The Study About Effect Of Adipose-Derived Stem Cells On Hypertrophic Scar Through TGF-β₁/Smads Signal Path Way In Paracrine Effect

Objective: To explored the feasibility of applying human adipose-derived stem cells(ADSCs) paracrine action to effect the Hypertrophic Scar(HS) fibroblasts(Fbs),and to investigate the cellular and molecular mechanisms base on TGF-β1/Smads path way and CollagenⅠ/Ⅲ. Methods: Human lipoaspirates was obtained from healthy patients who had recived abdominal liposuction.Then processed lipoaspirates cells were isolated and cultured as following described. Hypertrophic Scar(HS) tissue fibroblast(Fbs)and normal skin tissue dermal fibroblast isolation and culture proformed as described in artical.To verify the effect of soluble factors secreted by ADSCs on Hypertrophic Scar(HS) tissue fibroblast(Fbs). Hypertrophic Scar tissue fibroblast(HSFB) were cocultured with ADSCs in transwell chambers with a 0.4-um pore size membrane(experimental group). Normal skin tissue Fbs and Hypertrophic Scar tissue fibroblast(HSFB) were cultured alone as control group. Cmpare the morphology of fibroblast from normal skin and hyperplastic scar and coculture with ADSCs under inverted phase-contrast microscope. Proformed proliferation assay with CCK-8 on three groups Fbs,then base on the OD450 draw growth curve. Intracellular proteins were collected for Western blot of TGF-β1 and P-Smad2/3 and Smad7 and CollagenⅠ/Ⅲ.Results:(1)Cocultured ADSCs reduced growth curve of HSFB compare with culture HSFB alone.Cocultured ADSCs group was significantly lower than HSFB alone group after 4 day(P<0.05). However experimental group was still higher than normal skin dermal fibroblast growth curve and the difference was reducing at the begining of 3day. Meanwhile,no statistical difference was detected between coculture group and normal skin dermal fibroblastat the begining of 5day.(2) Coculturedexperimental group protein TGF-β1/P-Smad2/3/COLⅠ/Ⅲexpression level lower than HSFB culture alone group.And the difference have statistical difference between the two group showed time-dependent. But cocultured experimental group protein of TGF-β1/P-Smad2/3and COLⅠ/Ⅲ expression level still higher than normal skin dermal fibroblast,and have statistical difference. Smad7 protein expression level of experimental group were higher than the two control groups.Conclusions: Through the study on interaction between ADSCs and HSFB in coculture system, it was shown that ADSCs reduced HSFB proliferation ability in a paracrine manner.Cocultured ADSCs could decrease TGF-β1/P-Smad2/3/COLⅠ/Ⅲ protein expression level in TGF-β1/Smads path way, while promoted the inhibiting factor Smad7 expression level.