Experimental Study Of Accelerating Cutaneous Wound Healing With Paracrine Of Muscle Stem Cells

ObjectiveThe purpose of this paper was to investigate the effects of muscle stem cell conditioned medium(MuSC-CM)on the proliferation,migration and differentiation of fibroblasts in vitro.Full-thickness wound model was made to detect the effect of MuSC-CM on wound healing in rats.MethodsMuscle stem cells were isolated from the soleus of New Zealand white rabbit according to the different time taken for the cells to adhere.MuSC-CM was prepared and concentrated by ultrafiltration centrifuge tube.Five different growth factors in 10-fold concentration MuSC-CM were detected by enzyme-linked immune sorbent assay(ELISA).Fibroblasts were divided into several groups,including control group which did not add MuSC-CM(N-CM),5-fold concentration MuSC-CM group(5-fold CM),10-fold concentration MuSC-CM group(10-fold CM)and 20-fold concentration MuSC-CM group(20-fold CM).CCK8 assay was performed to study the efforts of different concentrations of MuSC-CM on fibroblasts proliferation,the effort of MuSC-CM on migration of fibroblasts was determined by scratch assay and the efforts of MuSC-CM on fibroblast differentiation was detected by Western Blot and 3D Collagen gel contraction assay.Full-thickness wound were created in the dorsum of rats.The rats were randomly divided into three groups,including 10-fold CM,N-CM and control.10-fold CM group: 60?l 10-fold CM was intradermally injected in the wound.N-CM group: 60?l N-CM was intradermally injected into the wound.contorl group: The wound was dressed without any treatment.The photos of wound at 0 day and 14 day were taken respectively to measure the rates of wound healing.ResultsThe MuSCs were isolated from gastrocnemius muscle and Desmin had been detected from MuSCs.The contentrations of TGF-?1,IGF-1,PDGF-BB,VEGF and bFGF in 10-fold concentration MuSC-CM were(6.38 ± 0.57)ng/ml,(2.74 ± 0.32)ng/ml,(1.42 ± 0.15)ng/ml,(2.19 ± 0.32)ng/ml and(4.35 ± 0.51)ng/ml,respectively.CCK-8 assay and scratch assay indicated that MuSC-CM can increase the proliferation and migration of fibroblasts in a dose-independent manner,respectively.Western blot showed that the fibroblasts treated with MuSC-CM had a significant higher expression of ?–SMA than control.Moreover,the results of 3D Collagen gel contraction assay showed that MuSC-CM can accelerate the contraction of 3D Collagen gel.The wound areas in 10-fold CM group were significantly smaller than that in N-CM group and control group after two weeks.ConclusionsMuSC-CM can accelerate cutaneous wound healing by increasing the proliferation and migration of fibroblasts as well as promoting the differentiation of fibroblasts into myofibroblasts.