Establishment And Evaluation Of Autologous Transplantation Of Bone Marrow Mesenchymal Stem Cells On Diabetic Foot

Aims:To establish and evaluate the efficacy and safety of autologous transplantation of bone marrow mesenchymal stem cells on patients with diabetic foot.Methods:Forty five patients of Type 2 diabetes with critical limb ischemia were enrolled and randomized to either transplant group or control group. Patients in both group received the same conventional treatment. Meanwhile, 20ml bone marrow from each patient in transplant group was collected,and the mesenchymal stem cells were separated by density gradient centrifugation and cultured in the medium with autologous serum. Three weeks'adherent culture later, mesenchymal stem cells were harvested, identified by flow cytometry and detected by the experiment of athymic mouse transplantation tumo, chromosome analysis and pathogens culture. After that, mesenchymal stem cells were transplanted by multiple intramuscular and hypodermic injections into the impaired lower limbs. The follow-up indices include rest pain, painless walking distance, ankle-brachial index, magnetic resonance angiography, ulcer healing rate and amputation rate.At the same time, specific attention was paid to any potential complication or adverse effects resulted from the treatmen during follow-up.Results:The cells were in fusiform shape, aligned in parallel or whirlpool and exhibited the characteristics of human MSCs, i.e. being CD34,CD45-negative and CD44, CD73, CD90, CD105 -positive. The cells growed at logarithmic growth rate,and the numbers of these cells reached 7.32×108～5.61×109 after 3 weeks culture.There was no tumor tissue growing in the experiment of athymic mouse transplantation tumor.Chromosome analysis of the cells were normal and no pathogens were detected. At the end of 12-weeks follow-up, main ischemic symptoms,including rest pain and intermittent claudication,improved significantly in transplanted patients,and the mean of rest ABI in transplanted group significantly increased (P<0.001). All difference-mean of the above index in transplant group was significantly higher than that in the control group (P<0.001).The ulcer healing rate of the transplanted group was significantly higher than the rate of the control group(P=0.012).In the transplant patients, the rate of the patiants whose score of new vessels exceed or equal 2 in analysis of magnetic resonance angiography (MRA), was significantly higher than that in the control patients (P=0.001). Lower limb amputation rate in transplanted group was significantly lower than the rate in the control group(P =0.040). Neither complication nor adverse effects were observed as a result of the treatment.Conclusions:These results indicated that the autologous transplantation of bone marrow mesenchymal stem cells relieve critical limb ischemia, increase collateral circulation,promote ulcers healing,reduce amputation rate in Type 2 diabetic patients, and is a simple, safe, and effective therapy for diabetic foot.