Synthesis, Characterization and Potential Drug Delivery Applications of Polymer-Coated Zinc Oxide Nanoparticles for Cancer-Targeted Therapy

Synthesis, Characterization and Potential Drug Delivery Applications of Polymer-Coated Zinc Oxide Nanoparticles for Cancer-Targeted Therapy | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Synthesis, Characterization and Potential Drug Delivery Applications of Polymer-Coated Zinc Oxide Nanoparticles for Cancer-Targeted Therapy L. O. Akinboyewa, A. F. Afolabi, M. A. Adekoya, O. I. Olusola, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7602894/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 22 You are reading this latest preprint version Abstract This study developed a chitosan-polyvinyl alcohol nanocomposite system (CH/PVA) incorporated with zinc oxide nanoparticles (ZnO) and conjugated with folic acid (FA) for targeted drug delivery applications. The synthesized materials CH/PVA, CH/PVA/ZnO, and CH/PVA/ZnO-FA were thoroughly characterized using multiple analytical techniques. Scanning electron microscopy (SEM) revealed that the folic acid-conjugated sample (CH/PVA/ZnO-FA) exhibited a distinct bead-like morphology, suggesting enhanced drug-loading capacity and therapeutic potential. Energy-dispersive X-ray spectroscopy (EDX) confirmed the elemental compositions (Zn, O, Mg, Na, C and Ca), showing prominent zinc signals (48.0 wt %) in CH/PVA/ZnO-FA, indicating successful nanoparticle incorporation and FA conjugation. X-ray diffraction (XRD) analysis demonstrated high crystallinity, with a strong diffraction peak at 28.5° corresponding to the (002) plane, reflecting optimal atomic packing density. Fourier-transform infrared spectroscopy (FTIR) shows functional groups of the samples including hydroxyl (-OH) and amine (-NH) stretching vibrations at 3440 cm⁻¹, confirming increase in hydrogen bond within the polymer matrix. Thermogravimetric analysis (TGA) revealed excellent thermal stability up to 350°C, suitable for biomedical applications. These comprehensive characterization results demonstrate that the CH/PVA/ZnO-FA nanocomposite possesses ideal structural, chemical, and thermal properties for use as an advanced nanocarrier system in targeted cancer therapy applications. Crab shells Ionic gelation Chitosan Nanoparticles Zinc oxide Polymer and Folic acid Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1. Introduction Nanotechnology is increasingly recognized for its role in advancing cancer treatment, particularly in improving targeted chemotherapy through polymeric nanoparticles as drug carriers. This polymeric material can be modified to deliver chemotherapeutic agents directly to cancerous cells while minimizing exposure to healthy tissues. Additionally, it can be designed to recognize specific grows on cancer cells, ensuring that the drug accumulates mainly at the tumor site, thereby increasing its efficacy (Zhao et al. , 2011). Nanoparticle drug delivery carrier have many advantages over small molecule therapeutics, including reducing side effects and increasing drug potency (Zhong et al., 2018 : Xu and Du. 2003). However, the synthesis of chitosan nanoparticles from available polymer chosen has ability to improve the timed released of drug molecules and transport small drug molecule into the desired location (Younes et al., 2012 ). A polymeric material would be use as a component for drug delivery system and it is originated from blends of synthetic and natural polymers coated with zinc oxide nanoparticles (ZnONPs), this is a new material that has much attention as a component for drug delivery system. Polymeric material consists of a chitosan blend with polyvinyl alcohol (CH/PVA) coated with zinc-oxide nanoparticles (CH/PVA/ZnO) and conjugate with folate receptor (CH/PVA/ZnO-FA). Chitosan is a biodegradable, biocompatible and antimicrobial polymer that is a natural carbohydrate. It can be synthesized by the partial N-deacetylation of chitin, a natural biopolymer derived from crustacean shells such as crabs, shrimps, and lobsters (Cheba 2011 : Dutta et al., 2004 ). Nanotechnology is prominent for its ability to facilitate targeted drug delivery to specific cells through polymeric nanoparticles. (Wu et al. , 2003; Tian et al 2014 ;Tiyaboochai 2003). Modifying zinc oxide nanoparticles (ZnO NPs) with chitosan has boosted metal-containing particles. (Tememe et al. , 2021; Ramasamy et al ., 2014; Tran et al ., 2014). Polymeric materials coated with ZnO NPs hold significant potential in drug delivery by acting as active agents, controlling drug release, and facilitating drug carrier to the affected portion (Tiyaboonchai, 2003 ). Additionally, integrating folic acid with polymeric materials further enhances their efficacy, making them unique and valuable for anticancer applications (Tsaih and Chen, 2003). Developing alternative drug carrier systems for targeting defective cells in the body is essential for advancing therapeutic precision. Chitosan nanoparticles can be synthesized using various methods such as ionotropic gelation, microemulsion, solvent diffusion, emulsification, polyelectrolyte, complexation, and inverse micelles (Divya and Jisha, 2011; Yusan et al. , 2024). Among these methods, the ionotropic gelation method, utilizing sodium tripolyphosphate (STPP) as a cross-linking agent is one of the best. This technique employs electrostatic interaction between positively charged chitosan and negatively charged TPP , forming stable polymeric nanoparticles. Such systems are increasingly studied for their multifunctionality in the pharmaceutical and food industries ( Somnuket al., 2011 ; Sireikhatim et al., 2015). This technique is not only enhancing the efficiency of nanoparticle production but also opens doors to new applications and advancements in various fields. It is widely preferred for its simplicity, non-toxic processing, mild reaction conditions, elimination of organic solvents, and capacity to achieve controlled release (Agnihotri et al., 2004 ). Expanding on these principles, this work investigates polymeric zinc oxide nanoparticles (ZnO NPs) conjugated with folic acid as a targeted drug delivery platform for anticancer applications. 2.0 Materials and Methods 2.1 Materials (750 g) of Crab shells were collected from Abata village, Akure South Local Government Area of Ondo State, Nigeria. The collected crab shells were thoroughly washed with distilled water to eliminate impurities. The samples were oven-drying at 70°C for 5 hours, then the oven-dried samples were mechanically grand into fine powder and sieved to achieve a uniform particle size of 105 µm. Chitosan isolation was performed through synthesis process including demineralization (acid treatment), deproteinization (alkaline hydrolysis), decolorization (organic solvent treatment), and deacetylation to enhance chitosan purity with little modification of Maher, et al ., 2008. The experimental reagents included acetic acid (solvent for chitosan dissolution), sodium tripolyphosphate (ionic crosslinker for chitosan nanoparticles), zinc acetate (precursor for ZnO nanoparticle synthesis), tocopherol acetate (model drug), polyvinyl alcohol (synthetic polymer for blend), glutaraldehyde (crosslinking agent), potassium hydroxide and sodium hydroxide (pH adjusters), and folic acid (targeting ligand). 2.2 Preparation of Polymeric material Chitosan nanoparticles with low molecular weight were synthesized via ionic gelation. A chitosan solution was prepared by dissolving 20 g of chitosan in 100 mL distilled water and 50 mL of 2% aqueous acetic acid (pH 5.4). To initiate crosslinking, 0.1% (w/v) sodium tripolyphosphate (STPP) solution gently added into chitosan solution with a constantly stirred using magnetic stirrer at room temperature. The colloidal suspension was centrifuged at 16,000 rpm for 30 minutes (40°C), and the resulting nanoparticles were lyophilized and washed with distilled water to ensure purity. In parallel, zinc oxide nanoparticles were prepared through pyrolysis. 50 g of zinc acetate dehydrate [Zn(CH 3 COO) 2 ·2H 2 O] was added to 25 g of tocopherol acetate together and heated at 70°C until it form homogenous solution. Then, the solution was heated to 120°C to evaporate residual solvents, yielding a viscous gel. Pyrolysis of the gel at 400°C for 5 hours in a muffle furnace produced crystalline ZnO nanoparticles. Chitosan nanoparticles (CHNPs 20 g) were dissolved in 100 mL of distilled water and 50 mL of 0.2 M acetic acid. The mixture was mixed with 50 mL of polyvinyl alcohol (PVA) and stirred for 5 minutes to form a homogeneous CH/PVA solution. To stabilize the composite, 4% glutaraldehyde was added as a crosslinking agent, followed by dropwise addition of potassium hydroxide solution (0.01 g in 10 mL H₂O) to form a homogenized system. 3.5% (w/v) zinc oxide nanoparticles (ZnO NPs) were added into the homogenized solution in a flask and kept in an oven for two days at room temperature. The samples obtained were washed with distilled water to remove undissolved glutaraldehyde at 25 0 C. Folic acid solution was obtained by dissolving 50ml of folic acid in 1M NaOH and was added to the homogenized solution by stirring at 45 0 C for 4 hours. Then, the final product CH/PVA/ZnO- FA was collected at room temperature. 