Investigation of genes and miRNAs associated with scarless healing in orbital adipose‐derived and limbal‐derived mesenchymal stem cells treated with vitamin C

Abstract

Aims/Purpose: In this study we aimed to investigate miRNAs associated with scarless wound healing in limbal‐derived stomal‐mesenchymal stem cells (hLMSC) and orbital adipose tissue‐derived stem cells (hOA‐MSC) treated individually with vitamin C and exosomes obtained after vitamin C treatment.Methods: Isolation and characterization of hLMSCs and hOA‐MSCs were performed. MTT assay was employed to determine the ideal non‐cytotoxic and proliferative concentration of vitamin C (vit C). The selected concentration was applied to the cell culture medium of hLMSCs and hOA‐MSCs (vit C‐treated) followed by procurement of secreted exosomes (vit C‐exo) and their characterization. These exosomes were then used to enrich cell culture media of naïve hLMSCs and hOA‐MSCs. miR29, miR107, miR155, and miR543 miRNAs and Transforming Growth Factor Beta 1 (TGFβ1), TGFβ3 were analysed by Real‐Time PCR in both vit C treated and vit C‐exo‐treated cells.Results: Vit C‐exo and vit C‐treated hLMSCs demonstrated slight reductions in TGFβ1 and TGFβ3 expressions compared to controls. In contrast, a modest increase in TGFβ1 and TGFβ3 levels was noted in hOA‐MSCs after vit‐C and vit C‐exo treatment. After vit C‐exo application, expression of miR155 and miR107 increased in hLMSCs and decreased in hOA‐MSCs. An opposite trend was observed with miR543 expression with an increase in hOA‐MSCs and a decrease in hLMSCs. miR29 associated with fibrosis was dramatically increased in hOA‐MSCs, but not in hLMSCs.Conclusions: In conclusion, we show that miRNA and RNA contents of treated mesenchymal stem cells may vary depending on the origin of tissue. These results suggest that vitamin C could promote a relatively more pronounced anti‐fibrotic character in hOA‐MSCs.The authors acknowledge the support from Eskisehir Osmangazi University, Scientific Research Projects (ESOGU‐BAP Grant ID: TOA‐2022‐2458).