| Objective:The problems of autologous fat grafting are high absorption rate and low survival rate after autologous fat graft.In the process of biological outcomes after transplantation,the key to its survival rate is that improve fat cells the ischemia hypoxia tolerance ability,reduce host inflammatory response and fiber cystic change,enhance the anti-apoptosis capacity,set up timely and sufficient blood supply,angiogenesis.Thymosin beta 4 is the potential factor to promote the survival rate of free fat graft.its function include promoting angiogenesis and anti-fibrosis,anti-inflammatory and wound healing.According to its biological characteristics,it could be used in the fat transplantation that we explore its function and mechanism of action on the survival rate of fat cells and tolerance hypoxia ability.So we will provide theoretical and experimental basis for clinical application of free fat graft technology.Methods:We extracted adipose tissue from rabbit carotid back,and added T?4 with different concentrations in the free fat particles,then transplanted the adipose tissue to rabbit ears.The grafts in vivo at each time point were detected by MRI(magnetic resonance imaging).The fat grafts were harvested from each rabbit at each time point,then were evaluated general observation of adipocytes.Immunofluorescence staining of sections was performed that BODIPY-FL vital staining cell count and anti-CD31 antibody(a specific marker for endothelial cells).All the sections were imaged on fluorescence microscope.Results:1.The grafts in vivo at each time point(2,4,12w)were detected by MRI,then calculated the integrated optical density of enhanced scanning.The result showed that group A and group B of IOD scores significantly higher than that of group C(p<0.05),and comparison between groups had obvious difference(p<0.05).This suggested that thymosin beta4 improved microcirculation perfusion of grafts.2.In the study,the grafts were harvested from each rabbit at each time point(2,4,12w),measured the wet weight and volume after drainage method.Histological observation after HE staining was carried out on the survival rate of grafts.The result showed that group A and group B in adipose tissue integrity scored significantly higher than that of group C,inflammatory cell infiltration and fibrous connective tissue hyperplasia was significantly lower than that of group C(p<0.05).T?4 could significantly improve the survival rate of free fat tissue,and enhance lower rate of adipose tissue fibrosis.3.The group A and group B positive numbers of BODIPY-FL vital staining cells and lectin blood vessels at each time point((2,4,12w)were significantly higher than that of group C(p<0.05).The group B positive numbers of BODIPY-FL vital staining cells were significantly higher than that of group A(p<0.05).4.Immunofluorescence staining(anti-CD31 antibody on micro vessel-density counts)revealed that the number of the brown positive blood vessels at high magnification in group A and group B were significantly higher than that of group C(p<0.001).And three groups had statistically difference(p<0.05).Conclusion?This study has shown that T?4 could promote the survival and angiogenesis of fat grafts,then enhance the ischemic tolerance of adipocytes.thymosin ?4 is a potential factor that improving autologous free fat graft survival. |
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