Reverse pharmacology approach to validate the diabetic wound healing activity of Jatyadi thailam formulations an in vitro study

Abstract

Diabetes-related chronic wounds are a leading cause of disability and death newlineglobally, especially in India, due to a high prevalence of foot ulcers, often resulting in newlineamputations. This study employs a Reverse Pharmacology approach to analyze Jatyadi newlinethailam, a recognized Ayurvedic wound-healing and anti-inflammatory preparation. We newlineinvestigated two Jatyadi thailam formulations - JT-AFI and JT-YG, for their newlinepharmacological mechanisms. Both formulations are soluble in a mixture of PEG-40 HCO, newlineSPAN 80, and water. The study comprises four phases: Phase I involved assessment of newlinebiological activity, including free radical scavenging, biomolecule protection, antimicrobial, newlineand anti-inflammatory properties in vitro, Phase II investigated the dermal safety using newlinerats, examining skin irritation potential and sub-acute toxicity tests. Phase III evaluated the newlinediabetic wound-healing properties using L929 cell line, including cell survival, proliferation, newlinemigration, angiogenesis, cell cycle regulation, apoptosis, ROS generation, and newlinemitochondrial membrane potential. In Phase IV, we selected the superior formulation and newlineemployed a network pharmacology approach to identify its components and targets newlinerelated to diabetic wound healing, which was subsequently validated through Western blot newlineanalysis. Results show that both formulations effectively scavenged free radicals and newlineprotected membrane lipids. They demonstrated potent antimicrobial activity, particularly newlineagainst Staphylococcus aureus, a common diabetic wound pathogen. These formulations newlinealso displayed anti-inflammatory effects, inhibiting various enzymes and maintaining newlinemacrophage cell viability while reducing nitric oxide overproduction in stimulated cells. newlinePhase II confirmed the safety of formulations for dermal use. Phase III showed that newlineJT-AFI and JT-YG promoted cell proliferation, migration, and angiogenesis while newlineregulating the cell cycle and mitigating ROS generation under hyperglycemic conditions.