Effects of transplanted adipose derived stem cells on the expressions of α-SMA and DCN in fibroblasts of hypertrophic scar tissues in rabbit ears.

Abstract

To study the effects of transplanted adipose derived stem cells (ADSCs) on the expressions of α-smooth muscle actin (α-SMA) and decorin (DCN) in fibroblasts of hypertrophic scar tissues in rabbit ears. Twelve New Zealand white rabbits were selected; the normal subcutaneous adipose tissues in inguinal region were removed, ADSCs were extracted via enzyme digestion, cultured in Dulbecco's modified Eagle's medium (DMEM) and inoculated into the culture dish (3–5×104 cells/ml). After the rabbit ear hypertrophic scar model was established successfully, the fibroblasts of hypertrophic scar tissues in rabbit ears were separated and cultured using the mechanical method combined with enzyme digestion, and the ADSCs and scar fibroblasts were cultured in non-contact Transwell co-culture system for 21 days (experimental group); the corresponding scar fibroblasts were cultured in an ordinary 6-well plate without any treatment for 21 days (control group). The content of collagen I in fibroblasts was detected using the enzyme-linked immunosorbent assay (ELISA) kit, the mRNA expressions of α-SMA and DCN were detected via reverse transcription-polymerase chain reaction (RT-PCR), the protein expressions of α-SMA and DCN were detected via western blot analysis, and the expressions and distribution of α-SMA and DCN were detected via immunofluorescence assay. The results of ELISA showed that the content of collagen I in experimental group was decreased significantly (p<0.01). The results of RT-PCR and western blot analysis revealed that the mRNA and protein expression levels of α-SMA were significantly decreased (P<0.01, but those of DCN were significantly increased (p<0.01). Moreover, the results of immunofluorescence assay showed that the expression of α-SMA in experimental group was significantly decreased, while the expression of DCN was significantly increased. ADSCs can inhibit the mRNA and protein expressions of α-SMA and promote the mRNA and protein expressions of DCN in in vitro culture system, and they are expected to be used in the prevention and treatment of pathological scars.