Experimental Study On Bone Tissue-engineeringseeded With Bone Marrow Stromal Cell(BMSC)

Bone repair remains a clinical challenge. With the development of tissueengineering, a more ideal technique for bone repair is possible. In application of bone tissue-engineering, the source of osteoblast is the first problem to be solved. Osteoblast from bone marrow is the optimal seed cell, but for the heterogeneity of bone stromal cell (BMSC), the culture condition for osteogenic transformation of BMSC was to be improved ; biocompatibility and growth of cells on artificial carrier, the osteogenic potential of adult BMSC to be further researched. In this study,improvement had been made in the culture of BMSC for osteogenic transformation, cells seeded on hydroxyapatite (HA) and osteogenic potential of adult BMSC were investigated more in details.1 .Improvement on culture condition for osteogenic transformation of BMSCBMSC were derived from adult patients, the passage were divided into 4 groups: dexamethathone group, Danshen group, dexamethathone and Danshen group, control group. Cells were observed under invert microscope and growth curve established respectively. Osteogenic transformation rate and cell division rate were calculated. The result showed that Danshen cloud significantly eliminated the inhibitive effect of dexamethathone on proliferation of BMSC, co-culture by dexamethathone and Danshen could improve the osteogenic transformation dramatically.2. Study on ostoegenic potential of adult BMSCBone marrow were obtained from 13 patients, the stromal cells were cultured in 1640 medium containing dexamethathone (10^-8 mol/L), B-glycerophosphate(10mmol/L) and Vitamin C (50ug/ml). Cells were observed under invert microscope, osteogenic transformation rate was calculated and alkaline Phosphatase activity was examined .Calcium and bone matrix stains were used to relieved the new mineralized bone tissue . BMSC from 13 patients were cultured successfully, the osteoblast-like cells from BMSC expressed high alkaline Phosphatase activity. The node was proved to be bone tissue. It suggested that osteoblast-like cells from BMSC of adult had high osteogenic potential and could form mineralized bone tissue in vivo.3.Experimental study on application of BMSC in bone tissue-engineeringBMSC from rabbit were cultured in 1640 medium and 1640 medium containing dexamethathone (10^-8mol/L), B-glycerophosphate(10mmol/L). The passage cells were seeded onto blocks of hydroxyapatite (HA), cells were observed under invert microscope and scanning electron microscope; cells and HA were implanted into rabbit back muscle. After four weeks, block of HA were examined histopathologically. Cells were attached to HA and grew favourably, new bone were found in HA block, but fibrous tissue also observed in HA seeded with BMSC cultured in 1640 medium.The result show that BMSC were the optimal seeded cells in bone tissue-engineering, especially after conditioned culture, and HA was a suitable carrier for the seeded cells.