To Investigate The Effect Of Repairing Diabetic Wound By Rat Bone Marrow Mesenchymal Stem Cells On Modified Poly L Acid And Type Collagen I Composites Scafold

Objectiv:1. Culture and identify BMSCs in vitro,and to investigate the differentiation capacityof BMSCs；2. Made complex scafold by modified Poly L Acid and type collagen I，and tovestigate the feasibility of BMSCs differentiated into epidermal cells on PLA-I。3. Study on the effect of BMSCs by PLA-I on wound healing in diabetic rat。Methods:1. Culture and identify BMSCs in vitro by whole marrow method，Flow cytometricauthenticate the purification degree by detection the surface molecular markers ofBMSCs； detection osteogenesis, adipogenic and skin stem cells after addedbone-formation solution、adipogenic medium and induce solution of epidermal stem cell。2. Preparation PLA-I composite scafold, and detection skin stem cells by superficialstem cells markers keratin and integrin β1expression after added induced solution ofepidermal stem cell of the BMSCs on PLA-I composite scafold。3. The one-time intraperitoneal injection of STZ to establish the diabetic rats model；making Rats wound under anesthesia. labeled5-BrdU BMSCs were implanted atPLA-I composite scafold as tissue engineered skin and transplant to rat diabeticwound, observe wound healing, Use transparent membrane method for calculationand statistical analysis of the wound healing rate in different periods. take samplesof newly-grown skin to undergo HE staining and immunohistochemical in order toauthenticate the origin of epidermic cells. Results:1. BMSCs were isolated, purified and cultured successfully, Flow cytometricauthenticate the purification degree as high as94%, The successful Differentiationin vitro testing to cells calcification, fat particles and skin stem cells markerskeratin19and integrin β1.2. The successful Preparation PLA-I composite scafold, Add skin stem cells afterinducers, two-signed of immunohistoche-mistry identify CK19and β1was highlyexpressed at Rat BMSCs on PLA-I.3. The successful construction of the wound model of diabetes rat; Rat diabetic woundwere healing fast by transplanting5-BrdU BMSCs and PLA-I，HE staining showedthat newly-grown skin have more blood vessel、cutaneous appendages and thickerthen control group， the cytokeratin were detected in the5-BrdU BMSCs in thenewly-grown skin of the modeI rats，which is not founded in control rats throughdouble—labelling technique of immunofluorescence．Conclusion:1. whole marrow method can obtain high purity of BMSCs in vitro，it’s the top choice ofisolating culture in vitro on account of its simple and low demand. BMSCs have avariety of organs differentiation potential as osteogenesis, into adipocyte and skinstem cells.2. PLA-I have good trained relationship with BMSCs and do not affect its proliferationand differentiation ability.3. Rat diabetic wound were healing faster and better by transplanting BMSCs on PLA-I,diabetes refractory wound to provide,its a new tissue engineering skin treatments fordiabetic ulcers.