Green Innovation in GTR Membranes: Combining Astaxanthin, <i>Aloe vera</i> and Chitosan for Periodontal Therapy- An <i>In Vitro</i> Study

Background: Periodontitis leads to tissue deterioration, prompting the need for effective regenerative therapies. Conventional barrier membranes, face challenges in mechanical strength and support. This study explores a novel formulation combining Astaxanthin’s antioxidant and anti-inflammatory properties, Aloe vera’s healing effects, chitosan nanoparticles biocompatibility and antimicrobial benefits to enhance periodontal tissue regeneration. Aim: This study aimed to develop a Guided Tissue Regeneration (GTR) membrane incorporating Astaxanthin, Aloe vera, and chitosan nanoparticles and evaluate its in vitro properties. Methods: Astaxanthin and Aloe vera extracts were dissolved in deionized water and boiled for complete incorporation. Chitosan nanoparticles were created by dissolving chitosan in acetic acid, followed by the drop wise addition of sodium tripolyphosphate while stirring. The nanoparticles were collected via centrifugation, washed, and re-suspended in deionized water. A chitosan solution was then prepared with glycerol as a plasticizer, and a cross-linking agent was added for stabilization. Finally, the chitosan nanoparticles and bioactive extracts were blended to form the membrane and later the same was analyzed for Scanning Electron Microscopic (SEM), Fourier Transform Infrared Spectroscopy (FTIR), cytotoxic, anti-oxidant, and degradation test activities. Results: The GTR membrane demonstrated a homogeneous nanoparticle distribution and initial degradation signs in SEM analysis, while FTIR confirmed the integration of Astaxanthin, Aloe vera, and Chitosan. Cytotoxicity tests showed low toxicity, with significant MG-63 cell viability at 15, 20, and 25 μg/mL. Antioxidant activity was dose-dependent, peaking at nearly 50% scavenging at 25 μg/mL, though still lower than ascorbic acids 65%. The degradation test revealed an increasing rate with concentration, reaching approximately 40% at 25 μg/mL. Conclusion: Thus the formulated GTR membrane effectively integrates Astaxanthin, Aloe vera, and chitosan nanoparticles, demonstrating promising in vitro properties for periodontal tissue regeneration with low cytotoxicity and significant antioxidant activity suitable for regenerative therapies. Major Findings: The developed GTR membrane successfully incorporated with Astaxanthin, Aloe vera, and chitosan nanoparticles, exhibited favourable in vitro characteristics for periodontal tissue regeneration. It demonstrated minimal cytotoxicity and notable antioxidant activity, highlighting its potential for periodontal therapeutic applications.