2). These nanoparticles are attractive
candidates because of their qualities, which include a high surface-to-volume ratio, good
adsorption capacity, high catalytic efficiency, and high surface reaction activity (Malhotra &
Kaushik, 2009).

Titanium dioxide (TiO2) nanoparticles (TNPs), a naturally occurring form of oxidized titanium,
have garnered significant attention among the various metal oxide nanomaterials due to their
exceptional antimicrobial and antioxidant properties (Santhoshkumar et al., 2014a), UV light
absorbance, hydrophobicity, and biocompatibility (Kominami et al., 1997; Mesgari et al., 2021).
It can also be used in drug delivery systems (Devanand Venkatasubbu et al., 2013).

Because of the synergistic antimicrobial effects of TNP and chitosan, researchers are interested in
studying chitosan-based TNP nanocomposites. Though chitosan and titanium dioxide
nanoparticles both show antioxidant properties individually, (Abd El-Hack et al., 2020;
Santhoshkumar et al., 2014b) there are good possibilities that they could show synergistic
antioxidant properties. TNPs have a photocatalytic function by releasing reactive oxygen species
(ROS) and chitosan, a cationic polymer that damages the membrane of bacteria (Babaei-Ghazvini
et al., 2021).

While there are a number of benefits that nanoparticles offer over traditional drug delivery
techniques, such as the removal of excessive doses, susceptibility across the intestinal epithelium,
restricted accessibility, first-pass effect, high reactivity, instability, and changes in plasma drug
levels, they are not without limits. The risk of nanotoxicity associated with growing NP uses is

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significant. The development of embryos, the central nervous system, the immunological system,
the respiratory system, the skin, the liver, the kidneys, and the reproductive system are all
negatively impacted by oxidative stress, which is strongly related to increased intracellular ROS
levels. The primary cause of these negative consequences is the size of NPs, which exhibit
different properties from larger counterparts with the same composition (Rout et al., 2018).

We will delve into the various synthesis techniques used to create chitosan-metal oxide
nanocomposites, the methods used to assess their morphological and structural characteristics, and
their applications in the biomedical and mechanical sectors.

By fostering research and innovation in this field, nations can strengthen their capabilities in
nanotechnology and materials science, positioning themselves at the forefront of advancements
with broad applications in healthcare, agriculture, and environmental protection. This not only
bolsters national health infrastructure but also promotes economic growth through the
development and commercialization of novel, high-value materials.

3. Literature Review

3.1. Mechanical Significance of titanium dioxide (TiO2) nanoparticles

Epoxy (EP) resins find extensive usage in a multitude of applications. They do not, however,
withstand crack initiation and propagation well and are brittle. The goal of current research has
been to employ inorganic nanoparticles to increase the fracture resistance of EP resins. TiO2
nanoparticle incorporation into the polymer matrix is one of the most researched methods. It is
possible for the resultant nanocomposites to have enhanced optical, electrical, mechanical,
rheological, and thermal properties. This study examines current research and developments in the
creation of polymer nanocomposites reinforced with TiO2 nanoparticles. Research indicates that
even at modest filler concentrations, the incorporation of TiO2 nanoparticles into EP resin can
provide significant mechanical characteristics (Pinto et al., 2015).

3.2. Significance of chitosan-based nanoparticles in drug delivery system

Nanoparticles play a crucial role in drug delivery and vaccine enhancement, offering non-invasive
administration routes. However, existing nanoparticle systems have limitations, often involving
harmful processes. Water-soluble polymers, particularly chitosan, provide a promising alternative.

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Chitosan stands out due to its biocompatibility, biodegradability, non-toxicity, and affordability.
Additionally, its positive charge and absorption-enhancing effect make it an attractive material for
drug delivery. This review emphasizes the widespread development of chitosan nanoparticles over
the past two decades, highlighting their simple preparation methods and applications in
pharmaceuticals. The focus includes chitosan nanoparticle technology, applications, and the
mechanisms of cell entry (Kumari et al., 2010; Tiyaboonchai, 2003).

