Effect Of Danhong Injection Combined With HADSCs On The Survival Rate Of Autologous Fat Transplantation

Objective: This study investigated the effect of Danhong injection(DH)combined with human adipose-derived stem cells(hADSCs)on the fat survival rate in fat-grafted animal models and found that the new method to improve the survival rate of autologous fat transplantation(AFT)is to achieve better autologous fat transplantation in clinic.Methods:1 The human adipose-derived stem cells(hADSCs)were isolated from specimens and culture in vitro,the cells' characteristics were detected by the related antigens expression of the third generation of human adipose-derived stem cells(hADSCs)were detected by flow cytometry.The third generation of hADSCs reserved spare.2 The prepared fat granules were divided into four groups according to different intervention facts,A:fat granules;B: fat granules+ hADSCs;C:Danhong injection+ hADSCs;D: fat granules+ hADSCs+ Danhong injection.The experimental nude mice were randomly divided into 6 groups with each group.Fetal fat particles were injected into the dorsal surface of nude mice to establish experimental model.2,AB groups of nude mice intraperitoneal injection of saline daily,CD groups of nude mice injected with the same amount of Danhong injection for intervention.At the 15 th day and the 30 th day after transplantation,and the grafts were obtained from 3 nude mice were sacrificed in each group.The graft volume was measured the fibrosis of the graft and the appearance of the capillaries were observed under a microscope the density of blood vessels was measured by immunohistochemical staining.Results:1 hADSCs were isolated,cultured and identified: the adipose tissue was treated with the cell suspension,then inoculation in a cell culture flask.A large number of spindle cells and a few polygonal cells were seen in the cultureflask after 7-9 days.Fourteen days or so visible spindle cells were spiral or bundle growth.Primary cells can reach 80% confluency for passage.The third generation of human adipose-derived stem cells(hADSCs)was detected by flow cytometry.the results showed that the positive rates of CD29,CD44 and CD90 were over 92%,while the positive rates of CD31,CD45 and CD146 were all less than 6%(Fig.5),almost not expressed,to prove that the cells cultured in this experiment is indeed human adipose-derived stem cells(hADSCs).2 Graft visualization: 15 d after transplantation,the graft surface formation of envelopes,nascent capillaries walking in the envelope surface,the results of the four groups were compared: Group D graft was the most fresh capillaries,A group at least,B,C two Group between A,D.30 d after transplantation,the volume of the four groups of grafts were compared: the volume value obtained by LSD-t method for any comparison: A and B,A and C,A and D,B and D,C and D(P <0.05).There was no significant difference between group B and group C(P> 0.05),and the volume of graft in group D was the largest,group A was the smallest,group B and C were both Group between A,D.3 The results of Immunohistochemical analysis of the section: 15 d after transplantation,immunohistochemical analysis of the four groups' fat graft immunohistochemical slice value LSD-t method for comparison,A and B,A and C,A and D,B and D(P <0.05).There was no significant difference between B and C(P> 0.05).The results showed that the microvessel density in group D was the highest.group A was the lowest group,B,C two groups between A,D group.Conclusion: The combination of DH and hADSCs with autologous fat grafts(AFT)was better than each single factor intervention in improving the survival rate of fat grafts in animal models,and the density of newborn capillaries around the grafts was even greater.