Gene and cellular assessment of wound healing with a novel natural cocktail gel dressing: A new method for quantitative wound closure time assessment.

Abstract

This study aimed to enhance wound healing using a novel natural cocktail gel dressing composed of purslane, human amniotic membrane (hAM), and platelet-rich plasma (PRP). In addition, a new ratio-based analytical approach was applied to evaluate the healing dynamics in each treatment group, revealing correlations between the healing rate and the respective treatment compound. Under aseptic conditions, specific amounts of freeze-dried decellularized hAM, PRP, and hydroalcoholic extract of purslane (HAEP) powder were prepared. The study groups included hAM gel, PRP gel, HAEP gel, a cocktail gel (HAEP + PRP + hAM), and phenytoin gel (positive control). Cytotoxicity was evaluated using the MTT assay. In vivo, seven groups were assessed on days 7, 14, and 21. Wound closure rate was analyzed via photographic imaging, and tissue samples were collected for H&E staining. Wound healing dynamics were further evaluated using 14/7-day and 21/14-day ratios. The cocktail gel significantly enhanced wound healing compared with other groups (p < 0.05), improving cell migration, M2 macrophage polarization, and angiogenesis. The ratio-based analysis indicated that the cocktail group exhibited the fastest healing between days 7 and 14, while purslane and hAM groups showed superior healing between days 14 and 21. This study introduces the use of 14/7 and 21/14-day ratios for the first time as a quantitative measure of healing progression, bridging macroscopic closure rates with underlying cellular and molecular changes. The combination of purslane, PRP, and hAM significantly accelerated healing and reduced closure time, suggesting a synergistic effect. The proposed ratio-based approach provides a more accurate evaluation of wound healing phases.