3.3. Significance of chitosan-based titanium dioxide nanoparticle in

antimicrobial assay

Metal and metal oxide compounds have drawn interest due to their broad-spectrum antibacterial
activity, especially at the nanoscale. Metal oxide nanoparticles are the main topic of discussion,
with titanium dioxide (TiO2) being emphasized as a desirable semiconducting transition metal
oxide because of its affordability, non-toxicity, controllability, and resistance to chemical erosion.
The photocatalytic antibacterial activity of titanium dioxide nanoparticles, or TiO2 NPs, has been
the subject of much research due to its bio-related efficiency against bacterial contamination. TiO2
nanoparticles' antibacterial efficacy is closely associated with their morphological, structural, and
textural characteristics; crystalline structures especially anatase are crucial for achieving the best
possible physicochemical and antimicrobial qualities. The impact of synthesis parameters, such as
acid concentrations and hydrothermal temperatures, on the crystal structures and forms of TiO2
nanoparticles important factors that determine their antibacterial properties is examined. The focus
on the capacity of anatase structure to produce OH radicals in photocatalytic processes highlights
the possibility that it may influence the walls and membranes of bacteria, hence augmenting the
effectiveness of antimicrobial agents(Mareș et al., n.d.).

3.4. Antioxidative activities of chitosan derivatives

The investigated chitosan derivatives were shown to have a variety of antioxidative properties,
which varied according to the derivative molecule's sort, concentration, and DS with disaccharides.
Once these chitosan derivatives reached a specific concentration, the antioxidative activity
generally rose. Higher DS with disaccharide in chitosan derivatives was associated with a greater
scavenging effect on superoxide anion radicals, DPPH radicals, and H2O2. The disaccharide
chitosan derivatives were found to exhibit the best hydrogen peroxide scavenging performance

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among the several antioxidative activities investigated. All the chitosan derivatives examined
showed hydrogen peroxide scavenging ability of at least 60% at 400 μg/mL (Lin & Chou, 2004).

3.5. Biomedical application of pumpkin seeds

Pumpkin is a nutritious vegetable belonging to the Cucurbitaceae family. It is low in fat and an
excellent source of vitamins, minerals, dietary fibre, and β-carotene. Pumpkin seeds are considered
to be used for several health concerns including antidiabetic, antioxidant, anticarcinogenic, and
anti-inflammatory. It has also been applied to food product development. There are uses for almost
every part of the pumpkin plant that has to do with food or health. Pumpkin seeds have been used
as a taenicide, a diuretic, and a therapy for ascariasis, cestodiasis, and schistosomiasis. The seed
has anticancer, gynaecological, and hypolipidemic properties. Pumpkin seed oil has been utilized
as a nervine tonic and is a useful antioxidant due to its high phenolic content. Pumpkin seed extract
has antimicrobial properties. It is also used for the improvement of the haemoglobin content of
blood and to alleviate nausea and fever. This study has demonstrated various biomedical
applications of pumpkin seeds (Sharma et al., 2020).

3.6. Biosynthesis of ZnO NPs from pumpkin seeds and its significance against
cancer
Breast cancer which is a serious health hazard, requires new therapeutic alternatives. Zinc oxide
nanoparticles, or ZnO NPs, are one intriguing way that nanotechnology may be able to offer new
cancer treatments. Pumpkin seed extracts can be used to create ZnO NPs in a green approach, and
studies on human breast cancer cells have demonstrated their anticancer properties. The potential
of ZnO NPs as a novel breast cancer therapeutic option is demonstrated by this study (Tabrez et
al., 2022).

3.7. Chitosan-Zinc oxide nanocomposite

Zinc oxide nanoparticles (ZNPs) are cost-effective and possess unique properties such as catalytic,
electrical, optical, photostability, biocompatibility, and antimicrobial features. When exposed to
UV light, ZNPs generate electrons and holes, leading to the production of hydroxyl radicals and
superoxide anions. These radicals have potent oxidizing abilities, making ZNPs attractive for
various applications. However, creating well-dispersed ZnO/chitosan nanocomposites presents
challenges, as chitosan dissolves only in acidic conditions, while ZNPs transform into aqueous

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Zn2+ ions in acidity, forming chitosan-metal ion complexes rather than true nanocomposites. This
hurdle complicates the fabrication of optimal ZnO/chitosan nanocomposites (Pera-Titus et al.,
2004; Shahabi-Ghahfarrokhi & Babaei-Ghazvini, 2019).

