STRUCTURAL ELUCIDATION AND NANOPARTICLE FORMULATION OF TYROSOL FOR BIOMEDICAL APPLICATIONS

Abstract

Background Tyrosol, 2-(4-hydroxyphenyl)ethanol) is a pharmacologically active phenolic compound. Nanobiotechnology-based drug delivery system offers the advantage of small size, enhanced cellular uptake and internalization for site-specific delivery. Several drug functionalized nanocarrier delivery systems have been formulated to limit cellular drug toxicity, enhanced therapeutic efficacy and improved pharmacokinetics. Nowadays, biogenic nanoparticles unveil distinctive physicochemical properties appropriate for their application in disease diagnosis and treatment. Aims In this study, we isolated and structurally characterized the tyrosol from olive oil and performed its conformational analysis and preferential COX-2 inhibition using in silico methods. Tyrosol is a hydrophilic molecule that undergoes rapid first-pass metabolism leading to low systemic bioavailability. We have developed tyrosol functionalized chitosan gold nanoparticles (Chi-TY-AuNP's) to overcome first-pass intestinal susceptibility and aid in sustained drug release. Further, we evaluated its antidiabetic, antibiofilm and wound healing efficacy. Materials and Methods Conformational analysis of isolated tyrosol was carried out using density functional theory. In silico ADMET, molecular docking and molecular dynamics simulations of tyrosol with COX-2 receptor were performed. Further, Chi-TY-AuNP’s were synthesized and characterized using state-of-the-art biophysical techniques. We examined the in vitro cytotoxicity and ROS generation using RIN-5F cell lines and in vivo antidiabetic activity of Chi-TY-AuNP's by streptozotocin-induced diabetes in Wistar albino rats. Changes in bodyweight, hematological and lipid parameters along with histopathological features were analyzed. The antibiofilm potency of Chi-TY-AuNP's was investigated by examining their impact on ROS generation, cell surface hydrophobicity, ECM composition and membrane ergosterol content in biofilms of C. albicans and C. glabrata, besides, transcriptional expression of selected C. glabrata genes was also evaluated. Furthermore, in vitro cytotoxicity and ROS generation analysis of Chi-TY-AuNP's using NIH-3T3 cell lines and in vivo wound healing activity of Chi-TY-AuNP’s-Carbopol®934 gel was evaluated using the excision wound model in Wistar albino rats. The healing efficacy was measured in terms of percentage wound contraction, epithelialization period and histopathological studies. Results In silico analysis showed that tyrosol possesses lead/ drug-like potential for COX-2. Chi- TY-AuNP’s synthesis was ascertained by the appearance of ruby red color and consistent surface plasmon band peaks at 535 nm. Biophysical characterization confirms the high colloidal stability of spherical Chi-TY-AuNP’s. Chi-TY-AuNP’s showed a biphasic drug release profile, an initial burst in the first 6 h followed by sustained release of tyrosol. In vitro studies revealed good cytocompatibility and reduced ROS levels in RIN-5F cells treated with Chi-TY-AuNP’s. Chi-TY-AuNP’s treated diabetic rats showed the ameliorating effect on blood glucose level and biochemical parameters in a dose-dependent manner. Histopathology of treated rat’s pancreas manifested restored islets of Langerhans and pancreatic acini. Chi-TY-AuNP’s showed fungicidal and biofilm eradication potential against both the Candida spp. Biochemical studies revealed the interference of Chi-TYAuNP’s with generated ROS, ECM components and ergosterol content in Candida biofilms along with downregulation of selected C. glabrata genes involved in the maintenance of cell wall biosynthesis. Chi-TY-AuNP’s treatment revealed good cytocompatibility and decreased ROS levels in NIH-3T3 cells. In vivo wound healing study demonstrated the reepithelialization of the wounded tissues and increased proliferation of fibroblast cells, collagen fibers and neoangiogenesis. Conclusion Chi-TY-AuNP’s manifested greater colloidal stability and high positive surface charge. Conclusively the result suggested good cytocompatibility of Chi-TY-AuNP’s and significant antidiabetic, antibiofilm and wound healing efficacy. Some challenges remain to be addressed, such as long-term stability and toxicity analysis, pharmacokinetics and biodistribution studies of Chi-TY-AuNP’s. Future Prospects The finding of this study suggested Chi-TY-AuNP’s efficiency for the management of diabetes, cutaneous wounds and Candida related infections. Further research is warranted for the clinical translation of Chi-TY-AuNP’s as an alternative strategy for its safe and diverse clinical applications.