| Type 2 diabetes mellitus (T2D) is a metabolic disorder characterized by hyperglycemia. The hyperglycemia of diabetes mellitus may cause internal metabolic disturbance, destroying bone, and affecting the process of the deposition of sclerotin, bone remodeling, and bone mineralization. Diabetic patients are vulnerable to refractory fracture, protracted course of surgical incision healing, periodontitis, microvascular complications, etc. in clinical treatment.Bone marrow mesenchymal stem cells (BMSCs) is the kind of cells with the capacity of multi-differentiation potential, which is important seed cells in the healing process of damaged bone tissue. The healing of bone tissue relies not only on the combined action of osteoblasts in the damaged part and BMSCs, etc., but also on the migration and the osteogenic differentiation of neighboring tissue and BMSCs of the whole body. Therefore, how to implant the migration of the BMSCs of diabetic patients to the damaged part for osteogenic differentiation is one of the hot issues of bone tissue regeneration. Studies show that the damaged tissue formed an endogenic biological electric field at the wound site, and the wound could hardly recover without the regulating effect of such field. Previous experiments demonstrated that Direct current electric field (DCEF) could effectively promote normal BMSCs migration and osteogenic differentiation ability.In this study, GK rat with T2D is used to test and screen the suitable DCEF intensity and exposure time that promotes the migration of diabetic rat BMSCs and osteogenic differentiation, to provide a theoretical and experimental foundation for a new strategy for improving the success rate of damaged tissue repairing of diabetics.Aim:1. The influrnce of DCEF on rBMSCs of GK rat with T2D was studied.2. The intensity and exposure time of DCEF that enhance the migration ability of rBMSCs were screened using the transwell chamber method.3. The ostrogenic genes and proteins were evaluated, thus to determine the influences of DCEF on osteogenic differentiation of rBMSCs.Method:1. Made comparison of BMSCs biological behavior between Wistar rat and GK rat with T2D.2. Observed the BMSCs migration direction of Wistar rat and GK rat with T2D under a DCEF. Screened out the optimum field intensity and exposure time that could promote BMSCs migration of Wistar rat and GK rat with T2D, and compared the difference between the two kinds of cells under a DCEF.3. Preliminarily discussed the effect of the optimum DCEF on the BMSCs proliferation, osteogenesis-related genes, and protein level of GK rat with T2D.Result:1. The BMSCs of Wistar rat of the third generation appear of uniform fusiform, and had a strong ability of proliferatioin, and osteogenic and adipogenic differentiation; as to the BMSCs of GK rat with T2D, the viability was low, the cell morphology stretch was weak, the alignment was loose, and some appear fusiform, and others diamond-shaped or polygonal; the osteogenic differentiation ability of GK rat with T2D was weaker than that of Wistar rat, but the adipogenic differentiation ability was slightly better. The flow cytometry test showed that the surface marker of CD44, CD 105 and CD29 cells is positive, while that of CD45 and CD34 cells was negative.2. The BMSCs of both Wistar rat and GK rat with T2D migrated to positive pole after being intervened under a DCEF for 4 hours. The amount of migrating BMSCs of both Wistar rat and GK rat with T2D reaches peak in the event that the intensity of DCEF reached 200mV/mm, and the exposure time reaches 4 hours. DCEF could effectivcely promote the BMSCs proliferation of GK rat with T2D.3. Under the optimum DCEF. both the ALP and RUNX-2 gene expression level and the protein expression level of BMSCs of GK rat with T2D were significantly improved.Conclusion:1. Comparing with Wistar rat. the BMSCs shape of GK rat is not regular, the BMSCs proliferation ability and the osteogenic capacity are low, but the adipogenic differentiation ability is slightly better; Tests with a flow cytometry show that both kinds of cells have the characteristics of stem cell, and multiple differentiation potential.2. The BMSCs of both Wistar rat and GK rat with T2D are of galvanotropism, and their orient migration direction is positive. The optimum condition for promoting the BMSCs migration, osteogenesis, and proliferation of GK rat with T2D is 4-hour exposure to 200mV/mm DCEF.3. DCEF can promote the BMSCs osteogenic differentiation of GK rat and Wistar rat increasing both osteogenic gene and protein expression. |
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