3.0 Material Characterization 3.1 X-ray Diffraction (XRD) X-ray diffraction (XRD) : This technique is used to analysis the crystalline nature of the synthesized material. The wavelength of the incident x-ray is determined by Bragg’s law in Eq. 1 $$\:2dsin=n\lambda\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\left(1\right)$$ where, \(\:\:d\) denotes the distance between atomic planes (interplanar spacing), \(\:\theta\:\) is the angle of diffraction, \(\:\lambda\:\:\) represents wavelength, and \(\:n\) is an integer . The average size of crystalline index ( L ) was estimated using the Scherrer formula in agreement with (Gumukaya et al 2003, Popescus et al 2011). $$\:L\:=\:\frac{K\:X\:\lambda\:}{\beta\:\:X\:Sin\:\theta\:}\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\left(2\right)\:$$ where, \(\:K\) is a dimensionless shape factor (typically 0.91), \(\:\:\lambda\:\) is the wavelength of the incident X-rays (0.154 nm), \(\:\:\beta\:\) is the peak’s full width at half maximum (FWHM), and \(\:\:\theta\:\) is the Bragg angle. This calculation provides a means of assessing the nanocrystalline features of the sample. To quantify the degree of crystallinity in the material, the crystallinity sizes was determined using this expression. $$\:{C}_{r}I=\frac{{I}_{200}-{I}_{am}}{{I}_{200}}\times\:100\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\left(3\right)$$ where \(\:{I}_{200}\) is the maximum intensity of the lattice diffraction and \(\:{I}_{am}\) is the low intensity peak of amorphous region of the baseline at \(\:2\theta\:\) . 3.2 Scanning Electron Microscopy (SEM) A scanning electron microscope (SEM) determined the surface morphology of polymeric material. The image of the sample's surface at different areas or dimensions was determined using a scanning electron microscope (SEM). 3.3 Fourier Transform Infrared Spectroscopy (FTIR) Fourier transform infrared (FTIR) spectroscopy is a technique used to obtain the functional groups present in the composite material. An infrared spectrum of absorption or emission was performed with a range of 400 to 4000 cm⁻¹, which helps to confirm the presence of characteristic chemical bonds and interactions among the components. 4.0 Results and Discussions 4.1 Scanning Electron Microscopy of Polymeric Materials The SEM micrographs in Fig. 1 a, 1 b, and 1 c provide insight into the surface morphology and dispersion of zinc oxide nanoparticles (ZnO NPs) within various polymeric matrices, specifically chitosan/polyvinyl alcohol (CH/PVA), (CH/PVA/ZnO), and (CH/PVA/ZnO-FA) respectively. The image morphology of CH/PVA in Fig. 1 a reveals a spherical shape with a smooth loosely packed structures. This indicates an inhomogeneous surface typical of polymeric blends without nanofiller incorporation, this research is in supports of Cardo et al ., (2022). This structure may have impact in both the mechanical stability and drugs carrier if used in biomedical applications. The image surface CH/PVA/ZnO nanocomposite in Fig. 1 b, illustrates that the ZnO nanoparticles are dispersed throughout the polymer matrix, and enhancing surface uniformity. The particles appear more densely packed with increased surface roughness and micro-aggregates, which signifies successful incorporation of ZnO into the CH/PVA matrix. This improved morphology typically correlates with enhanced mechanical integrity, better thermal stability, and potentially higher drug-loading efficiency in agreement with Younes et al., ( 2012 ). Figure 1 c reveals further structural refinement. The surface becomes even more compact, with fewer visible and well-dispersed nanoparticles forming a consistent, densely packed layer. The folic acid conjugation likely promotes additional interactions within the matrix, reducing agglomeration and enhancing dispersion of ZnO nanoparticles. This modification is especially beneficial in targeted drug delivery, as folic acid improves cellular uptake by targeting folate receptors commonly over expressed in cancer cells. This result is in agreement with Tememel et al. , (2012) 4.2 Energy dispersive X-ray spectroscopy of Polymeric Materials The Energy-Dispersive X-ray (EDX) spectroscopy analysis reveals discrete elemental compositions across the three polymer-nanoparticle formulations designed for targeted cancer drug delivery. The first spectrum in Fig. 3 a represents CH/PVA, which shows a predominant calcium content (45.0%), along with significant amounts of oxygen (25.0%), magnesium (15.3%), carbon (9.2%), phosphorus (3.5%), and sodium (2.0%). This baseline polymer blend exhibits no zinc content, providing a clear compositional reference point for the subsequent functionalized formulations, this report is in line with Agil-Abraham et al. , (2016). The second spectrum in Fig. 3 b depicts CH/PVA/ZnO, which demonstrates the successful incorporation of zinc oxide nanoparticles, with zinc appearing prominently (30.0%) while maintaining calcium as a major component (38.0%). The relative proportions of other elements are adjusted accordingly with oxygen (15.0%), carbon (6.2%), magnesium (5.3%), phosphorus (3.5%), and sodium (2.0%). The introduction of zinc oxide nanoparticles significantly alters the elemental profile compared to the plain polymer blend, confirming effective nanoparticle integration within the polymeric matrix. Similar research by Tememel et al ., (2021) also revealed these properties. The third in spectrum in Fig. 3 c corresponds to CH/PVA/ZnO-FA, which displays the highest zinc concentration (48.0%), indicating further enhanced zinc oxide nanoparticle loading after folic acid conjugation. This formulation shows a notable shift in calcium content (reduced to 15.3%) while maintaining substantial oxygen (20.0%) and carbon (10.2%) with lesser amounts of magnesium (4.5%) and sodium (2.0%), this result is in line with Tiyaboonchai, ( 2003 ). The elevated zinc concentration in this final formulation suggests that the addition of folic acid may facilitate greater nanoparticle encapsulation or surface attachment, potentially enhancing the system's cancer-targeting capabilities through folate receptor-mediated mechanisms. This finding is consistent with Shanmuga et al ., (2013). The progressive increase in zinc content from CH/PVA to CH/PVA/ZnO-FA corresponds with the intended design of a drug delivery system with incrementally enhanced therapeutic potential for targeting cancer cells is in supports of Umesh et al .,( 2011) article. (c) CH/PVA/ZnO –FA 4.3 X-ray Diffraction Analysis of Polymeric Materials Figure 3 a displays characteristic crystalline peaks at 10.0°, 27.0°, 28.0°, 46.0°, 50.0°, and 69.0°, consistent with the crystalline phases of chitin and chitosan derived from crab shells, as well as the semi-crystalline nature of polyvinyl alcohol (PVA). These peaks correspond to the crystallographic planes (002), (100), (101), (200), (220), and (310), respectively, confirming the crystalline structure of the CH/PVA blend. The most intense peak at 27.0° (101) indicates a semi-crystalline polymer matrix, attributed to hydrogen bond between the two samples molecules. This report is in accord with Hang et al., ( 2010 ); Nadia et al ., (2018). The XRD pattern of CH/PVA/ZnO exhibits distinct peaks at 10.0°, 27.5°, 28.0°, 30.0°, 40.0°, 60.0°, and 65.5°in Fig. 3 b, assigned to the lattice planes (002), (100), (101), (200), (220), (300), and (311), respectively. The prominent peak at 28.0° (101) confirms the crystalline structure of ZnO nanoparticles embedded within the polymeric matrix. The presence of these peaks suggests structural modification due to the interaction between chitosan, PVA, and ZnO, leading to the formation of new crystalline phases or polymorphs. Similar research conducted by Casettari et al ., (2013) ; Gocho et al ., (2000); Kardas et al ., (2012) revealed same properties. The XRD pattern of CH/PVA/ZnO-FA (Fig. 3 c) reveals peaks at 10.0°, 26.0°, 28.5°, 30.0°, 40.0°, 50.0°, and 70.0°, corresponding to the planes (100), (002), (101), (200), (220), (300), and (311). The most intense peak at 28.5° (002) signifies enhanced crystallinity and uniform atomic stacking along the c-axis. This preferred orientation indicates improved crystalline alignment, which influence material properties such as mechanical strength and thermal stability. However, variations in peak intensity may also introduce micro strain or lattice defects due to atomic vacancies, potentially reducing crystallite size. This result is in agreement with (Nagarwal et al ., 2012; Zaman et al ., 202; D-campose et al ., 2004; Xie et al ., 2007; Herdiana et al ., 2021). Table 4.1 indicates that the crystallinity index of CH/PVA/ZnO-FA is 78%, which is higher than CH/PVA and CH/PVA/ZnO. Other parameters, including crystallite size (L), d-spacing (d) and full width at half maximum (FWHM). The increased crystallinity index of CH/PVA/ZnO-FA significantly enhances its physical, mechanical, and thermal properties of the samples and offers potential for a more sustained release profile, making it a promising polymer for carcinogenic applications. Table 4.1 Crystallographic Parameters Obtained from XRD Patterns S/N Sample 2θ ( 0 ) d (nm) L (nm) FWHM \(\:{C}_{r\:}\) I (%) 1 2 3 CH/PVA CH/PVA/ZnO CH/PVA/ZnO–FA 27.0 28.0 28.2 0.66 1.71 2.65 0.35 0.28 0.27 0.50 0.51 0.52 72 74 78 4.4 Fourier Transform (FTIR) Spectral Analysis of Polymeric Materials FTIR spectroscopic analysis of the synthesized polymeric materials in Figs. 3 , show their molecular architecture and intermolecular interactions that illustrate the drug delivery functionality. The absorption band of the CH/PVA polymer blend is shown in Fig. 3 . The broad absorption band at 3438 cm⁻¹ corresponds to the stretching vibrations of hydroxyl (O-H) and amine (N-H) groups respectively. This peak's relatively sharp profile suggests increased polymer chain interactions may enhance drug carrier capacity. The peak values observed at 2926 cm⁻¹ and 2400 cm⁻¹ confirm the presence of alkyl (C-H) stretching, whereas those between 1775–1131 cm⁻¹ indicate the presence of carbonyl (C = O) groups from