“IN VITRO WOUND HEALING POTENTIAL OF AZADIRACHTA INDICA–LIGNOCAINE GEL: ENHANCING FIBROBLAST MIGRATION AND REGENERATION FOR WOUND MANAGEMENT”

Abstract

Background: Wound healing is a complex process involving haemostasis, inflammation, proliferation, and remodelling. Herbal formulations have gained interest due to their antioxidant and regenerative properties. Azadirachta indica (neem) exhibits antimicrobial, antioxidant, and tissue-regenerative effects, while lignocaine offers local analgesia and anti-inflammatory activity. This study aimed to develop a stable neem-lignocaine gel and evaluate its in vitro wound healing potential using fibroblast proliferation, collagen production, and scratch assay closure.

Methods: Neem extract was prepared by combining 5 g of Azadirachta indica powder with 100 mL distilled water, heating at 50 °C for 20 min, filtering, and concentrating to 10 mL. Mouse 3T3-L1 fibroblasts were cultured for cytotoxicity and wound healing studies. The MTT assay assessed cell viability at 25–150 μg/mL over 24 h. Scratch assays were performed on confluent fibroblast monolayers treated with 100 μg/mL gel. Wound closure was monitored and quantified using ImageJ software.

Results: MTT assay showed dose-dependent cytotoxicity. Cell viability remained above 95% at 25–75 μg/mL, over 80% at 100 μg/mL, and decreased to 70–75% at 125–150 μg/mL, confirming acceptable safety at therapeutic concentrations. Scratch assay demonstrated accelerated wound closure with neem-lignocaine gel, indicating enhanced fibroblast migration and proliferation compared to controls, which exhibited slower closure with persistent gaps.

Conclusion: Neem-lignocaine gel promotes fibroblast proliferation and wound closure in vitro, combining the regenerative potential of herbal medicine with the analgesic and anti-inflammatory benefits of lignocaine. This formulation represents a cost-effective and clinically promising strategy for wound management.