[Effect of extracellular matrix and serum components on cellular adhesion and growth in vitro and in vivo].

K Nishimura
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Abstract

The recent cell-biological research have been reported the usefulness of extracellular matrix, such as collagen and fibronectin, in cell culture techniques and in vitro experiments. These matrix began to applied in medical treatments and brought many success. Fibroblast and osteoblast have characteristics to promote stabilization of implant materials in vivo. These cells revealed the additional effects to control mending and healing of surgical invasion. The role of extracellular matrix and serum components for cellular adhesion and growth have been discussed extensively. The recent studies reported that collagen and other materials promoted the cellular behavior. The author discussed the effect of extracellular matrix and serum components or the established cell lines on cellular adhesion and growth, which were performed in vitro and in vivo. The following cell lines were employed in this study, fibroblastic cell lines from mouse (3T12-3) and human (SF-TY), osteogenic cell line from mouse (MC3T3-E1) and osteogenic cell lines from human (SAOS-2, NY), osteoblastic cell line (HuO-3N1) and rabbit cell line. Different types of collagen were used as the coating material on glass surface by the method of collagen-gel culture method. Fetal calf serum, bovine serum albumin, fibronectin and proteoglycan were also employed in this study. After the cultivation, adherent cells were stained by Gimsa's method and counted microscopically. In cellular-adhesion test, the most cell number was found on the collagen type I coated glass surface to compare with the other materials, such as fetal calf serum, bovine serum albumin, proteoglycan, employed in all the tested cell lines. Growth of the defined cell-number on glass surface was compared among used materials, and all the osteogenic cell lines and fibroblastic cell lines were found to grow well on collagen type I-fibronectin, collagen type I, and fibronectin coated glass surface to compare with other materials employed in this study. Collagen-fibronectin and collagen-coated alumina ceramic tubes were embedded in the mandibular bones of rabbits, and the aspect was observed with the passage of time. These treatments promoted the osseous area in the tubes. These findings strongly indicated that surface-coated biomaterial by collagen and collagen-fibronectin enhanced cellular adhesion, and promoted the growth of fibroblast and osteoblast in vitro and in vivo.

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[细胞外基质和血清成分对体外和体内细胞粘附和生长的影响]。
近年来的细胞生物学研究报道了胶原蛋白和纤维连接蛋白等细胞外基质在细胞培养技术和体外实验中的应用。这些基质开始应用于医学治疗,并取得了许多成功。成纤维细胞和成骨细胞具有促进植入材料在体内稳定的特性。这些细胞显示了控制手术侵袭的修复和愈合的额外作用。细胞外基质和血清成分在细胞粘附和生长中的作用已被广泛讨论。最近的研究报道胶原蛋白和其他物质促进细胞行为。本文通过体外和体内实验,探讨了细胞外基质和血清成分或所建立的细胞系对细胞粘附和生长的影响。本研究采用的细胞系为小鼠(3T12-3)和人(SF-TY)成纤维细胞系,小鼠(MC3T3-E1)和人(SAOS-2, NY)成骨细胞系,成骨细胞系(uo - 3n1)和兔细胞系。采用胶原-凝胶培养法,将不同类型的胶原蛋白作为玻璃表面的涂层材料。胎牛血清、牛血清白蛋白、纤维连接蛋白和蛋白多糖也用于本研究。培养后,用Gimsa染色法对贴壁细胞进行染色,显微镜下计数。在细胞粘附试验中,I型胶原涂层玻璃表面的细胞数量最多,与所有细胞系中使用的胎牛血清、牛血清白蛋白、蛋白多糖等材料相比较。对比不同材料在玻璃表面的生长情况,发现所有成骨细胞系和成纤维细胞系在I型胶原纤维连接蛋白、I型胶原纤维连接蛋白和纤维连接蛋白涂层玻璃表面生长良好,与本研究中使用的其他材料相比。将胶原纤维连接蛋白和胶原包被氧化铝陶瓷管植入兔下颌骨,观察其随时间的变化。这些处理促进了管中的骨区。这些结果表明,胶原和胶原-纤维连接蛋白表面包被的生物材料增强了细胞黏附,促进了成纤维细胞和成骨细胞的体外和体内生长。
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