Therapeutic Effect And Mechanism Of Umbilical Cord Mesenchymal Stem Cells On Diabetic Foot Ulcer

BackgroundDiabetes is a common metabolic disease,which can cause peripheral neuropathy and peripheral arterial occlusive disease(PAOD),cause recurrent infection and ulcer of lower limbs,and cause diabetic foot ulcer(DFU).This kind of delayed wound healing caused by ischemia,infection,neuropathy and metabolic disorder poses a severe challenge to patients and clinicians.In recent years,with the changes of global nutrition and lifestyle,the prevalence of DFU has increased dramatically,but conventional treatment methods can not improve the status of high amputation rate and high recurrence rate of DFU.In recent years,mesenchymal stem cells(MSCs)are considered as an effective clinical treatment for DFU because of their strong ability of promoting vascular regeneration,immune regulation and transdifferentiation.Objective1.The clinical effect of UCMSCs on diabetic foot ulcer.2.Explore the potential mechanism of ucmscs in the treatment of DFU.Methods1.UCMSCs were separated and extracted by enzymolysis,the morphology of UCMSCs was observed,and the surface markers of UCMSCs were detected by flow cytometry.MTT method was used to detect the cell activity of UCMSCs under different generations and different inoculation densities,and the differentiation of UCMSCs into adipogenic,osteogenic and chondrogenic lines was carried out in vitro,and the differentiation results were identified by oil red O,alizarin red and alcian blue staining.2.The expression of VEGF,bFGF,Ang-1 and HGF in UCMSCs was identified by qPCR and ELISA at the transcription and expression levels respectively.PBMCs were isolated and co cultured to observe the chemotaxis of PBMCs to UCMSCs and the secretion of inflammatory and anti-inflammatory factors.3.Define the condition of DFU patients and observe the treatment of UCMSCs in two DFU patients.Patient 1: inject UCMSCs from day0 to day18 every three days,from near to far.Observe the wound healing in day-3,day0,Day6,Day12,day21 and day27,respectively.Routine blood test was carried out on Day10 and day25.Liver and kidney function test were carried out on day-1 and day25.Day32,day40,day46,day54 and day62 and day70 were followed up.Patients 2: day 0,day 3,day 6,day 9 and day 15 were injected with UCMSCs;day 17,day 18 and day 26 were followed up.Results1.The results of flow cytometry showed that UCMSCs highly expressed CD29,CD44,CD73,CD90,CD105 and HLA-ABC,but not CD34,CD45 and HLA-DR;oil red O,alizarin red and alcian blue staining showed that UCMSCs could differentiate into adipose,osteogenic and chondrogenic three lines;MTT showed that UCMSCs were alive in four generations There was no significant difference in sex.2.The results of qPCR and ELISA showed that UCMSCs could secrete VEGF,bFGF,Ang-1,IL-4,IL-10,TGF-? and other anti-inflammatory factors.The co culture system of UCMSCs and PBMCs showed that UCMSCs could collect PBMCs in 12 hours,promote the expression of anti-inflammatory factors and inhibit the expression of inflammatory factors.3.The patient was a typical severe DFU patient.After multiple UCMSCs injections on the foot surface,the wound healed and ulcer improved gradually within 30 days,and the condition was further relieved in the follow-up of one month.Blood routine test andliver and kidney function test showed that UCMSCs did not cause hepatorenal toxicity in patients with DFU.ConclusionUCMSCs can secrete many kinds of promoting blood vessel regeneration,UCM SCs can promote wound healing in DFU patients without hepatorenal toxicity.