Effect Of Adipose Derived Stem Cells Combined With Platelet-rich Plasma On Wound Healing In Diabetic Rats And Its Mechanism

Objective:To asses the effects of combining adipose derived stem cells(ADSCs)with platelet rich platelet(PRP)on the repair of diabetic ulcer wound,to provide scientific foundations for the clinical application.Method : 55 clean grade male SD rats(body weight 180-220g)were prepared,6 were randomly selected for heart blood extraction to prepare PRP,4 were randomly selected for ADSCs extraction,and the remaining 45 were randomly divided into 5 groups with 9rats in each group.One healthy group was served as the normal control group.Diabetes was induced in the other four groups by intraperitoneal injection of streptozotocin(STZ)at40mg/kg after one week of adaptive feeding.After 3 days the random blood glucose was greater than 16.7mm,and after 1 week the rats with stable blood glucose were deemed to have successfully constructed the model.The 4 groups of rats were randomly divided into diabetes group,ADSCs group,PRP group and combined treatment group for the experiment.Ninety 1.5cm×1.5cm wounds were created in the symmetrical part of the back of 45 rats,and the healing of the wounds,the changes of skin cell pathology and related factors were observed on days 3,7 and 14,respectively.Result:With the prolongation of time,the wounds of the rats in each group healed to different degrees.Among them,the diabetic group had the slowest healing rats,with the normal group had the fastest healing rate.The combined treatment group had a faster healing rat than the ADSCs group and the PRP group.HE staining showed that compared with the diabetes group granulation tissue production was significantly increased in the combined treatment group and the number of inflammatory cells was decreased.Masson staining and Wolf scarlet staining showed that collagen fiber production in the combined treatment group was significantly better than in the diabetes group.The expression levels of collagen genes,TGF-?,Col1a1 and Vim were low in the diabetes group,and the combination treatment group can effectively increase the expression level of ?-sma in the diabetes group.The expression of CD31 in enthothelial cells in diabetic group was lower than that in combination treatment group,while the expression in non-endothelial cells was higher than that in combination treatment group.VEGF expression reduced in the diabetic group and combination therapy could change this.The expression of apoptosis-related genes caspase-3 and 9 was higher in the diabetic group,the expression of cyclinD1 and Pcna was lower,and the expression of mmp-2 and 9 was higher,which was not conductive to wound healing.The combination therapy group was able to change the expression of these genes.Conclusion:The combined application of ADSCs and PRP can reduce the level of wound cell apoptosis,promote cell proliferation,and reduce the content of MMPs,thus promoting the generation of wound granulation tissue and collagen,and ultimately promoting wound healing in diabetic rats.