| It is difficult to cure the bone defect that is common in clinic field caused by many kinds of diseases. In the resent years, the development of bone tissue engineering has given us a new kind of thought and method, that includes three aspects: seed cell, growth factor and scaffolds. Now, the emphasis of life science is mesenchymal stem cell (MSCs) and its inductive factor. Basing on the clinic experience that Sheng-Ji Ointment can successfully cure infectious open fractures, the experiment tries to make Sheng-Ji liquor as a stimulative of mesenchymal stem cells.Objective: 1.to establish and perfect the technique of separation and purification of MSCs and to obtain MSCs maximally. 2.The Chinese traditional characteristic medicine , curing cure infectious open fractures ,is used to proliferation and differentiation of MSCs , in order to get a new kind of stimulative factor for bone tissue engineering. We can further have knowledge of the Chinese traditional medicine and put it to a brand-new application in bone tissue engineering.Method: l.Bone marrow is from rabbit tibia. MSCs are isolated by the method of density gradient centrifugation with stick to plastic, then, cultivated and identified. 2. We withdraw Sheng-Ji liquor and put it to MSCs in different concentration. The blank control is MEM medium without Sheng-Ji liquor, and the standard control is the same one with DEX, β -GP and VitC. We can observe the effect of Sheng-Ji liquor on MSCs by means of cell morphology and MTT evaluation. 3. We adopt positive control (cultured osteoblast) and two following ones to evaluate the osteogenesis of MSCs under the Sheng-Ji liquor in different concentration. Three are some ways: immunocytochemistry (for Col I ), acheomycin labeling (for mineralized nodes), detect complex (ALP/OCN/LDH) and calculate ratio (ALP /LDH, OCN / LDH).Result: 1 MSCs can be isolated by the method of density gradient centrifugation with stick to plastic. Its primary culture period is 10-11 days,... |
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