Experimental Study Of Vascularization Of Tissue Engineering Skin Promoting By Erythropoietin Contrast Granulocyte Colony-stimulating Factor Combine Simvastatin

Objective:To observe the effect of erythropoietin and granulocyte colony-stimulating factor combine Simvastatin on the mobilization of EPCs (endothelial progenitor cells) in the bone marrow and vasculorization of tissue engineering skin(TES).Methods:Phase Ⅰ:According to the literature described dose (Simvastatin dose of2mg/Kg*d-1G-CSF dose of100ug/Kg*d-1EPO dose of3000IU/Kg*d-1) and methods, EPCs was mobilized by Recombinant Human Erythropoietin injection and granulocyte colony-stimulating factor combine Simvastatin in mice.108healthy male Kunming mice(avoiding the interference of estrogen) were randomly divided into EPO group, combine group and (NS control group), each group was36mice.6mice was randomly selected in each group after mobilization1d,3d,5d,7d10d,14d, Flk-1, CD34and CD133positive cell rate of selected mice heart blood was detected by FCM(flow cytometry), the results are statistics compared and analyzed. Phase Ⅱ:36healthy male BALB/C nude mice(avoiding the interference of estrogen and immunological rejection) were randomly divided into EPO group, combine group and NS control group, each group was12mice and continually mobilized for7d. Each group was divided into two subgroups:6mice were transplanted TES subcutaneously at5d and the other6were transplanted at7d. Mice were sacrificed14d after transplant. TES was then excised. The pathological morphology and vasculorization of TES was observed. The results, said as x±s, are Statistics compared and analyzed.Results:Phase Ⅰ:Double fluorescent positive cells of NS control group were not significantly different in6time point (P>0.05).The double fluorescent positive cells of EPO group and combine group was gradually increased and peaked at7d. From3d, in each time point the proportion of double fluorescence positive cells of threegroups have significant differences (P<0.05); double fluorescent positive cells of EPO group, compared with combine group and control group, increased significantly (P<0.05). Phase Ⅱ:vasculorization of tissue engineering skin among each group were significantly different (P<0.05). EPO group was better vascularized than combine group, but not vessels was observed in control group.5d transplanted group was better vascularized than5d transplanted group (P<0.01).Conclusion:The mobilization effect of EPO is better than granulocyte colony-stimulating factor combine Simvastatin. But Compared7d transplanted group,5d transplanted group is better in promoting vasculorization of transplat ed TES. Thus the time of transplanted TES should be at mobilized5d, when before the time EPCs to reached the peak.