acetate. The observed minor peak shifts compared to bulk materials indicate modified molecular interactions resulting from nanoscale particle formation, which could optimize drug loading efficiency, This result seen is agreement with similar studies by Ndamitso et al.,( 2020); Zhong et al., ( 2018 ). The CH/PVA/ZnO composite exhibits intensified absorption features. The broad absorption band of O-H/N-H stretching vibrations at 3440 cm⁻¹ shows increased intensity and a transmittance level of 38%, suggesting enhanced hydrogen bonding. The amide I band shift to 1633 cm⁻¹ reflects structural modifications from ZnO incorporation. The maintained peaks at 2926 cm⁻¹ (C-H) and 1039 cm⁻¹ (C-O) verify retained glutaraldehyde crosslinks. These spectral changes indicate a strong intermolecular interactions that may improve drug encapsulation stability, this aligns with findings from Sumaila et al.,( 2020); Mohammed et al ., (2019). Figure 3 reveals the FTIR profile of the FA-conjugated system. The broad absorption band of 3442cm⁻¹ confirms the presence of persistent strong hydrogen bond which is crucial for drug-polymer interactions. Key features include the amide I band, C-H, and C-O band reveal modified glycosidic linkages. These spectral modifications confirm successful F-A functionalization while maintaining the polymer's structural integrity, essential for targeted drug delivery, similar research by Mohammad-pourd, et al.,( 2010);Tolaimate et al., ( 2003 ) revealed same result. 4.5 Thermogravimetric analysis of Polymeric Material The TGA curves for CH/PVA, CH/PVA/ZnO, and CH/PVA/ZnO-FA reveal distinct thermal behaviors that reflect the influence of ZnO nanoparticles and folic acid (FA) on the polymer matrix. For the CH/PVA sample (Fig. 6 a), the initial weight loss observed below 100°C is minimal and corresponds to the evaporation of adsorbed moisture, as indicated by weight retention of 99.7% at 96°C. The main decomposition event occurs between approximately 300°C and 400°C, where the polymer undergoes rapid weight loss. The sharp decline in the temperature observed signifies the breakdown of the chitosan/PVA backbone, with a major decomposition temperature around 375°C, where 70.5% of the mass is lost. This report is in agreement with Agnihotri et al., ( 2004 ). In addition, incorporating ZnO nanoparticles into the CH/PVA matrix, as shown in the CH/PVA/ZnO sample (Fig. 6 b), slightly enhances the thermal stability of the composite, the onset of decomposition remains in the same temperature range, but the weight loss 72.2% occurs at 382°C, indicating a marginal increase in thermal resistance. The residue at 450°C remains at 10.75%, suggesting that ZnO nanoparticles, being inorganic, contribute to the non-volatile residue. The improved thermal stability can be attributed to the strong interfacial interactions between the polymer chains and ZnO nanoparticles, which restrict the mobility of the polymer chains and delay thermal degradation. This enhancement is particularly beneficial for drug delivery applications, as it ensures that the nanocomposite carrier can withstand higher processing and physiological temperatures without premature degradation. This result is similar to Somnuk et al., ( 2011 ); Tian et al.,( 2014) reports The CH/PVA/ZnO-FA sample (Fig. 6 c) reveals a slightly different thermal profile. The major decomposition temperature observed at 350°C and 70.5% weight loss are lower than the CH/PVA/ZnO sample. This slight reduction in thermal stability may be due to the introduction of folic acid, which, enhancing the targeting capability of the nanocomposite for cancer cells. Moreover, the residue at 450°C remains consistent at 10.75%, indicating that the inorganic content (ZnO) still dominates the final residue. The presence of folic acid introduced has a significant functional advantage for drug delivery system, even if it slightly compromises thermal stability, this finding is in agreement with Gupta and Kompella (2010); Cheba, ( 2011 ); Ali et al., ( 2019 ) reports. Conclusion The comprehensive characterization of CH/PVA, CH/PVA/ZnO, and CH/PVA/ZnO-FA nanocomposites reveals a systematic enhancement of structural, elemental, crystallographic, molecular, and thermal properties tailored for anticancer drug delivery. SEM analysis illustrates progressive morphological refinement from the porous CH/PVA matrix to the optimally dispersed FA-conjugated system, where improved nanoparticle distribution and reduced defects enhance drug-loading capacity and release kinetics. Complementary EDX results confirm the evolution through increasing zinc content (0% → 30% → 48%), validating successful ZnO incorporation and FA-mediated nanoparticle enrichment while maintaining polymer backbone integrity through consistent oxygen/carbon signatures. XRD patterns corroborate these findings, showing enhanced crystallinity (peaking at 43.3% for CH/PVA/ZnO-FA) and atomic ordering through distinct diffraction planes, particularly the intensified 28.5° (002) peak indicating structural perfection. FTIR spectroscopy reveals preserved hydrogen bonding networks and new vibrational modes, confirming molecular stability while demonstrating strengthened polymer-nanoparticle interactions and successful FA functionalization. Thermal analysis completes this picture, showing ZnO's stabilizing effect (382°C decomposition temperature) and FA's strategic trade-off between modest thermal reduction (350°C) and gained targeting capability. Together, these characterizations prove the nanocomposites achieve an optimal balance: morphological uniformity for drug loading, elemental specificity for therapeutic action, crystallographic stability for mechanical strength, molecular precision for controlled release, and thermal resilience for processing - all while incorporating cancer-targeting functionality. The FA-conjugated system emerges as particularly promising, combining structural advantages with biological targeting through folate receptors. This multidisciplinary validation confirms the nanocomposites' potential as advanced, multifunctional platforms for targeted carcinogenic disease treatment, where material properties align precisely with therapeutic requirements Declarations 2 Funding Declaration This research did not receive any specific funding or grant from institution – based or non-governmental organization. 3 Clinical Trial Number : Not applicable 4 Consent to Publish Declaration : Not applicable 5 Ethics and Consent to Participate Declarations : Not applicable 6 Authors Contributions L.O.A: Conceptualization, investigation, Visualization, Writing – Original Draft Preparation A.F.A: Formal Analysis, Writing-Reviewing and Editing. M.A.A: Methodology, Draft Reviewing, Editing and Discussion O.I.O: Methodology, Formal Analysis, Draft Reviewing, and Supervision. S.S.O : Project Administration, Methodology, Validation and Supervision. 7 Competing interest Declaration : No completing interest exists in this work. 8 Author Email Discrepancy : [email protected] 9 City in Affiliation : b Department of Physics, Wigwe University, Isiokpo, Rivers State, Nigeria. 10 Acknowledgements: I sincerely wish to express my profound gratitude to the department of Physics, Federal University of Technology, Akure, for their valuable contribution during the material preparation. My special thanks go to Dr. Francis Alo of the Department of Material science and Engineering, Obafemi Awolowo University, Ile –Ife, Nigeria, for his immense support during the material characterization. Decision: Minor revision 1 Data Availability Statement : All data generated or analysed during this study are included in this published article. 2 Permissions to collect the crab shells: Crab shells are available in area surrounding our local river, therefore, we don’t need licenses for collection of shells. 3 Article Type: Thanks for the observation. This a pure research works. Decision: Minor revision on 13 oct 2025 1 Revise title: Synthesis, Characterization and Potential Drug Delivery Applications of Polymer-Coated Zinc Oxide Nanoparticles for Cancer-Targeted Therapy 2 Data availability statement : The submission system under the 'Declaration' section by following step: Done References Abhinay S, Kumar D, Meenakshi A Dua., Gursharan, Singh, Chhatwal and AtulKuma, Johr. (2012). 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Zhong Z, Xing R, Liu S, Wang L, Cai S, Li P (2008) Synthesis of acyl thiourea derivatives of chitosan and their antimicrobial activities in vitro. Journal of Carbohydrate Resource 343:566 Zhong Z, Chen R, Xing R, Chen X, Liu S, Guo Z, Ji X (2018) Synthesis and antifungal properties of sulfanilamide derivatives of chitosan. Elsevier Ltd. All rights reserved Cardoso G, Sirlene MM, Pereira W, Lopes MA (2022) Composition Effects on the MorphologyofPVA/Chitosan Electrospun Nanofibers . 14 (22), 4856–4856. doi.org/10.3390/polym14224 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 18 Mar, 2026 Reviews received at journal 07 Jan, 2026 Reviews received at journal 03 Jan, 2026 Reviews received at journal 03 Jan, 2026 Reviewers agreed at journal 29 Dec, 2025 Reviewers agreed at journal 28 Dec, 2025 Reviews received at journal 27 Dec, 2025 Reviews received at journal 27 Dec, 2025 Reviewers agreed at journal 26 Dec, 2025 Reviewers agreed at journal 25 Dec, 2025 Reviewers agreed at journal 25 Dec, 2025 Reviews received at journal 01 Dec, 2025 Reviewers agreed at journal 22 Nov, 2025 Reviewers agreed at journal 21 Nov, 2025 Reviewers agreed at journal 21 Nov, 2025 Reviews received at journal 19 Nov, 2025 Reviewers agreed at journal 10 Nov, 2025 Reviewers agreed at journal 10 Nov, 2025 Reviewers invited by journal 09 Nov, 2025 Editor assigned by journal 14 Oct, 2025 Submission checks completed at journal 11 Oct, 2025 First submitted to journal 11 Oct, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7602894","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":535109768,"identity":"3b7b9ca1-f1f0-46e0-853a-9d76ce54f788","order_by":0,"name":"L. O. 