BIOLOGICAL EVALUATION OF ALPHA-TOCOPHEROL-CHITOSAN BIO-COMPOSITE: ANTI-INFLAMMATORY, ANTIOXIDANT, AND TOXICOLOGICAL ASSESSMENTS USING ZEBRAFISH EMBRYOS

Objective: This study aimed to synthesize and evaluate a novel eco-friendly α-tocopherol–chitosan (CS) bio-composite for potential medicinal applications, emphasizing its thrombolytic, cytotoxic, anti-inflammatory, and antioxidant properties. Methods: The α-tocopherol-CS bio-composite was synthesized and initially confirmed by color changes and visual observation. CS and the resulting bio-composite were characterized by ultraviolet-visible (UV-Vis) spectroscopy, which showed distinct peaks at 362 and 350 nm. Anti-inflammatory activity was assessed using bovine serum albumin, egg albumin, and membrane stabilization assays at 10–50 μg/mL concentrations. Antioxidant activity was analyzed statistically using two-way Analysis of Variance. Cytotoxicity was evaluated by the brine shrimp lethality assay, while anticoagulant and thrombolytic properties were measured by clot lysis at 50 μg/mL. Results: UV-Vis spectroscopy validated the formation of the α-tocopherol-CS nanoformulation. The bio-composite demonstrated inhibition rates of 41–76% (bovine serum albumin), 50–74% (egg albumin), and the highest inhibition of 52–84% and an IC50 value of 5 μg/mL (membrane stabilization). Antioxidant activity was highly significant (time, p