APPLICATION AND EVALUATION OF MESENCHYMAL STEM CELLS ON WOUND HEALING IN RAT
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Date
2023
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West Bengal University of Animal & Fishery Sciences, Kolkata
Abstract
The primary goal of this study is to assess the wound healing effect of mesenchymal
stem cells and condition media in xenogeneic transplantation on rat skin full thickness
excisional wounds.
MSCs derived from bovine umbilical cord were isolated and cultured using explant
and enzymatic methods. The MSC isolation technique was standardised using enzymatic
digestion, as was the explant method. Because of their migratory properties, MSCs emerged
from explants by day 4 and reached 70-90% confluence by day 10. MSCs were digested with
collagenase (1mg/ml) for 4 hours at 370C before being plated and cultured, reaching 90%
confluence after 72 hours. MSCs exhibited strong plastic adherence and fibroblast-like
morphology. MSCs were cultured in vitro and differentiated into osteocytes, chondrocytes,
and adipocytes according to International Society for Cell Therapy and Mesenchymal Stem
Cell Committee guidelines.
Xenogeneic bovine UC-MSC and bovine UC-MSC derived condition media were
transplanted into two treatment groups and compared to positive, negative (vehicle), and
wound healing ointment groups. A full thickness (10mm) skin punch biopsy was performed
on both sides of the shoulder region, and MSC and CM were injected at the wound's
periphery. Wound healing was evaluated using wound contraction (up to day 14), gross
morphological evaluation (up to day 7), cytological examination of the wound bed area (up to
day 7), and histo-morphological evaluation of skin samples collected on days 7, 14, 21, and
28. Daily photographs were taken from the same height, and wound closure was accelerated
in the MSC and CM treated groups. Gross morphological parameters were scored, and it was
discovered that all wounds in all experimental groups were grossly healthy until day 7, with
enhanced granulation tissue noted in MSC and CM treated groups. Cytological analysis
revealed a significant increase in mononuclear infiltration after 72-96 hours in the CM and
MSC groups. Overall, histomorphological observations revealed that MSC and CM treated
groups had significantly accelerated wound closure, advanced re-epithelisation and neovascularisation,
mature extra-cellular matrix deposition with better collagen arrangement, and
reduced scarring. Every parameter was scored and statistically evaluated.