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1","display":"","copyAsset":false,"role":"figure","size":112678,"visible":true,"origin":"","legend":"\u003cp\u003eSchematic diagram of Experimental Procedure of Polymeric Materials\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7602894/v1/f189ad637b5a11288ad2282c.png"},{"id":94992580,"identity":"fbe81b84-2e72-46bf-858a-35ff0bafaa2e","added_by":"auto","created_at":"2025-11-03 07:21:48","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":427732,"visible":true,"origin":"","legend":"\u003cp\u003eSEM image of (a) CH/PVA, (b) CH/PVA/ZnO, and (c) CH/PVA/ZnO-FA.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7602894/v1/bd1178a65e263a06194fd82b.png"},{"id":94992593,"identity":"aed60b08-b22e-46a2-8124-787fbf6d6076","added_by":"auto","created_at":"2025-11-03 07:21:51","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":374383,"visible":true,"origin":"","legend":"\u003cp\u003eEnergy dispersive X-ray spectroscopy (EDX) (a) CH/PVA, (b) CH/PVA/ZnO and\u003c/p\u003e\n\u003cp\u003e(c) CH/PVA/ZnO –FA\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7602894/v1/f090d610f7fcf342ea76b624.png"},{"id":94992597,"identity":"f38115eb-3995-49a7-9c80-2d9fe99e070b","added_by":"auto","created_at":"2025-11-03 07:21:51","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":250538,"visible":true,"origin":"","legend":"\u003cp\u003eX-ray diffraction patterns (XRD), (a) CH/PVA, (b) CH/PVA/ZnO (c) CH/PVA/ZnO –FA\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-7602894/v1/166ed057e90ab1edbd1503ee.png"},{"id":94992572,"identity":"54cc5d31-7c1e-42a6-b0c7-ffd3a3f4108f","added_by":"auto","created_at":"2025-11-03 07:21:47","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":259178,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFourier transform infrared (FTIR) spectra for Polymeric Material\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-7602894/v1/9b1d065b00f2ae08b782f520.png"},{"id":95000863,"identity":"26da487c-9fd1-4118-b42f-339d86f7cd97","added_by":"auto","created_at":"2025-11-03 09:00:32","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":516608,"visible":true,"origin":"","legend":"\u003cp\u003eThermogravimetric analysis ( TGA) of(a) CH/PVA, (b) CH/PVA/Zn (c) CH/PVA/ZnO –FA\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-7602894/v1/880b91f6dc7fcb1cf0dda891.png"},{"id":95001658,"identity":"12400391-4753-4372-bc83-fd128319a42b","added_by":"auto","created_at":"2025-11-03 09:02:47","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2846244,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7602894/v1/09652966-2c24-4415-9029-727733219668.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Synthesis, Characterization and Potential Drug Delivery Applications of Polymer-Coated Zinc Oxide Nanoparticles for Cancer-Targeted Therapy","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eNanotechnology is increasingly recognized for its role in advancing cancer treatment, particularly in improving targeted chemotherapy through polymeric nanoparticles as drug carriers. This polymeric material can be modified to deliver chemotherapeutic agents directly to cancerous cells while minimizing exposure to healthy tissues. Additionally, it can be designed to recognize specific grows on cancer cells, ensuring that the drug accumulates mainly at the tumor site, thereby increasing its efficacy (Zhao \u003cem\u003eet al.\u003c/em\u003e, 2011). Nanoparticle drug delivery carrier have many advantages over small molecule therapeutics, including reducing side effects and increasing drug potency (Zhong et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e: Xu and Du. 2003). However, the synthesis of chitosan nanoparticles from available polymer chosen has ability to improve the timed released of drug molecules and transport small drug molecule into the desired location (Younes et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). A polymeric material would be use as a component for drug delivery system and it is originated from blends of synthetic and natural polymers coated with zinc oxide nanoparticles (ZnONPs), this is a new material that has much attention as a component for drug delivery system. Polymeric material consists of a chitosan blend with polyvinyl alcohol (CH/PVA) coated with zinc-oxide nanoparticles (CH/PVA/ZnO) and conjugate with folate receptor (CH/PVA/ZnO-FA). Chitosan is a biodegradable, biocompatible and antimicrobial polymer that is a natural carbohydrate. It can be synthesized by the partial N-deacetylation of chitin, a natural biopolymer derived from crustacean shells such as crabs, shrimps, and lobsters (Cheba \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2011\u003c/span\u003e: Dutta et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). Nanotechnology is prominent for its ability to facilitate targeted drug delivery to specific cells through polymeric nanoparticles. (Wu \u003cem\u003eet al.\u003c/em\u003e, 2003; Tian et al \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2014\u003c/span\u003e;Tiyaboochai 2003).\u003c/p\u003e\u003cp\u003eModifying zinc oxide nanoparticles (ZnO NPs) with chitosan has boosted metal-containing particles. (Tememe \u003cem\u003eet al.\u003c/em\u003e, 2021; Ramasamy \u003cem\u003eet al\u003c/em\u003e., 2014; Tran \u003cem\u003eet al\u003c/em\u003e., 2014). Polymeric materials coated with ZnO NPs hold significant potential in drug delivery by acting as active agents, controlling drug release, and facilitating drug carrier to the affected portion (Tiyaboonchai, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). Additionally, integrating folic acid with polymeric materials further enhances their efficacy, making them unique and valuable for anticancer applications (Tsaih and Chen, 2003). Developing alternative drug carrier systems for targeting defective cells in the body is essential for advancing therapeutic precision. Chitosan nanoparticles can be synthesized using various methods such as ionotropic gelation, microemulsion, solvent diffusion, emulsification, polyelectrolyte, complexation, and inverse micelles (Divya and Jisha, 2011; Yusan \u003cem\u003eet al.\u003c/em\u003e, 2024). Among these methods, the ionotropic gelation method, utilizing sodium tripolyphosphate (STPP) as a cross-linking agent is one of the best. This technique employs electrostatic interaction between positively charged chitosan and negatively charged \u003cem\u003eTPP\u003c/em\u003e, forming stable polymeric nanoparticles. Such systems are increasingly studied for their multifunctionality in the pharmaceutical and food industries \u003cb\u003e(\u003c/b\u003eSomnuket al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; \u003cem\u003eSireikhatim et al., 2015).\u003c/em\u003e This technique is not only enhancing the efficiency of nanoparticle production but also opens doors to new applications and advancements in various fields. It is widely preferred for its simplicity, non-toxic processing, mild reaction conditions, elimination of organic solvents, and capacity to achieve controlled release (Agnihotri et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). Expanding on these principles, this work investigates polymeric zinc oxide nanoparticles (ZnO NPs) conjugated with folic acid as a targeted drug delivery platform for anticancer applications.\u003c/p\u003e"},{"header":"2.0 Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1 Materials\u003c/h2\u003e\u003cp\u003e(750 g) of Crab shells were collected from Abata village, Akure South Local Government Area of Ondo State, Nigeria. The collected crab shells were thoroughly washed with distilled water to eliminate impurities. The samples were oven-drying at 70\u0026deg;C for 5 hours, then the oven-dried samples were mechanically grand into fine powder and sieved to achieve a uniform particle size of 105 \u0026micro;m. Chitosan isolation was performed through synthesis process including demineralization (acid treatment), deproteinization (alkaline hydrolysis), decolorization (organic solvent treatment), and deacetylation to enhance chitosan purity with little modification of Maher, \u003cem\u003eet al\u003c/em\u003e., 2008.\u003c/p\u003e\u003cp\u003eThe experimental reagents included acetic acid (solvent for chitosan dissolution), sodium tripolyphosphate (ionic crosslinker for chitosan nanoparticles), zinc acetate (precursor for ZnO nanoparticle synthesis), tocopherol acetate (model drug), polyvinyl alcohol (synthetic polymer for blend), glutaraldehyde (crosslinking agent), potassium hydroxide and sodium hydroxide (pH adjusters), and folic acid (targeting ligand).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2 Preparation of Polymeric material\u003c/h2\u003e\u003cp\u003eChitosan nanoparticles with low molecular weight were synthesized via ionic gelation. A chitosan solution was prepared by dissolving 20 g of chitosan in 100 mL distilled water and 50 mL of 2% aqueous acetic acid (pH 5.4). To initiate crosslinking, 0.1% (w/v) sodium tripolyphosphate (STPP) solution gently added into chitosan solution with a constantly stirred using magnetic stirrer at room temperature. The colloidal suspension was centrifuged at 16,000 rpm for 30 minutes (40\u0026deg;C), and the resulting nanoparticles were lyophilized and washed with distilled water to ensure purity.