The gold standard for chronic wound treatment involves the grafting of skin tissue, which is currently becoming replaced by cellular therapy. Studies indicate that cellular functions occur via paracrine signaling factors rather than direct interactions. This finding has initiated the exploration of secretome-based therapy. Platelets and MSCs are two critical cell types involved in wound healing. In the present study, we focused on analyzing the effect of the MSC secretome and platelet-induced MSC secretome on wound healing. Adipose tissue-derived MSCs (ADMSCs) and PRP were isolated from healthy rabbits (New Zealand White, 2 kg, 6 months) after obtaining Institutional Animal Ethics Committee clearance (SCT/IAEC-439/July/2022/113). In order to prepare PRP-induced ADMSC secretome, ADMSCs were seeded in a 75 Cm² culture flask at a density of 15000 cells/cm². At 70% confluency, the serum-containing culture medium was replaced with 450 µl PRP (350-400 x10³cells/µl)-containing media and cells were incubated for 48 hrs. Hyperglycemia was induced by growing fibroblasts in high-glucose (30mM) DMEM with 5% serum and antibiotics. The wound-healing efficiency of PRP-induced MSC secretome and uninduced MSC secretome was evaluated by means of on-cell proliferation, migration, collagen synthesis, ROS generation, actin cytoskeleton deposition, and gene expression analysis. All quantitative data were expressed as mean ± SD (n=3). Results were analyzed using one-way ANOVA/Student's t test and were considered significant when *p ≤ 0.05.

We observed that PRP-induced MSC secretome treatment significantly increased cell proliferation and migration, improved collagen synthesis, reduced ROS generation, and promoted actin deposition in hyperglycemic fibroblast cells compared to uninduced secretome treatment. In conclusion, these findings suggest that PRP-induced MSC-conditioned media may be developed as an off-the-shelf therapeutic aid. Further in vivo evaluation of PRP-induced MSC secretome needs to be performed in diabetic models, and the molecular pathways underlying its mechanism of action also need to be explored as future research perspectives.