Abstract
Pathogenic microorganisms, ovarian cancer, and inflammation represent significant health challenges, as infections can disrupt the vaginal microbiome, chronic inflammation contributes to cancer progression. This study explored the enhancement of the therapeutic efficacy of vaginal wash (VW) incorporate chitosan nanoparticles (Chit) (VW + Chit) to create a multifunctional formulation targeting microbial infections, inflammation, and ovarian cancer. Gas Chromatography-Mass Spectrometry analysis identified 42 chemical constituents in the vaginal wash, including bioactive alcohols, esters, ethers, and fatty acids, with major components including 1,2-benzenedicarboxylic acid, diethyl ester (24.83%) and Diethylene glycol monododecyl ether (7.10%). Transmission electron microscopy imaging revealed that VW + Chit nanoparticles exhibited improved morphology, reduced agglomeration, and increased particle size (~ 41 nm), indicating successful interaction and encapsulation. Fourier transform infrared spectroscopy confirmed strong intermolecular bonding between chitosan and vaginal wash components, suggesting enhanced structural stability. Antimicrobial testing showed that VW + Chit achieved significantly greater inhibition zones and lower MIC/MBC/MFC values across multiple pathogens, particularly Staphylococcus aureus, Enterococcus faecalis, and Candida albicans, outperforming both its individual components and standard antibiotics. The formulation VW + Chit also demonstrated potent anti-inflammatory activity, with COX-1 and COX-2 inhibition reaching 92.9% and 94.3%, with lowest IC₅₀ values 20.92 ± 0.1 and 9.23 ± 0.1 µg/mL, respectively compared to IC₅₀ values of utilize VW (47.52 ± 0.4 and 43.73 ± 0.1 µg/mL) and Chit alone (27.39 ± 0.1 and 21.67 ± 0.1 µg/mL), respectively. Cytotoxicity assays on SKOV3 ovarian cancer cells revealed that VW + Chit exerted the strongest dose-dependent cytotoxic effect, with the lowest IC₅₀ (147.3 µg/mL) and significant G1 phase cell cycle arrest, indicating antiproliferative potential. In conclusion, the VW + Chit composite shows enhanced antimicrobial, anti-inflammatory, and anticancer activities, highlighting its potential as a novel therapeutic formulation. These findings warrant further investigation through formulation refinement and in vivo studies to validate its clinical applicability for gynecological use.