\u003c/p\u003e\u003cp\u003eIn parallel, zinc oxide nanoparticles were prepared through pyrolysis. 50 g of zinc acetate dehydrate [Zn(CH\u003csub\u003e3\u003c/sub\u003eCOO)\u003csub\u003e2\u003c/sub\u003e\u0026middot;2H\u003csub\u003e2\u003c/sub\u003eO] was added to 25 g of tocopherol acetate together and heated at 70\u0026deg;C until it form homogenous solution. Then, the solution was heated to 120\u0026deg;C to evaporate residual solvents, yielding a viscous gel. Pyrolysis of the gel at 400\u0026deg;C for 5 hours in a muffle furnace produced crystalline ZnO nanoparticles.\u003c/p\u003e\u003cp\u003eChitosan nanoparticles (CHNPs 20 g) were dissolved in 100 mL of distilled water and 50 mL of 0.2 M acetic acid. The mixture was mixed with 50 mL of polyvinyl alcohol (PVA) and stirred for 5 minutes to form a homogeneous CH/PVA solution. To stabilize the composite, 4% glutaraldehyde was added as a crosslinking agent, followed by dropwise addition of potassium hydroxide solution (0.01 g in 10 mL H₂O) to form a homogenized system.\u003c/p\u003e\u003cp\u003e3.5% (w/v) zinc oxide nanoparticles (ZnO NPs) were added into the homogenized solution in a flask and kept in an oven for two days at room temperature. The samples obtained were washed with distilled water to remove undissolved glutaraldehyde at 25\u003csup\u003e0\u003c/sup\u003eC. Folic acid solution was obtained by dissolving 50ml of folic acid in 1M NaOH and was added to the homogenized solution by stirring at 45\u003csup\u003e0\u003c/sup\u003eC for 4 hours. Then, the final product CH/PVA/ZnO- FA was collected at room temperature.\u003c/p\u003e\u003c/div\u003e"},{"header":"3.0 Material Characterization","content":"\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003e3.1 X-ray Diffraction (XRD)\u003c/h2\u003e\u003cp\u003eX-ray diffraction (XRD) : This technique is used to analysis the crystalline nature of the synthesized material. The wavelength of the incident x-ray is determined by Bragg\u0026rsquo;s law in Eq.\u0026nbsp;1\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\:2dsin=n\\lambda\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\left(1\\right)$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003ewhere,\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\:d\\)\u003c/span\u003e\u003c/span\u003e denotes the distance between atomic planes (interplanar spacing), \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\theta\\:\\)\u003c/span\u003e\u003c/span\u003e is the angle of diffraction, \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\lambda\\:\\:\\)\u003c/span\u003e\u003c/span\u003erepresents wavelength, and \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:n\\)\u003c/span\u003e\u003c/span\u003e is an integer .\u003c/p\u003e\u003cp\u003eThe average size of crystalline index (\u003cem\u003eL\u003c/em\u003e) was estimated using the Scherrer formula in agreement with (Gumukaya \u003cem\u003eet al\u003c/em\u003e 2003, Popescus \u003cem\u003eet al\u003c/em\u003e 2011).\u003cdiv id=\"Equb\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equb\" name=\"EquationSource\"\u003e\n$$\\:L\\:=\\:\\frac{K\\:X\\:\\lambda\\:}{\\beta\\:\\:X\\:Sin\\:\\theta\\:}\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\left(2\\right)\\:$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003ewhere, \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:K\\)\u003c/span\u003e\u003c/span\u003e is a dimensionless shape factor (typically 0.91),\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\:\\lambda\\:\\)\u003c/span\u003e\u003c/span\u003e is the wavelength of the incident X-rays (0.154 nm),\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\:\\beta\\:\\)\u003c/span\u003e\u003c/span\u003eis the peak\u0026rsquo;s full width at half maximum (FWHM), and\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\:\\theta\\:\\)\u003c/span\u003e\u003c/span\u003e is the Bragg angle. This calculation provides a means of assessing the nanocrystalline features of the sample.\u003c/p\u003e\u003cp\u003eTo quantify the degree of crystallinity in the material, the crystallinity sizes was determined using this expression.\u003cdiv id=\"Equc\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equc\" name=\"EquationSource\"\u003e\n$$\\:{C}_{r}I=\\frac{{I}_{200}-{I}_{am}}{{I}_{200}}\\times\\:100\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\left(3\\right)$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003ewhere \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{I}_{200}\\)\u003c/span\u003e\u003c/span\u003e is the maximum intensity of the lattice diffraction and \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{I}_{am}\\)\u003c/span\u003e\u003c/span\u003e is the low intensity peak of amorphous region of the baseline at \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:2\\theta\\:\\)\u003c/span\u003e\u003c/span\u003e.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003e3.2 Scanning Electron Microscopy (SEM)\u003c/h2\u003e\u003cp\u003eA scanning electron microscope (SEM) determined the surface morphology of polymeric material. The image of the sample's surface at different areas or dimensions was determined using a scanning electron microscope (SEM).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003e3.3 Fourier Transform Infrared Spectroscopy (FTIR)\u003c/h2\u003e\u003cp\u003eFourier transform infrared (FTIR) spectroscopy is a technique used to obtain the functional groups present in the composite material. An infrared spectrum of absorption or emission was performed with a range of 400 to 4000 cm⁻\u0026sup1;, which helps to confirm the presence of characteristic chemical bonds and interactions among the components.\u003c/p\u003e\u003c/div\u003e"},{"header":"4.0 Results and Discussions","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\u003ch2\u003e4.1 Scanning Electron Microscopy of Polymeric Materials\u003c/h2\u003e\u003cp\u003eThe SEM micrographs in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec provide insight into the surface morphology and dispersion of zinc oxide nanoparticles (ZnO NPs) within various polymeric matrices, specifically chitosan/polyvinyl alcohol (CH/PVA), (CH/PVA/ZnO), and (CH/PVA/ZnO-FA) respectively. The image morphology of CH/PVA in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea reveals a spherical shape with a smooth loosely packed structures. This indicates an inhomogeneous surface typical of polymeric blends without nanofiller incorporation, this research is in supports of Cardo \u003cem\u003eet al\u003c/em\u003e., (2022). This structure may have impact in both the mechanical stability and drugs carrier if used in biomedical applications.\u003c/p\u003e\u003cp\u003eThe image surface CH/PVA/ZnO nanocomposite in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb, illustrates that the ZnO nanoparticles are dispersed throughout the polymer matrix, and enhancing surface uniformity. The particles appear more densely packed with increased surface roughness and micro-aggregates, which signifies successful incorporation of ZnO into the CH/PVA matrix. This improved morphology typically correlates with enhanced mechanical integrity, better thermal stability, and potentially higher drug-loading efficiency in agreement with Younes et al., (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eFigure \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec reveals further structural refinement. The surface becomes even more compact, with fewer visible and well-dispersed nanoparticles forming a consistent, densely packed layer. The folic acid conjugation likely promotes additional interactions within the matrix, reducing agglomeration and enhancing dispersion of ZnO nanoparticles. This modification is especially beneficial in targeted drug delivery, as folic acid improves cellular uptake by targeting folate receptors commonly over expressed in cancer cells. This result is in agreement with Tememel \u003cem\u003eet al.\u003c/em\u003e, (2012)\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003e4.2 Energy dispersive X-ray spectroscopy of Polymeric Materials\u003c/h2\u003e\u003cp\u003eThe Energy-Dispersive X-ray (EDX) spectroscopy analysis reveals discrete elemental compositions across the three polymer-nanoparticle formulations designed for targeted cancer drug delivery. The first spectrum in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea represents CH/PVA, which shows a predominant calcium content (45.0%), along with significant amounts of oxygen (25.0%), magnesium (15.3%), carbon (9.2%), phosphorus (3.5%), and sodium (2.0%). This baseline polymer blend exhibits no zinc content, providing a clear compositional reference point for the subsequent functionalized formulations, this report is in line with Agil-Abraham \u003cem\u003eet al.