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Objective: Large mandibular defect repair has always been the difficulty of clinical, although many alternative treatments, but few have a way to compare the ideal solution to fix the problem, to obtain satisfactory results. Bone tissue engineering propose to solve this problem provides a new way. Osteoblasts as seed cells for bone tissue engineering, it is the acquisition and cultivation of tissue engineering is the most basic and the most important aspects. A variety of sources in osteoblasts, the bone marrow is most advantageous. Bone marrow-derived osteoblast cells from bone marrow progenitor cells from but limited number of bone progenitor cells, proliferation slow, difficult to adapt to the seed cell bank needs large number of cells. In this study, bone marrow stromal cells as a source of seed cells for tissue engineering Screening cultured rabbit osteoblasts and their biological characteristics observed for ways to get screened seed from the bone marrow cells and bone tissue engineering applied to provide a mandibular defect repair experimental basis; and explore the insulin-like growth factor Ⅱ (insulin-like growth factor Ⅱ, IGF Ⅱ), basic fibroblast growth factor (basic fibroblast growth factor, b-FGF), transforming growth factor-β (transforming growth factor β, TGF-β) of these three growth factors alone or in combination interact osteoblast proliferation after the impact, hoping to choose a suitable promote osteoblast proliferation approach for osteoblast bone tissue engineering to adapt to the library lot needs to provide some experimental basis. Methods: Japanese white rabbits extracted bone marrow fluid obtained after centrifugation of bone marrow mononuclear cells were cultured rabbit osteoblasts proliferation and growth factor in rats after washing with PBS to IX 10fynd cultured cell concentration, obtained after 2 weeks growth of adherent monolayer cells. Then enter the subculture through inverted microscope, HE staining, scanning electron microscopy, transmission electron microscopy, alkaline phosphatase (alkaline phosphatase, ALP) staining, I collagen immunohistochemical staining, calcium staining cells were obtained by means of biological characteristics research. The cells were then with the carrier material polylactic acid (PLA), expanded polytetrafluoroethylene (OPTFO composite culture one week after replantation of autologous rabbit muscle, four weeks, Pakistan weeks after implant sites drawn, histological methods in vivo osteogenic . and using cultured osteoblasts fifth generation, with the eagle Pi blue method, observation of insulin-like growth factor 11, basic fibroblast growth factor in transforming growth factor · p of these three growth factors in a separate and After the combined interaction of osteoblasts in vitro proliferation. Results: morphology, this experiment can be adherent cultured cells, showing a variety of forms, with cytoplasmic processes of uneven thickness, and showed a colony-like growth, may overlap into a complex layer growth and the formation of cell nodules electron microscope showed the typical structure of protein synthesis; cytoplasm rich in mitochondria, rough endoplasmic reticulum and the Golgi apparatus, indicating that the synthesis and secretion of functional strong, in line with the morphology of osteoblasts and growth characteristics in biochemical and \The continuous passage, form and function the same. osteoblast proliferation in vitro, co-culture with the carrier after growing well. implanted early after (4 weeks) bone matrix formed loose structure, the surrounding bone still more cells, late (8 weeks) the visible structure formed in dense bone tissue, bone cells within the lacuna in the bone, the bone tissue with the typical basic structure similar to the same culture conditions, all three growth factors can promote osteoblast proliferation, IGF.H, bFGF both growth factors in the concentration range in 0.l-10.ong/thl with rabbit osteoblast proliferation in a dose-effect relationship ·, and this concentration proliferation 10.onglml role in the most significant, concentration exceeds this value, but decreased proliferation effect. TGF-D at 0. olghl proliferation effect when the most significant, as the concentration increased, the declining role. choose better effect concentration of three kinds of growth factors in the rabbit osteoblasts cultured for 1 day, in addition to 10.0 il bFGF, the remaining pro-proliferative effect was not significant, three days after the first two rabbit osteoblasts with maintenance and growth factors in their proliferation in rats appear to promote proliferation differences, five days later, growth factors promote proliferation of rabbit osteoblasts reached the maximum seven days, weakened Three kinds of growth factor interactions when the rabbit osteoblast proliferation synergy, both to promote rabbit osteoblasts proliferation, and growth factors in bFGF + GF · 11 with bFGF + IGF · H can GF Little combined effect is more noticeable if Conclusion: two rabbit bone marrow stromal cells were cultured in vitro and actively growing, with osteoblasts similar morphology and growth Features; has strong ALP activity, can secrete type I collagen, to in vitro calcification; cells in a carrier complex can be implanted in the body eight weeks after the formation of bone tissue. showed that: We screened successfully cultured bone marrow cells are rabbits into bone cells, obtained from bone marrow osteoblasts is feasible. 2 three kinds of growth factors in bone marrow can promote osteoblast proliferation, growth factor promoting proliferation in a dose-dependent and time-dependent, three kinds growth factors have a synergistic effect when interaction, suggesting that growth factors can promote rabbit in vitro osteoblast proliferation, can be used as fast in a short time to get a seed cells for bone tissue engineering osteoblasts an effective way.
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