3.8. Chitosan-Silver oxide nanocomposite

Silver nanoparticles (AgNPs), known for their antimicrobial properties, have been historically
recognized for their efficacy. Various antimicrobial mechanisms of AgNP activity include the
production and release of Ag+ ions, generation of reactive oxygen species (ROS), interference with
cell membranes, destabilization of ribosomes, and damage to mitochondrial and nucleic acids. The
synthesis of Ag-NPs can be achieved through physical, chemical, or biological methods. The
biological method is considered environmentally friendly, contrasting with physical and chemical
methods involving high energy and toxic solvents, limiting the potential biomedical and food
applications of the synthesized nanoparticles. In practical applications, chitosan-based films
incorporated with AgNPs have demonstrated enhanced antimicrobial effects against foodborne
pathogens, making them promising for active and intelligent food packaging solutions with added
antioxidant properties. These films, developed using a one-pot method, exhibit notable
antibacterial activity against both Gram-negative and Gram-positive bacteria (Abdallah et al.,
2020; Alavi & Kennedy, 2021; Chen et al., 2018; Nandana et al., 2022; Qin et al., 2019; Zhang &
Jiang, 2020).

3.9. Biopolymers as Food Packaging Materials

Chitosan biopolymer films made from shrimp shells are as flexible and strong as conventional
petroleum-based polymers, but they have a low moisture barrier and are sensitive to humidity,
which has limited their use in food-related applications. To address these limitations, scientists
have combined chitosan with other biopolymers, such as lipids and other polysaccharides, to create
stable and mechanically strong films with improved moisture barrier properties. Emulsion films
based on chitosan have shown promise in dried fruit and cereal applications. Advances in synthesis
techniques, such as the use of sophisticated catalytic systems, have also improved the
characteristics and durability of chitosan films, which previously had poor moisture resistance.
Harvard’s Wyss Institute has developed fully biodegradable biopolymers made from shrimp shells
that offer a greener alternative to traditional plastics. These chitosan-based materials can be used

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for a variety of applications, such as food packaging, bags, and degradable diapers, and they break
down quickly and release nutrients that are beneficial to plants. The study highlights the significant
benefits of using biopolymers, including the incorporation of nanoparticles with many
advantageous properties, such as heat and cold stability, impermeability to gases, strength,
firmness, and antibacterial capabilities. (Grujić et al., 2017).

4. Aim and objective

4.1. Aim

To investigate the effectiveness of metal oxide nanoparticles based on chitosan in the assessment
of mechanical and biomedical applications.

4.2. Objectives

• To collect chitosan from a local market and characterize its chemical and physical
properties.

• To synthesize metal oxide nanoparticles (e.g., zinc oxide, titanium dioxide, copper oxide
or iron oxide) with controlled size and shape for incorporation into the chitosan matrix.

• To conduct experiments to optimize the ratio of chitosan to metal oxide nanoparticles to

achieve the best chemical properties of the composite.

• To evaluate the antimicrobial, antioxidant, drug delivery system applications and

mechanical evaluation.

• To compare the performance of the chitosan-based nanoparticle composite with existing

materials in specific applications to highlight its advantages and limitations.

5. Research questions

• What conditions yield the most stable and effective titanium oxide nanoparticles using
TTIP (Titanium tertaisopropoxide) salt?
• What is the optimal composition ratio of titanium oxide nanoparticles, chitosan, and
phytochemicals for maximizing antimicrobial efficacy?

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 • How applicable is the synthesized composite for potential use in biomedical, mechanical,
or other relevant fields?
• What is the antimicrobial activity of the chitosan-phytochemical composite against a range
of microorganisms, including bacteria, fungi, and viruses?
• What are the potential challenges and future directions for further improving the
mechanical properties and applications of the composite?
• What are the environmental implications of using the chitosan-phytochemical composite
and TiO2 NPs as antimicrobial agents?

