Topical gel of mesenchymal stem cells-conditioned medium under TNF-α precondition accelerates wound closure healing in full-thickness skin defect animal model

Abstract

Mesenchymal Stem Cells (MSCs) under TNF-α stimulation (MSC-CM-T) can release numerous trophic and survival molecules that have a promising prospect in wound healing acceleration. However, the effective levels of MSC-CM-T in topical gel preparation to accelerate wound healing should be further explored. The aim of this study was to investigate the effects of MSC-CM-T in topical gel preparation in accelerating optimal wound healing through analyzing PDGF levels, wound closure rate percentages, and fibroblast density appearances. Twenty-four male Wistar rats were performed a full-thickness excision. The group studies were randomly assigned into four subgroups: control gel, control medium, and two treatment groups: MSC-CM-T topical gel at doses of 100 μL and 200 μL (T1 and T2, respectively). Wound closure rates were measured by standard caliper, platelet-derived growth factor (PDGF) levels were analyzed using ELISA on days 3 and 6, whereas the fibroblast density appearances were determined using hematoxylin-eosin staining. This study found a significant increase in PDGF levels in all treatment groups on days 3 and 6. These findings were in line with the increase of wound closure rates in all treatment groups on day 6, in which the high dose of MSC-CM-T was more effective in initiating the increase of wound closure rate. We also found the fibroblast density appearances on day 6 in the T2 group. We conclude that the topical gel of MSC-CM-T is more effective in accelerating wound closure healing through increasing PDGF levels and wound closure percentages and fibroblast density appearances in the skin defect animal models.

Keywords: ANOVA – Analysis of Variance; APC – Allophycocyanin; BSA – Bovine Serum Albumin; CD – Cluster of Differentiation; DMEM – Dulbecco’s Modified Eagle Medium; ELISA – Enzyme-linked Immunosorbent Assay; ERK – extracellular signal-regulated kinases; FBS – Fetal Bovine Serum; FITC – fluorescein isothiocyanate; Fibroblast; HE – Haematoxylin and Eosin; HLA-DR – Human Leukocyte Antigen-DR; ISCT – International Society of Cellular Therapy; LSD – Least Significant Difference; MAPK – mitogen-activated protein kinase; MSC-CM-T; MSC-CM-T – Mesenchymal Stem Cells-Conditioned Medium; MSCs; MSCs – Mesenchymal Stem Cells; PBS – Phosphate Buffer Saline; PDGF; PDGF – Platelet-derived Growth Factor; PerCP – peridinin chlorophyll protein; SD – Standard Deviation; T1 – Treatment 1; T2 – Treatment 2; TNF-α – Tumor Necrosis Factor-α; UC – Umbilical Cord; VEGF – Vascular Endothelial Growth factor; Veh – Vehicle Control; wound healing.