\u003c/em\u003e, (2016). The second spectrum in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb depicts CH/PVA/ZnO, which demonstrates the successful incorporation of zinc oxide nanoparticles, with zinc appearing prominently (30.0%) while maintaining calcium as a major component (38.0%). The relative proportions of other elements are adjusted accordingly with oxygen (15.0%), carbon (6.2%), magnesium (5.3%), phosphorus (3.5%), and sodium (2.0%). The introduction of zinc oxide nanoparticles significantly alters the elemental profile compared to the plain polymer blend, confirming effective nanoparticle integration within the polymeric matrix. Similar research by Tememel \u003cem\u003eet al\u003c/em\u003e., (2021) also revealed these properties. The third in spectrum in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ec corresponds to CH/PVA/ZnO-FA, which displays the highest zinc concentration (48.0%), indicating further enhanced zinc oxide nanoparticle loading after folic acid conjugation. This formulation shows a notable shift in calcium content (reduced to 15.3%) while maintaining substantial oxygen (20.0%) and carbon (10.2%) with lesser amounts of magnesium (4.5%) and sodium (2.0%), this result is in line with Tiyaboonchai, (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). The elevated zinc concentration in this final formulation suggests that the addition of folic acid may facilitate greater nanoparticle encapsulation or surface attachment, potentially enhancing the system's cancer-targeting capabilities through folate receptor-mediated mechanisms. This finding is consistent with Shanmuga \u003cem\u003eet al\u003c/em\u003e., (2013). The progressive increase in zinc content from CH/PVA to CH/PVA/ZnO-FA corresponds with the intended design of a drug delivery system with incrementally enhanced therapeutic potential for targeting cancer cells is in supports of Umesh \u003cem\u003eet al\u003c/em\u003e.,( 2011) article.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e(c) CH/PVA/ZnO \u0026ndash;FA\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003e4.3 X-ray Diffraction Analysis of Polymeric Materials\u003c/h2\u003e\u003cp\u003eFigure \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea displays characteristic crystalline peaks at 10.0\u0026deg;, 27.0\u0026deg;, 28.0\u0026deg;, 46.0\u0026deg;, 50.0\u0026deg;, and 69.0\u0026deg;, consistent with the crystalline phases of chitin and chitosan derived from crab shells, as well as the semi-crystalline nature of polyvinyl alcohol (PVA). These peaks correspond to the crystallographic planes (002), (100), (101), (200), (220), and (310), respectively, confirming the crystalline structure of the CH/PVA blend. The most intense peak at 27.0\u0026deg; (101) indicates a semi-crystalline polymer matrix, attributed to hydrogen bond between the two samples molecules. This report is in accord with Hang et al., (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2010\u003c/span\u003e); Nadia \u003cem\u003eet al\u003c/em\u003e., (2018).\u003c/p\u003e\u003cp\u003eThe XRD pattern of CH/PVA/ZnO exhibits distinct peaks at 10.0\u0026deg;, 27.5\u0026deg;, 28.0\u0026deg;, 30.0\u0026deg;, 40.0\u0026deg;, 60.0\u0026deg;, and 65.5\u0026deg;in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb, assigned to the lattice planes (002), (100), (101), (200), (220), (300), and (311), respectively. The prominent peak at 28.0\u0026deg; (101) confirms the crystalline structure of ZnO nanoparticles embedded within the polymeric matrix. The presence of these peaks suggests structural modification due to the interaction between chitosan, PVA, and ZnO, leading to the formation of new crystalline phases or polymorphs. Similar research conducted by Casettari \u003cem\u003eet al\u003c/em\u003e., (2013) ; Gocho\u003cem\u003eet al\u003c/em\u003e., (2000); Kardas \u003cem\u003eet al\u003c/em\u003e., (2012) revealed same properties. The XRD pattern of CH/PVA/ZnO-FA (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ec) reveals peaks at 10.0\u0026deg;, 26.0\u0026deg;, 28.5\u0026deg;, 30.0\u0026deg;, 40.0\u0026deg;, 50.0\u0026deg;, and 70.0\u0026deg;, corresponding to the planes (100), (002), (101), (200), (220), (300), and (311). The most intense peak at 28.5\u0026deg; (002) signifies enhanced crystallinity and uniform atomic stacking along the c-axis. This preferred orientation indicates improved crystalline alignment, which influence material properties such as mechanical strength and thermal stability. However, variations in peak intensity may also introduce micro strain or lattice defects due to atomic vacancies, potentially reducing crystallite size. This result is in agreement with (Nagarwal \u003cem\u003eet al\u003c/em\u003e., 2012; Zaman \u003cem\u003eet al\u003c/em\u003e., 202; D-campose \u003cem\u003eet al\u003c/em\u003e., 2004; Xie \u003cem\u003eet al\u003c/em\u003e., 2007; Herdiana \u003cem\u003eet al\u003c/em\u003e., 2021).\u003c/p\u003e\u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e4.1\u003c/span\u003e indicates that the crystallinity index of CH/PVA/ZnO-FA is 78%, which is higher than CH/PVA and CH/PVA/ZnO. Other parameters, including crystallite size (L), d-spacing (d) and full width at half maximum (FWHM). The increased crystallinity index of CH/PVA/ZnO-FA significantly enhances its physical, mechanical, and thermal properties of the samples and offers potential for a more sustained release profile, making it a promising polymer for carcinogenic applications.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 4.1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eCrystallographic Parameters Obtained from XRD Patterns\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eS/N\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSample\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e2θ (\u003csup\u003e0\u003c/sup\u003e)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003ed (nm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eL (nm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eFWHM\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{C}_{r\\:}\\)\u003c/span\u003e\u003c/span\u003eI (%)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1\u003c/p\u003e\u003cp\u003e2\u003c/p\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCH/PVA\u003c/p\u003e\u003cp\u003eCH/PVA/ZnO\u003c/p\u003e\u003cp\u003eCH/PVA/ZnO\u0026ndash;FA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e27.0\u003c/p\u003e\u003cp\u003e28.0\u003c/p\u003e\u003cp\u003e28.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.66\u003c/p\u003e\u003cp\u003e1.71\u003c/p\u003e\u003cp\u003e2.65\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.35\u003c/p\u003e\u003cp\u003e0.28\u003c/p\u003e\u003cp\u003e0.27\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.50\u003c/p\u003e\u003cp\u003e0.51\u003c/p\u003e\u003cp\u003e0.52\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e72\u003c/p\u003e\u003cp\u003e74\u003c/p\u003e\u003cp\u003e78\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003e4.4 Fourier Transform (FTIR) Spectral Analysis of Polymeric Materials\u003c/h2\u003e\u003cp\u003eFTIR spectroscopic analysis of the synthesized polymeric materials in Figs.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, show their molecular architecture and intermolecular interactions that illustrate the drug delivery functionality.\u003c/p\u003e\u003cp\u003eThe absorption band of the CH/PVA polymer blend is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The broad absorption band at 3438 cm⁻\u0026sup1; corresponds to the stretching vibrations of hydroxyl (O-H) and amine (N-H) groups respectively. This peak's relatively sharp profile suggests increased polymer chain interactions may enhance drug carrier capacity. The peak values observed at 2926 cm⁻\u0026sup1; and 2400 cm⁻\u0026sup1; confirm the presence of alkyl (C-H) stretching, whereas those between 1775\u0026ndash;1131 cm⁻\u0026sup1; indicate the presence of carbonyl (C\u0026thinsp;=\u0026thinsp;O) groups from acetate. The observed minor peak shifts compared to bulk materials indicate modified molecular interactions resulting from nanoscale particle formation, which could optimize drug loading efficiency, This result seen is agreement with similar studies by Ndamitso \u003cem\u003eet al.,(\u003c/em\u003e2020); Zhong et al., (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe CH/PVA/ZnO composite exhibits intensified absorption features. The broad absorption band of O-H/N-H stretching vibrations at 3440 cm⁻\u0026sup1; shows increased intensity and a transmittance level of 38%, suggesting enhanced hydrogen bonding. The amide I band shift to 1633 cm⁻\u0026sup1; reflects structural modifications from ZnO incorporation. The maintained peaks at 2926 cm⁻\u0026sup1; (C-H) and 1039 cm⁻\u0026sup1; (C-O) verify retained glutaraldehyde crosslinks. These spectral changes indicate a strong intermolecular interactions that may improve drug encapsulation stability, this aligns with findings from Sumaila \u003cem\u003eet al.,(\u003c/em\u003e 2020); Mohammed \u003cem\u003eet al\u003c/em\u003e., (2019).\u003c/p\u003e\u003cp\u003eFigure \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e reveals the FTIR profile of the FA-conjugated system. The broad absorption band of 3442cm⁻\u0026sup1; confirms the presence of persistent strong hydrogen bond which is crucial for drug-polymer interactions. Key features include the amide I band, C-H, and C-O band reveal modified glycosidic linkages. These spectral modifications confirm successful F-A functionalization while maintaining the polymer's structural integrity, essential for targeted drug delivery, similar research by Mohammad-pourd, \u003cem\u003eet al.,(\u003c/em\u003e2010);Tolaimate et al., (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2003\u003c/span\u003e) revealed same result.