6. Methodology

• Phytochemical extraction: Household waste, pumpkin seeds will be washed with

deionized water and dried at room temperature in the dark for 28 days. Then the dried
leaves will be powdered entirely. Then 25 grams of fine powder of the seeds will be added
to 150 mL of distilled water and then stirred (5000rpm) at 60°C for 2 h. The mixture will
be cooled, filtered, and then stored in the refrigerator.

• Synthesis of metal oxide nanoparticles using phytochemicals: Titanium dioxide (TiO2)

nanoparticles and their composite will be synthesized using extracted phytochemicals by
the co-precipitation method (Kaushik et al., 2008).

• Incorporation with chitosan: Chitosan will be incorporated with the synthesized metal
oxide nanoparticles composite to form chitosan-based composite materials through a co-
precipitation process (Kaushik et al., 2008).

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 Fig.01 Flowchart of the synthesis process

• Characterisation: The composite materials will be extensively characterized using

techniques such as scanning electron microscopy (SEM), X-ray diffraction (XRD),
Fourier-transform infrared spectroscopy (FTIR), thermal analysis (TGA/DSC) and UV-Vis
Spectroscopy to understand the structural and thermal properties(Al-Alawy et al., 2018).

Chart.01 Characterization techniques

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 • Testing:

❖ Antimicrobial Testing: Disc diffusion method (Zaidan et al., 2005), Broth

microdilution assay (Balouiri et al., 2016), and time-kill assay (Balouiri et al., 2016)
❖ Antioxidant Testing: FRAP (ferric reducing antioxidant power) assay (Munteanu &
Apetrei, 2021), ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid))
radical scavenging assay (Munteanu & Apetrei, 2021).
❖ Drug Delivery Testing: In-vitro dissolution testing (Antolovich et al., 2002).
❖ Mechanical Testing: Tensile strength testing (Stevens & Poliks, 2003).

7. Time Frame/Activities Plan

Activities Time Period (2024)

Chitosan collection and characterization Month 1-2

Phytochemicals extraction Month 1-2

Synthesis of metal oxide nanoparticles Month 2-3

Composite formation and optimization Month 3-4

Characterisation of composites Month 3-4

Applications testing Month 4-5

Data analysis and thesis writing Month 5-6

8. Possible Outcomes

1. The project result will help to enhance the mechanical and functional properties of
chitosan-based composite incorporated with metal oxide nanoparticles.

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 2. Our findings will help to characterize and describe the structural properties of composite
materials.
3. The project result will evaluate the composite's potential applications in various industries,
including water treatment, biomedicine, and environmental remediation.
4. The individuals working on the project will have better communication, laboratory, and
academic abilities. Students at the master's level will have stronger research abilities.

9. Bibliography

Abd El-Hack, M. E., El-Saadony, M. T., Shafi, M. E., Zabermawi, N. M., Arif, M., Batiha, G. E.,
Khafaga, A. F., Abd El-Hakim, Y. M., & Al-Sagheer, A. A. (2020). Antimicrobial and
antioxidant properties of chitosan and its derivatives and their applications: A review.
International Journal of Biological Macromolecules, 164, 2726–2744.
https://doi.org/10.1016/J.IJBIOMAC.2020.08.153
Abdallah, O. M., EL-Baghdady, K. Z., Khalil, M. M. H., El Borhamy, M. I., & Meligi, G. A.
(2020). Antibacterial, antibiofilm and cytotoxic activities of biogenic polyvinyl alcohol-
silver and chitosan-silver nanocomposites. Journal of Polymer Research, 27(3), 1–9.
https://doi.org/10.1007/S10965-020-02050-3/FIGURES/4
Al-Alawy, A. F., Al-Abodi, E. E., & Kadhim, R. M. (2018). Synthesis and Characterization of
Magnetic Iron Oxide Nanoparticles by Co-Precipitation Method at Different Conditions.
Journal of Engineering, 24(10), 60–72. https://doi.org/10.31026/j.eng.2018.10.05
Alavi, M., & Kennedy, J. F. (2021). Recent advances of fabricated and modified Ag, Cu, CuO
and ZnO nanoparticles by herbal secondary metabolites, cellulose and pectin polymers for
antimicrobial applications. Cellulose, 28(6), 3297–3310. https://doi.org/10.1007/S10570-
021-03746-5/TABLES/2
Antolovich, M., Prenzler, P. D., Patsalides, E., McDonald, S., & Robards, K. (2002). Methods for
testing antioxidant activity. In Analyst (Vol. 127, Issue 1, pp. 183–198). Royal Society of
Chemistry. https://doi.org/10.1039/b009171p
Babaei-Ghazvini, A., Acharya, B., & Korber, D. R. (2021). Antimicrobial Biodegradable Food
Packaging Based on Chitosan and Metal/Metal-Oxide Bio-Nanocomposites: A Review.
Polymers 2021, Vol. 13, Page 2790, 13(16), 2790.
https://doi.org/10.3390/POLYM13162790
Balouiri, M., Sadiki, M., & Ibnsouda, S. K. (2016). Methods for in vitro evaluating antimicrobial
activity: A review. In Journal of Pharmaceutical Analysis (Vol. 6, Issue 2, pp. 71–79). Xi’an
Jiaotong University. https://doi.org/10.1016/j.jpha.2015.11.005

