Takata T, Wang H L, Miyauchi M
Department of Oral Pathology, Hiroshima University Faculty of Dentistry, 1-2-3 Kasumi, Minami-ku, Hiroshima, Japan.
Clin Oral Implants Res. 2001 Aug;12(4):332-8. doi: 10.1034/j.1600-0501.2001.012004332.x.
To evaluate the biological effects of guided bone regeneration (GBR) barrier materials on osteoblastic cell migration, migration of mouse osteoprogenitor cells (MC3T3-E1) was examined, in vitro, on various membranes. Eight commercially available GBR membranes - bovine type I collagen (BioMend; BM), porcine type I collagen (BioGide; BG), bovine type I atelocollagen (Tissue Guide; TG), polylactic acid (Epi-Guide; EG), co-polymer of polylactic acid and polyglycolic acid (Resolute; RL, Resolut XT; RL-XT), expanded polytetrafluoroethylene (e-PTFE; Gore Tex; GT) and co-polymer of cellulose acetate and nitrocellulose (Millipore filter; MP) - were tested. A 3x5 mm section of the membrane was fixed to the bottom of a culture dish with double-sided adhesive tape, and half of the membrane was closely covered by PARAFILM (American National Can) to leave an unexposed area for cell migration. The border between exposed and unexposed areas was marked as a baseline of cell migration. Membranes were then plated with 3 ml of cell suspension at an initial density of 1x105 cells/ml in alpha-MEM culture medium with 10% fetal bovine serum and ascorbic acid. After a 5-hour incubation, non-attached cells were completely washed out with phosphate buffered saline and the PARAFILM cover was removed. After 3 days cultivation, specimens were fixed with 10% buffered formalin and stained briefly with hematoxylin. The area of cell migration on a membrane was analyzed using a LA 500 Image Analysis System and migration area per unit length of the baseline (mm2/mm) was compared among membranes. Results demonstrated that cell migration was greater in the order: RL>RL-XT, BM, TG, MP>EG, BG. Membranes except for BG, EG and GT showed the migration rate equal to or higher than a plastic culture cover slip (Celldesk) (P<0.01) on which cells generally grow favorably. Only a small number of the cells attached to GT, and the net cell migration for the membrane could not be determined. These results indicate that GBR barrier materials per se may influence the process of bone regeneration in vivo through the effects of their presence on cell migration.
为评估引导骨再生(GBR)屏障材料对成骨细胞迁移的生物学效应,在体外检测了小鼠骨祖细胞(MC3T3-E1)在各种膜上的迁移情况。测试了8种市售GBR膜,分别为牛I型胶原(BioMend;BM)、猪I型胶原(BioGide;BG)、牛I型去端胶原(Tissue Guide;TG)、聚乳酸(Epi-Guide;EG)、聚乳酸与聚乙醇酸共聚物(Resolute;RL,Resolut XT;RL-XT)、膨体聚四氟乙烯(e-PTFE;Gore Tex;GT)以及醋酸纤维素与硝酸纤维素共聚物(密理博滤膜;MP)。将3×5 mm的膜片用双面胶带固定于培养皿底部,一半膜片用PARAFILM(美国国家罐头公司)紧密覆盖,留出未暴露区域用于细胞迁移。暴露区域与未暴露区域的边界标记为细胞迁移的基线。然后在膜片上接种3 ml细胞悬液,初始密度为1×105个细胞/ml,培养基为含10%胎牛血清和抗坏血酸的α-MEM培养基。孵育5小时后,用磷酸盐缓冲盐水将未附着细胞彻底洗去,去除PARAFILM覆盖物。培养3天后,用10%中性福尔马林固定标本,并用苏木精短暂染色。使用LA 500图像分析系统分析膜上细胞迁移的面积,并比较各膜基线单位长度的迁移面积(mm2/mm)。结果表明,细胞迁移能力从高到低依次为:RL>RL-XT、BM、TG、MP>EG、BG。除BG、EG和GT外,其他膜的迁移率等于或高于塑料培养盖玻片(Celldesk)(P<0.01),细胞通常在该盖玻片上生长良好。只有少数细胞附着于GT,无法确定该膜的净细胞迁移情况。这些结果表明,GBR屏障材料本身可能通过其存在对细胞迁移的影响,在体内影响骨再生过程。