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003ch2\u003e4.5 Thermogravimetric analysis of Polymeric Material\u003c/h2\u003e\u003cp\u003eThe TGA curves for CH/PVA, CH/PVA/ZnO, and CH/PVA/ZnO-FA reveal distinct thermal behaviors that reflect the influence of ZnO nanoparticles and folic acid (FA) on the polymer matrix. For the CH/PVA sample (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ea), the initial weight loss observed below 100\u0026deg;C is minimal and corresponds to the evaporation of adsorbed moisture, as indicated by weight retention of 99.7% at 96\u0026deg;C. The main decomposition event occurs between approximately 300\u0026deg;C and 400\u0026deg;C, where the polymer undergoes rapid weight loss. The sharp decline in the temperature observed signifies the breakdown of the chitosan/PVA backbone, with a major decomposition temperature around 375\u0026deg;C, where 70.5% of the mass is lost. This report is in agreement with Agnihotri et al., (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). In addition, incorporating ZnO nanoparticles into the CH/PVA matrix, as shown in the CH/PVA/ZnO sample (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eb), slightly enhances the thermal stability of the composite, the onset of decomposition remains in the same temperature range, but the weight loss 72.2% occurs at 382\u0026deg;C, indicating a marginal increase in thermal resistance. The residue at 450\u0026deg;C remains at 10.75%, suggesting that ZnO nanoparticles, being inorganic, contribute to the non-volatile residue. The improved thermal stability can be attributed to the strong interfacial interactions between the polymer chains and ZnO nanoparticles, which restrict the mobility of the polymer chains and delay thermal degradation. This enhancement is particularly beneficial for drug delivery applications, as it ensures that the nanocomposite carrier can withstand higher processing and physiological temperatures without premature degradation. This result is similar to Somnuk et al., (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2011\u003c/span\u003e); Tian \u003cem\u003eet al.,(\u003c/em\u003e 2014) reports\u003c/p\u003e\u003cp\u003eThe CH/PVA/ZnO-FA sample (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ec) reveals a slightly different thermal profile. The major decomposition temperature observed at 350\u0026deg;C and 70.5% weight loss are lower than the CH/PVA/ZnO sample. This slight reduction in thermal stability may be due to the introduction of folic acid, which, enhancing the targeting capability of the nanocomposite for cancer cells. Moreover, the residue at 450\u0026deg;C remains consistent at 10.75%, indicating that the inorganic content (ZnO) still dominates the final residue. The presence of folic acid introduced has a significant functional advantage for drug delivery system, even if it slightly compromises thermal stability, this finding is in agreement with Gupta and Kompella (2010); Cheba, (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2011\u003c/span\u003e); Ali et al., (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) reports.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe comprehensive characterization of CH/PVA, CH/PVA/ZnO, and CH/PVA/ZnO-FA nanocomposites reveals a systematic enhancement of structural, elemental, crystallographic, molecular, and thermal properties tailored for anticancer drug delivery. SEM analysis illustrates progressive morphological refinement from the porous CH/PVA matrix to the optimally dispersed FA-conjugated system, where improved nanoparticle distribution and reduced defects enhance drug-loading capacity and release kinetics. Complementary EDX results confirm the evolution through increasing zinc content (0% \u0026rarr; 30% \u0026rarr; 48%), validating successful ZnO incorporation and FA-mediated nanoparticle enrichment while maintaining polymer backbone integrity through consistent oxygen/carbon signatures. XRD patterns corroborate these findings, showing enhanced crystallinity (peaking at 43.3% for CH/PVA/ZnO-FA) and atomic ordering through distinct diffraction planes, particularly the intensified 28.5\u0026deg; (002) peak indicating structural perfection. FTIR spectroscopy reveals preserved hydrogen bonding networks and new vibrational modes, confirming molecular stability while demonstrating strengthened polymer-nanoparticle interactions and successful FA functionalization. Thermal analysis completes this picture, showing ZnO's stabilizing effect (382\u0026deg;C decomposition temperature) and FA's strategic trade-off between modest thermal reduction (350\u0026deg;C) and gained targeting capability. Together, these characterizations prove the nanocomposites achieve an optimal balance: morphological uniformity for drug loading, elemental specificity for therapeutic action, crystallographic stability for mechanical strength, molecular precision for controlled release, and thermal resilience for processing - all while incorporating cancer-targeting functionality. The FA-conjugated system emerges as particularly promising, combining structural advantages with biological targeting through folate receptors. This multidisciplinary validation confirms the nanocomposites' potential as advanced, multifunctional platforms for targeted carcinogenic disease treatment, where material properties align precisely with therapeutic requirements\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003e\u0026nbsp;2 Funding Declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research did not receive any specific funding or grant from institution \u0026ndash; based or non-governmental organization.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3 Clinical Trial Number\u003c/strong\u003e: Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e4 Consent to Publish Declaration\u003c/strong\u003e: Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e5 Ethics and Consent to Participate Declarations\u003c/strong\u003e: Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e6 Authors Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eL.O.A: Conceptualization, investigation, Visualization, Writing \u0026ndash; Original Draft Preparation\u003c/p\u003e\n\u003cp\u003eA.F.A: Formal Analysis, Writing-Reviewing and Editing.\u003c/p\u003e\n\u003cp\u003eM.A.A: Methodology, Draft Reviewing, Editing and Discussion\u003c/p\u003e\n\u003cp\u003eO.I.O: Methodology, Formal Analysis, Draft Reviewing, and Supervision.\u003c/p\u003e\n\u003cp\u003eS.S.O : Project Administration, Methodology, Validation and Supervision.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e7 Competing interest Declaration\u003c/strong\u003e: No completing interest exists in this work.\u003c/p\u003e\n\u003cp\u003e8 \u003cstrong\u003eAuthor Email Discrepancy\u003c/strong\u003e: \u0026nbsp;[email protected]\u003c/p\u003e\n\u003cp\u003e9 \u003cstrong\u003eCity in Affiliation\u003c/strong\u003e: \u003csup\u003eb\u003c/sup\u003eDepartment of Physics, Wigwe University, Isiokpo, Rivers State, Nigeria.\u003c/p\u003e\n\u003cp\u003e10 \u003cstrong\u003eAcknowledgements:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eI sincerely wish to express my profound gratitude to the department of Physics, Federal University of Technology, Akure, for their valuable contribution during the material preparation. My special thanks go to Dr. Francis Alo of the Department of Material science and Engineering, Obafemi Awolowo University, Ile \u0026ndash;Ife, Nigeria, for his immense support during the material characterization.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDecision: Minor revision\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e1 \u003cstrong\u003eData Availability Statement\u003c/strong\u003e:\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp;All data generated or analysed \u0026nbsp;during this study are included in this published article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2 Permissions to collect the crab shells:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCrab shells are available in area surrounding our local river, therefore, we don\u0026rsquo;t need licenses for \u0026nbsp; \u0026nbsp;collection of shells.\u003c/p\u003e\n\u003cp\u003e3 \u003cstrong\u003eArticle Type:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp;Thanks for the observation. This a pure research works.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDecision: Minor revision on 13 oct 2025\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e1 Revise title:\u0026nbsp;\u003c/strong\u003eSynthesis, Characterization and Potential Drug Delivery Applications of Polymer-Coated Zinc Oxide Nanoparticles for Cancer-Targeted Therapy\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2 Data availability statement\u003c/strong\u003e:\u003cbr\u003e\u0026nbsp;\u003cbr\u003e\u0026nbsp;The submission system under the \u0026apos;Declaration\u0026apos; section by following \u0026nbsp;step: Done\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbhinay S, Kumar D, Meenakshi A Dua., Gursharan, Singh, Chhatwal and AtulKuma, Johr. (2012). 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Journey Pharm Sci 32:171\u0026ndash;175\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAliasghari A, Khorasgani MR, Vaezifar S, Rahimi F, Younesi H (2016) Evaluation of antibacterial efficiency of chitosan and chitosan nanoparticles on cariogenic streptococci: An in vitro study. \u003cem\u003eIran. Journal Microbiology., 8, 93\u0026ndash;100.\u003c/em\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCheba BA (2011) Chitin and Chitosan: Marine Biopolymers with Unique Properties and Versatile Applications. Global J Biotechnol Biochemisry 6:149\u0026ndash;153\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDutta PK, Dutta J, Tripathi VS (2004) Chitin and chitosan: chemistry, properties and applications. Journal of Scientific and Industrial Research page 63, 20\u0026ndash;31.edible mushrooms,Nile perch scales and banana weevils for biomedical applications. \u003cem\u003eJournal of Scientific, Report. 1, 4116.\u003c/em\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGupta RB, Kompella UB (2006) Fundamentals of drug Nanoparticles Technology for Drug Delivery. \u003cem\u003eNew York: Taylor \u0026amp; Francis group; p pp. 1\u0026ndash;18.