11 | P a g e
Bonilla, J., Atarés, L., Vargas, M., & Chiralt, A. (2012). Effect of essential oils and
homogenization conditions on properties of chitosan-based films. Food Hydrocolloids,
26(1), 9–16. https://doi.org/10.1016/J.FOODHYD.2011.03.015
Chen, M., Yan, J., Feng, Y., Duong, T.-T., Pham, L., Minh, T., Phan, J., & Park, A. (2018).
Antibacterial activity of poly (vinyl alcohol)—biogenic silver nanocomposite film for food
packaging material. Advances in Natural Sciences: Nanoscience and Nanotechnology, 9(2),
025020. https://doi.org/10.1088/2043-6254/AAC4EC
Devanand Venkatasubbu, G., Ramasamy, S., Ramakrishnan, V., & Kumar, J. (2013). Folate
targeted PEGylated titanium dioxide nanoparticles as a nanocarrier for targeted paclitaxel
drug delivery. Advanced Powder Technology, 24(6), 947–954.
https://doi.org/10.1016/J.APT.2013.01.008
Grujić, R., Vujadinović, D., & Savanović, D. (2017). Biopolymers as food packaging materials.
Advances in Applications of Industrial Biomaterials, 139–160. https://doi.org/10.1007/978-
3-319-62767-0_8/COVER
Islam, S., Bhuiyan, M. A. R., & Islam, M. N. (2016). Chitin and Chitosan: Structure, Properties
and Applications in Biomedical Engineering. Journal of Polymers and the Environment
2016 25:3, 25(3), 854–866. https://doi.org/10.1007/S10924-016-0865-5
Jiménez-Gómez, C. P., Cecilia, J. A., Guidotti, M., & Soengas, R. (2020). Chitosan: A Natural
Biopolymer with a Wide and Varied Range of Applications. Molecules 2020, Vol. 25, Page
3981, 25(17), 3981. https://doi.org/10.3390/MOLECULES25173981
Kaur, S., & Dhillon, G. S. (2014). The versatile biopolymer chitosan: potential sources,
evaluation of extraction methods and applications. Critical Reviews in Microbiology, 40(2),
155–175. https://doi.org/10.3109/1040841X.2013.770385
Kaushik, A., Khan, R., Solanki, P. R., Pandey, P., Alam, J., Ahmad, S., & Malhotra, B. D. (2008).
Iron oxide nanoparticles–chitosan composite based glucose biosensor. Biosensors and
Bioelectronics, 24(4), 676–683. https://doi.org/10.1016/J.BIOS.2008.06.032
Kominami, H., Kalo, J. I., Takada, Y., Doushi, Y., Ohtani, B., Nishimoto, S. I., Inoue, M., Inui,
T., & Kera, Y. (1997). Novel synthesis of microcrystalline titanium(IV) oxide having high
thermal stability and ultra-high photocatalytic activity: Thermal decomposition of
titanium(IV) alkoxide in organic solvents. Catalysis Letters, 46(3), 235–240.
https://doi.org/10.1023/A:1019022719479/METRICS
Kumari, A., Yadav, S. K., & Yadav, S. C. (2010). Biodegradable polymeric nanoparticles based
drug delivery systems. Colloids and Surfaces B: Biointerfaces, 75(1), 1–18.
https://doi.org/10.1016/J.COLSURFB.2009.09.001
Lin, H. Y., & Chou, C. C. (2004). Antioxidative activities of water-soluble disaccharide chitosan
derivatives. Food Research International, 37(9), 883–889.
https://doi.org/10.1016/J.FOODRES.2004.04.007

