ABSTRACT
BACKGROUND:Incidence of diabetic lower extremity vascular disease is increasing, so how to improve blood vessels of the lower limbs and increase angiogenesis becomes the research focus. Umbilical cord mesenchymal stem cells have been employed clinical y via local intramuscular injection, but the specific therapeutic effect and mechanism are not clear. OBJECTIVE:To investigate the effects of hypoxic preconditioning and cobalt chloride medium on the differentiation of umbilical cord mesenchymal stem cells into vascular endothelial-like cells in vitro. METHODS:Umbilical cord mesenchymal stem cells were isolated and cultured, and then treated with different concentrations of cobalt chloride. Enzyme linked immunosorbent assay was used to detect levels of basic fibroblast growth factor and vascular endothelial growth factor gene in cellsupernatants, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide to detect cellproliferation. The safety of umbilical cord mesenchymal stem cells before and after cobalt chloride induction was detected using chromosome. The umbilical cord mesenchymal stem cells were cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10μg/L vascular endothelial growth factor, 10μg/L basic fibroblast growth factor and 10%fetal bovine serum, which were induced to differentiate into vascular endothelial-like cells. Endothelial-like cellphenotype CD31 and von Wil ebrand factor were identified before and after induction, through the observation of three-dimensional model of angiogenesis, the angiogenic capacity of umbilical cord mesenchymal stem cells was determined. RESULTS AND CONCLUSION:Umbilical cord mesenchymal stem cells strongly expressed the surface marks. After the cobalt chloride induction, the proliferation of umbilical cord mesenchymal stem cells was positivelycorrelated with the time of induction. Based on the levels of vascular endothelial growth factor and basic fibroblast growth factor, the optimal concentration of cobalt chloride was 200 μmol/L. Chromosome detection showed the stability of cells after cobalt chloride induction. After induction, CD31 and von Wil ebrand factor were strongly expressed. Three-dimensional observation showed umbilical cord mesenchymal stem cells could be induced to form the lumen-like structure with different diameter sizes, and umbilical cord mesenchymal stem cells could be induced to differentiate into endothelial-like cells, and have a angiogenic capacity. RESULTS AND CONCLUSION:Umbilical cord mesenchymal stem cells strongly expressed the surface marks. After the cobalt chloride induction, the proliferation of umbilical cord mesenchymal stem cells was positivelycorrelated with the time of induction. Based on the levels of vascular endothelial growth factor and basic fibroblast growth factor, the optimal concentration of cobalt chloride was 200 μmol/L. Chromosome detection showed the stability of cells after cobalt chloride induction. After induction, CD31 and von Wil ebrand factor were strongly expressed. Three-dimensional observation showed umbilical cord mesenchymal stem cells could be induced to form the lumen-like structure with different diameter sizes, and umbilical cord mesenchymal stem cells could be induced to differentiate into endothelial-like cells, and have a angiogenic capacity.
ABSTRACT
Objective To analyse the risk factors relating to diabetic peripheral neuropathy(DPN).Methods 214 type 2 DM cases were investigated and the factors relating to DPN were analysed including age 、diabetic duration 、fasting blood glucose(FBG) 、post blood glucose(PBG)、 HbA_1c、blood pressure(BP)、 urinary albumin excretory rate(UAER)and lipid.Results Logistic regresstion analysis showed that UAER and FBG were relatively independent risk factors.Conclusion In many risk factors UAER and FBG are the most important factors of all in developling DPN.