Adipose-derived Stem Cells Combined With Platelet-rich Plasma Promote The Healing Of Diabetic Wounds

OBJECTIVE: To observe the effect of adipose-derived stem cells(ADSC)combined with platelet-rich plasma(PRP)gel on wound healing and to explore its mechanism.METHODS:(1)ADSC of male SD rats were isolated and cultured by enzymatic digestion;PRP was prepared by Sonnleitner method;female diabetic SD rats were modeled by high-fat,high-sugar diet and 1% intraperitoneal injection of streptozotocin;(2)A wound model was constructed in diabetic rats and wounds were treated with PBS,PRP,ADSC,and ADSC+PRP,respectively.The growth of granulation tissue was observed on the 1st,3rd,7th,10 th,and 14 th day after operation.The wound tissue was embedded with paraffin and sliced.HE staining,immunohistochemistry,and immunofluorescence were used to observe the vascularization,Re-epithelization,macrophage infiltration,epithelial-mesenchymal transition and scar formation of the wounds;(3)MTS assay,flow cytometry,and in situ hybridization were used to observe the proliferation of endothelial cells(EC)and migration of fibroblasts(3T3)cultured in PRP and ADSC media(ADSC-CM),as well as the effects of PRP promoting ADSC survival,outcomes,and function maintenance;(4)Immunohistochemistry and Q-PCR techniques were used to investigate PRP,ADSC,and ADSC combined with PRP on the expression of wounds healing-related molecules SDF1/CXCR7.RESULTS:(1)ADSC combined with PRP could significantly promote vascularization,re-epithelization,collagen maturation,extracellular matrix deposition,and reduce macrophage infiltration during the process of diabetic wounds healing(P<0.05);(2)ADSC and PRP could significantly enhance the proliferation of EC,the migration of 3T3,the expression of the wound healing-related molecules SDF1/CXCR7 as well(P<0.05).PRP significantly could heighten the proliferation of ADSC,decrease the apoptosis and necrosis(P<0.05),meanwhile,improve the survival and prognosis of ADSC in diabetic wounds via re-distribution of ADSC to structures such as para-vascular and epithelial tissues.CONCLUSIONS:(1)ADSC combined with PRP therapy promotes angiogenesis,re-epithelization,inflammation,epithelial-mesenchymal transition,reduces scar formation in diabetic wounds,thereby accelerating the healing of diabetic wounds;(2)ADSC and PRP promote the process of diabetic wounds healing,enhance the proliferation of EC,the migration of 3T3,the expression of SDF1/CXCR7,a wound healing-related molecule,as well;(3)PRP can promote the proliferation of in a high-sugar environment and reduce its apoptosis and necrosis.ADSC participate in skin and vascular tissue structure remodeling to accelerate the healing of diabetic wounds.