12 | P a g e
Malhotra, B. D., & Kaushik, A. (2009). Metal oxide–chitosan based nanocomposite for
cholesterol biosensor. Thin Solid Films, 518(2), 614–620.
https://doi.org/10.1016/J.TSF.2009.07.036
Mareș, M., Lim, S. E. H., Lai, K.-S., & Cristina, R.-T. (n.d.). Antimicrobial resistance : a one
health perspective. Retrieved November 17, 2023, from
https://books.google.com/books/about/Antimicrobial_Resistance.html?id=8pQtEAAAQBA
J
Mesgari, M., Aalami, A. H., & Sahebkar, A. (2021). Antimicrobial activities of chitosan/titanium
dioxide composites as a biological nanolayer for food preservation: A review. International
Journal of Biological Macromolecules, 176, 530–539.
https://doi.org/10.1016/J.IJBIOMAC.2021.02.099
Mikušová, V., Mikuš, P., & Piozzi, A. (2021). Molecular Sciences Advances in Chitosan-Based
Nanoparticles for Drug Delivery. https://doi.org/10.3390/ijms2217
Munteanu, I. G., & Apetrei, C. (2021). Analytical methods used in determining antioxidant
activity: A review. In International Journal of Molecular Sciences (Vol. 22, Issue 7). MDPI
AG. https://doi.org/10.3390/ijms22073380
Nandana, C. N., Christeena, M., & Bharathi, D. (2022). Synthesis and Characterization of
Chitosan/Silver Nanocomposite Using Rutin for Antibacterial, Antioxidant and
Photocatalytic Applications. Journal of Cluster Science, 33(1), 269–279.
https://doi.org/10.1007/S10876-020-01947-9/FIGURES/8
Naskar, S., Sharma, S., & Kuotsu, K. (2019). Chitosan-based nanoparticles: An overview of
biomedical applications and its preparation. Journal of Drug Delivery Science and
Technology, 49, 66–81. https://doi.org/10.1016/J.JDDST.2018.10.022
Pellis, A., Guebitz, G. M., & Nyanhongo, G. S. (2022). Chitosan: Sources, Processing and
Modification Techniques. Gels, 8(7). https://doi.org/10.3390/GELS8070393
Pera-Titus, M., García-Molina, V., Baños, M. A., Giménez, J., & Esplugas, S. (2004).
Degradation of chlorophenols by means of advanced oxidation processes: a general review.
Applied Catalysis B: Environmental, 47(4), 219–256.
https://doi.org/10.1016/J.APCATB.2003.09.010
Pinto, D., Bernardo, L., Amaro, A., & Lopes, S. (2015). Mechanical properties of epoxy
nanocomposites using titanium dioxide as reinforcement – A review. Construction and
Building Materials, 95, 506–524. https://doi.org/10.1016/J.CONBUILDMAT.2015.07.124
Qin, Y., Liu, Y., Yuan, L., Yong, H., & Liu, J. (2019). Preparation and characterization of
antioxidant, antimicrobial and pH-sensitive films based on chitosan, silver nanoparticles and
purple corn extract. Food Hydrocolloids, 96, 102–111.
https://doi.org/10.1016/J.FOODHYD.2019.05.017

