Bmsc Dbm Composite Repair Of Bone Defects And Benefit Kidney Soup Experimental Study Of Adjuvant Therapy

Objective: (1) To fractional cultivate and amplification the rabbits' bone marrow derived stroma cell in vitro. To observe the growth and biology features of primary and passage cells. (2) To observe the effect of Bushenyigu decoction (invigorating the kidney and tonifying bone decoction) on the BMS cells. (3) To construct recombination artificial bone and observe its' effect of repairing bone coloboma, compared with simple variant cancellated bone matrix repairing. (4) To observe the influence of Bushenyigu decoction in osteogenesis of recombination artificial bone by feeding traditional Chinese drug after operation.Methods: (l)Take suction marrow by bone puncture at double ilium with asepsis condition. BMS cells were separated and purificated by the method of density gradient separation. Trypsin digests and passages cells. The passage cells were cultivated by conditioned medium. To observe cell growth condition with inverted microscope everyday. Passage cells proliferation condition was detected by HE staining, quantitative assay alkaline phosphatase and MTT method. (2) The passage cells were inoculated in 24 holes cultivation plate (1.5×l0~4cells/ holes). The cells detected by enzyme detection machine after cultivated with traditional Chinese drug. BMS cells' proliferation condition influenced by Bushenyigu decoction was detected by MTT method (wavelength is 630nm). (3) Collect and cultivate BMS cells of healthy rabbits. Centrifuge (1500r/min, 10min) the cells and make them into cell suspension. Inoculate cell suspension in variant cancellated bone matrix and make tissue engineering artificial bone. The rabbits were divided into four groups (12/group) randomly. They were simple variant cancellated bone matrix group, recombination artificial bone group, recombination artificial bone group with feeding traditional Chinese drug and blank group. Make bone coloboma models. We observed and detected the normal condition, X-ray, ALP activity and histology every month (4) Traditional Chinese drug group injected Bushenyigu decoction by gastric canal (0.06g/kg) everyday. Tissue engineering artificial bone group injected distilled water the same as traditional Chinese drug group everyday.Results: (1) BMS cells cultured in vitro grow well. HE staining shows cleavage phases are more and the cells are fusiform shape. The result of ALP content show: it is (20.5 ± 1.29)U/ml in typical culture medium, (29.75 ± 4.85)U/ml in conditioned...