Autologous Transplantation Of Mesenchymal Stem Cells As An Effective Therapeutic Approach For Lower Extremities Ischemia

basic research:Abstract: Mesenchymal stem cells (MSCs) possess of multi-potential to differentiate to various mesenchyma, such as vascular endothelial cells, which implies the possible effect of MSC to manage the peripheral artery disease. Vascular endothelial growth factor₁₆₅ (VEGF₁₆₅) is known to be intimately correlative with the growth of vascular endothelial cells. In the present study, we aimed to investigate the effect of transplantation MSCs with or without VEGF₁₆₅ to repair an ischemically damaged bind limb in a rabbit model. Autologous MSCs were condensatd by a separator from bone marrow samples taken from the tibia and cultured in DMEM preparing for transplantation or transfection. MSCs were transfected with non-replicative adenoviral vector with or without VEGF gene (hVEGF⁺ or hVEGF₁₆₅^-).Hind limb ischemia was created in 26 New Zealand White rabbits by means of rightfemoral artery ligation. Animals were randomized to receive intramuscular injection of either Dulbeco's Modified Eagle Medium (DMEM, groupl), hVEGFi65 (group 2) or hVEGF+165(group 3). Four weeks after treatment, ECT assessed perfusion in the damaged limb. The animals were euthanized and cells was harvested for real-time PCR and Western blotting to testify the results of transfection. After transplantation, it survived 5, 6, 8 rabbits in group 1,2 and 3 respectively. It was also confirmed that VEGF was successfully transfected into MSCs. ECT assessments revealed blood perfusion have increased at 2weeks after treatment in all three groups and the enhancing trend lasted to 4weeks in group2 or 3. In contrast to groupl, there is a significant increasing blood perfusion in group2 (at 2 and 3 weeks respectively, P<0.05) or group3 (at 1,3, 4 weeks respectively, P<0.05), but we failed to find statistic difference between group2 and group 3. In a conclusion, MSCs autotransplantation for limb ischaemia in rabbit model is an effective approach to enhance the peripheral perfusion. How to improve the effect of MSCs transplantation are still needed further investigations.Clinic research:Abstract: limb ischaemia is a common disease in old patients and presents an increase of incidence recent years. Conventional treatments, including medical agents and surgical operation are of limited efficacy,especially for critical ischaemic patients. For the shortage of effective treatments, surgeons always have to face the discomfiture of amputation for those patients. Searching for a novel therapy to help Regenerating a new vessel with less relapse is still the main aim. Mesenchymal stem cells (MSCs) possess of multi-potential to differentiate to various mesenchyma, such as vascular endothelial cells, which implies the potential novel approach to manage the peripheral artery disease. In the present report, we assess the efficacy and safety of low limb ischaemia treatment with MSCs autotransplantation.13 patients were involved in the study, which have been testified with low limb ischaemia by angiography and were evaluated to be unfit for angio-bridge operation. Autologous MSCs were condensatd by a separator from bone marrow samples taken from the iliac crest and cultured in DMEM preparing for transplantation. The cells were confirmed as MSCs by flow cytometry to test CD 117, CD29, CD166, CD45 and CD34 and then delivered intravascularly or intramuscularly. Before and after transplantation, the clinical parameters including still pain, chilly feeling and intermittent claudication in trouble limbs were graded. It was evaluated as following: healing as clinical symptoms disappeared;prominent improving as the symptoms straightened 2-3 grades;improving as 1 grade straightened and no efficacy with no improving. As a result, the alleviative rate in foot pain or cms pain is 80% or 100%, irrespectively. The reservation rate for limbsachieved 90%. 85% trouble limbs improved chilly symptom, and 90.9% ulcers in low limbs were healing. In addition, it helps to improve the effect of transfer-flow operation after transplantation MSCs in two patients. In a conclusion, MSCs autotransplantation for low limb ischaemia seems to be an effective approach to enhance the peripheral perfusion. Further investigations are still needed to elucidate the underlying mechanisms and long-term effect in this new treatment.