13 | P a g e
Rashki, S., Asgarpour, K., Tarrahimofrad, H., Hashemipour, M., Ebrahimi, M. S., Fathizadeh, H.,
Khorshidi, A., Khan, H., Marzhoseyni, Z., Salavati-Niasari, M., & Mirzaei, H. (2021).
Chitosan-based nanoparticles against bacterial infections. Carbohydrate Polymers, 251,
117108. https://doi.org/10.1016/J.CARBPOL.2020.117108
Rout, G. K., Shin, H. S., Gouda, S., Sahoo, S., Das, G., Fraceto, L. F., & Patra, J. K. (2018).
Current advances in nanocarriers for biomedical research and their applications. Artificial
Cells, Nanomedicine, and Biotechnology, 46(sup2), 1053–1062.
https://doi.org/10.1080/21691401.2018.1478843
Santhoshkumar, T., Rahuman, A. A., Jayaseelan, C., Rajakumar, G., Marimuthu, S., Kirthi, A. V.,
Velayutham, K., Thomas, J., Venkatesan, J., & Kim, S. K. (2014a). Green synthesis of
titanium dioxide nanoparticles using Psidium guajava extract and its antibacterial and
antioxidant properties. Asian Pacific Journal of Tropical Medicine, 7(12), 968–976.
https://doi.org/10.1016/S1995-7645(14)60171-1
Santhoshkumar, T., Rahuman, A. A., Jayaseelan, C., Rajakumar, G., Marimuthu, S., Kirthi, A. V.,
Velayutham, K., Thomas, J., Venkatesan, J., & Kim, S. K. (2014b). Green synthesis of
titanium dioxide nanoparticles using Psidium guajava extract and its antibacterial and
antioxidant properties. Asian Pacific Journal of Tropical Medicine, 7(12), 968–976.
https://doi.org/10.1016/S1995-7645(14)60171-1
Shahabi-Ghahfarrokhi, I., & Babaei-Ghazvini, A. (2019). Using photo-modification to
compatibilize nano-ZnO in development of starch-kefiran-ZnO green nanocomposite as
food packaging material. International Journal of Biological Macromolecules, 124, 922–
930. https://doi.org/10.1016/J.IJBIOMAC.2018.11.241
Sharma, P., Kaur, G., Kehinde, B. A., Chhikara, N., Panghal, A., & Kaur, H. (2020).
Pharmacological and biomedical uses of extracts of pumpkin and its relatives and
applications in the food industry: a review. International Journal of Vegetable Science,
26(1), 79–95. https://doi.org/10.1080/19315260.2019.1606130
Stevens, E. S., & Poliks, M. D. (2003). Tensile Strength Measurements on Biopolymer Films.
Journal of Chemical Education, 80(7), 810–812. https://doi.org/10.1021/ED080P810
Tabrez, S., Khan, A. U., Hoque, M., Suhail, M., Khan, M. I., & Zughaibi, T. A. (2022).
Biosynthesis of ZnO NPs from pumpkin seeds’ extract and elucidation of its anticancer
potential against breast cancer. Nanotechnology Reviews, 11(1), 2714–2725.
https://doi.org/10.1515/NTREV-2022-0154/ASSET/GRAPHIC/J_NTREV-2022-
0154_FIG_012.JPG
Tiyaboonchai, W. (2003). Chitosan Nanoparticles : A Promising System for Drug Delivery. In
Naresuan University Journal (Vol. 11, Issue 3).
Younes, I., Hajji, S., Frachet, V., Rinaudo, M., Jellouli, K., & Nasri, M. (2014). Chitin extraction
from shrimp shell using enzymatic treatment. Antitumor, antioxidant and antimicrobial

14 | P a g e
 activities of chitosan. International Journal of Biological Macromolecules, 69, 489–498.
https://doi.org/10.1016/J.IJBIOMAC.2014.06.013
Zaidan, M. R. S., Rain, N., Badrul, A. R., Adlin, A., & Zakiah, &. (2005). In vitro screening of
five local medicinal plants for antibacterial activity using disc diffusion method. In Tropical
Biomedicine (Vol. 22, Issue 2).
Zhang, W., & Jiang, W. (2020). Antioxidant and antibacterial chitosan film with tea polyphenols-
mediated green synthesis silver nanoparticle via a novel one-pot method. International
Journal of Biological Macromolecules, 155, 1252–1261.
https://doi.org/10.1016/J.IJBIOMAC.2019.11.093

15 | P a g e