\u003c/em\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHang AT, Tae B, Park JS (2010) Non-woven mats of poly(vinyllcohol)/chitosan blends containing silver nanopartiocles: Fabrication and haracterization. \u003cem\u003eCarbohydrate Polymer. 82:472479\u003c/em\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMohammad pourdounighi N, Behfar A, Ezabadi A, Zolfagharian H, Heydari M (2010) Preparation of Chitosan nanoparticles containing \u003cem\u003eNaja-najaoxiana\u003c/em\u003esnake venom.\u003cem\u003eJournal of Nanomedicine.;6(1):137\u0026thinsp;\u0026ndash;\u0026thinsp;43.\u003c/em\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMohammed M, Mansell H, Shoker A, Wasan KM, Wasan EK .(2019)Development and in vitro characterization of chitosan-coated polymeric nanoparticles for oral delivery and sustained release of release of the immunosuppressant drug mycophenolate mofetil. Drug Dev Ind Pharm ;\u003cem\u003e45\u003c/em\u003e(\u003cem\u003e1\u003c/em\u003e):76\u0026ndash;87\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eTian X, Yin H, Zhang S (2014) Bufalin loaded Biotinylated Chitosan Nanoparticles as an efficient drug delivery system for targeted chemotherapy against breast carcinoma. Eur J Pharm ( 87:445\u0026ndash;453\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eTiyaboonchai W (2003) Chitosan nanoparticles: a promising system for drug delivery. NaresuanUniversity J 11(3):51\u0026ndash;66\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eTolaimate A, Desbrieres J, Rhazi MAA (2003) C haracterization and preparation of chitins and chitosans with controlled physico-chemical properties. J Polym Sci 44:79397952 ]24\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSomnuk J, Anupap T, Virote B (2011) Preparation of chitosan nanoparticles for encapsulation and release of protein. Korean journal Chem Engineering 28(5):1247\u0026ndash;1251\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eXing K, Chen XG, Kong M, Liu CS, Cha DS, Park HJ (2009b) Effect of oleoyl chitosan nanoparticles as a novel antibacterial dispersion system on viaibilitymembrane permeability and cell morphology of Escherichia coli and Staph lococcus. \u003cem\u003eJournal of Carbohydrate Polymers page76, 17\u0026ndash;22.\u003c/em\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eYounes I, Ghorbel-Bellaaj O, Nasri R, Chaabouni M, Rinaudo M, Nasri M (2012) Chitin and chitosan preparation from shrimp shells using optimized enzymatic deproteinization. \u003cem\u003eB Journal on Process Biochemistry ;47:2032\u0026ndash;2039.\u003c/em\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eYounes I, Hajji S, Frachet V, Rinaudo M, Jellouli K, Nasri M (2014) Chitin extraction from shrimp shell using enzymatic treatment. Antitumor, antioxidant and antimicrobial activities of chitosan. \u003cem\u003eInternational Journal Biology Macromolecule 69, 489\u0026ndash;498.\u003c/em\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eYounes I, Rinaudo M (2015) Chitin and chitosan preparation from marine sources, Structure properties and applications. 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Nanopart Polym 10(4):462\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhao LM, Shi LE, Zhang ZL, Chen JM, Shi DD, Yang J, Tang Z X.(201 Preparation and application of chitosan nanoparticles and nanofibers; Brazilian \u003cem\u003eJournal of Chemical Engineering, 28,. (03). 353\u0026ndash;362.\u003c/em\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhong Z, Xing R, Liu S, Wang L, Cai S, Li P (2008) Synthesis of acyl thiourea derivatives of chitosan and their antimicrobial activities in vitro. \u003cem\u003eJournal of Carbohydrate Resource 343:566\u003c/em\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhong Z, Chen R, Xing R, Chen X, Liu S, Guo Z, Ji X (2018) Synthesis and antifungal properties of sulfanilamide derivatives of chitosan. Elsevier Ltd. All rights reserved\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCardoso G, Sirlene MM, Pereira W, Lopes MA (2022) \u003cem\u003eComposition Effects on the MorphologyofPVA/Chitosan Electrospun Nanofibers\u003c/em\u003e. \u003cem\u003e14\u003c/em\u003e(22), 4856\u0026ndash;4856. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003edoi.org/10.3390/polym14224\u003c/span\u003e\u003cspan address=\"10.3390/polym14224\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"discover-nano","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"narl","sideBox":"Learn more about [Discover Nano](https://www.springer.com/journal/11671)","snPcode":"11671","submissionUrl":"https://submission.nature.com/new-submission/11671/3","title":"Discover Nano","twitterHandle":"@SpringerOpen","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Crab shells, Ionic gelation, Chitosan, Nanoparticles, Zinc oxide, Polymer, and Folic acid","lastPublishedDoi":"10.21203/rs.3.rs-7602894/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7602894/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study developed a chitosan-polyvinyl alcohol nanocomposite system (CH/PVA) incorporated with zinc oxide nanoparticles (ZnO) and conjugated with folic acid (FA) for targeted drug delivery applications. The synthesized materials CH/PVA, CH/PVA/ZnO, and CH/PVA/ZnO-FA were thoroughly characterized using multiple analytical techniques. Scanning electron microscopy (SEM) revealed that the folic acid-conjugated sample (CH/PVA/ZnO-FA) exhibited a distinct bead-like morphology, suggesting enhanced drug-loading capacity and therapeutic potential. Energy-dispersive X-ray spectroscopy (EDX) confirmed the elemental compositions (Zn, O, Mg, Na, C and Ca), showing prominent zinc signals (48.0 wt %) in CH/PVA/ZnO-FA, indicating successful nanoparticle incorporation and FA conjugation. X-ray diffraction (XRD) analysis demonstrated high crystallinity, with a strong diffraction peak at 28.5\u0026deg; corresponding to the (002) plane, reflecting optimal atomic packing density. Fourier-transform infrared spectroscopy (FTIR) shows functional groups of the samples including hydroxyl (-OH) and amine (-NH) stretching vibrations at 3440 cm⁻\u0026sup1;, confirming increase in hydrogen bond within the polymer matrix. Thermogravimetric analysis (TGA) revealed excellent thermal stability up to 350\u0026deg;C, suitable for biomedical applications. These comprehensive characterization results demonstrate that the CH/PVA/ZnO-FA nanocomposite possesses ideal structural, chemical, and thermal properties for use as an advanced nanocarrier system in targeted cancer therapy applications.\u003c/p\u003e","manuscriptTitle":"Synthesis, Characterization and Potential Drug Delivery Applications of Polymer-Coated Zinc Oxide Nanoparticles for Cancer-Targeted Therapy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-03 07:13:16","doi":"10.21203/rs.3.rs-7602894/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-03-18T20:03:51+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-07T11:55:57+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-04T04:57:12+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-03T13:29:36+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"267429600862079634531959237973818517948","date":"2025-12-29T10:35:31+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"158369286122330396489989433335462470923","date":"2025-12-28T23:52:44+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-27T17:08:39+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-27T13:52:36+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"339916622195325052645893378118404363646","date":"2025-12-26T06:32:42+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"38176806117597014095344067828127928969","date":"2025-12-25T20:13:53+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"54659663910502472363651757089409059165","date":"2025-12-25T14:23:08+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-01T09:20:00+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"189885114340372541485675087159307886706","date":"2025-11-22T07:16:10+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"212177996347421727936318710447987428448","date":"2025-11-21T14:29:39+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"286788881359023729804127703267233417561","date":"2025-11-21T11:47:04+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-19T17:17:28+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"129054934820466424925339651380396281317","date":"2025-11-10T07:00:18+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"94605254813258871332001106406089932169","date":"2025-11-10T05:34:06+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-10T04:16:50+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-10-14T11:34:53+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-10-11T15:21:02+00:00","index":"","fulltext":""},{"type":"submitted","content":"Discover Nano","date":"2025-10-11T15:16:47+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"discover-nano","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"narl","sideBox":"Learn more about [Discover Nano](https://www.springer.com/journal/11671)","snPcode":"11671","submissionUrl":"https://submission.nature.com/new-submission/11671/3","title":"Discover Nano","twitterHandle":"@SpringerOpen","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"c9e4f5f7-1b72-45a0-be99-ff7ef8761aee","owner":[],"postedDate":"November 3rd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-05-12T06:08:19+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-03 07:13:16","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7602894","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7602894","identity":"rs-7602894","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}