EFFECT OF TITANIUM SURFACE-ROUGHNESS ON PROLIFERATION, DIFFERENTIATION, AND PROTEIN-SYNTHESIS OF HUMAN OSTEOBLAST-LIKE CELLS (MG63)
EFFECT OF TITANIUM SURFACE-ROUGHNESS ON PROLIFERATION, DIFFERENTIATION, AND PROTEIN-SYNTHESIS OF HUMAN OSTEOBLAST-LIKE CELLS (MG63)
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DOI：
10.1002/jbm.820290314
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发表时间：
1995-03-01
期刊：
影响因子：
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通讯作者：
BOYAN, BD
中科院分区：
文献类型：
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作者：
MARTIN, JY;SCHWARTZ, Z;BOYAN, BD
研究了表面粗糙度对成骨细胞增殖、分化和蛋白质合成的影响。人成骨细胞样细胞（MG 63）培养在钛（Ti）盘，已通过五种不同的治疗方案之一制备。用氢氟酸-硝酸预处理所有盘并洗涤（PT）。PT磁盘还包括：清洗，然后电抛光（EP）;细喷砂，用HCl和H2SO 4蚀刻，然后清洗（FA）;粗喷砂，用HCl和H2SO 4蚀刻，然后清洗（CA）;或Ti等离子喷涂（TPS）。使用标准组织培养塑料作为对照。表面形貌和轮廓进行了评估，明场和暗场显微镜，冷场发射扫描电子显微镜，激光共聚焦显微镜，而化学成分进行了映射使用能量色散X射线分析和元素分布确定使用俄歇电子能谱。通过测定细胞数、[H-3]胸苷掺入DNA、碱性磷酸酶比活性、[H-3]尿苷掺入RNA、[H-3]脯氨酸掺入胶原酶可消化蛋白（CDP）和非胶原酶可消化蛋白（NCP）以及[S-35]硫酸盐掺入蛋白多糖来评价表面粗糙度对细胞的影响。五种不同的钛表面按最光滑到最粗糙的顺序排列：EP、PT、FA、CA和TPS。在所有表面上均发现TiO 2层，其厚度范围从最光滑组的100埃到最粗糙组的300埃。与塑料上的细胞汇合培养物相比，TPS表面上的细胞数量减少，EP表面上的细胞数量增加，而其他表面上的细胞数量与塑料相当。[H-3]胸苷掺入与表面粗糙度呈负相关。碱性磷酸酶比活性在孤立的细胞被发现，随着表面粗糙度的增加，除了那些细胞培养CA。相比之下，细胞层中的酶活性仅在FA和TPS处理的表面上生长的培养物中降低。表面粗糙度与RNA和CDP的产生有直接的相关性，表面粗糙度对NCP的产生没有明显的影响。蛋白聚糖的合成细胞被抑制在所有的表面研究，观察到的CA和EP组中的最大抑制。这些结果表明，表面粗糙度改变成骨细胞的增殖，分化和基质生产在体外。结果还表明，植入物表面粗糙度可能在体内细胞表型表达的决定中发挥作用。(C)John Wiley & Sons，Inc.
The effect of surface roughness on osteoblast proliferation, differentiation, and protein synthesis was examined. Human osteoblast-like cells (MG63) were cultured on titanium (Ti) disks that had been prepared by one of five different treatment regimens. All disks were pretreated with hydrofluoric acid-nitric acid and washed (PT). PT disks were also: washed, and then electropolished (EP); fine sandblasted, etched with HCl and H2SO4, and washed (FA); coarse sandblasted, etched with HCl and H2SO4, and washed (CA); or Ti plasma-sprayed (TPS). Standard tissue culture plastic was used as a control. Surface topography and profile were evaluated by brightfield and darkfield microscopy, cold field emission scanning electron microscopy, and laser confocal microscopy, while chemical composition was mapped using energy dispersion X-ray analysis and elemental distribution determined using Auger electron spectroscopy. The effect of surface roughness on the cells was evaluated by measuring cell number, [H-3]thymidine incorporation into DNA, alkaline phosphatase specific activity, [H-3]uridine incorporation into RNA, [H-3]proline incorporation into collagenase digestible protein (CDP) and noncollagenase-digestible protein (NCP), and [S-35]sulfate incorporation into proteoglycan.Based on surface analysis, the five different Ti surfaces were ranked in order of smoothest to roughest: EP, PT, FA, CA, and TPS. A TiO2 layer was found on all surfaces that ranged in thickness from 100 Angstrom in the smoothest group to 300 Angstrom in the roughest. When compared to confluent cultures of cells on plastic, the number of cells was reduced on the TPS surfaces and increased on the EP surfaces, while the number of cells on the other surfaces was equivalent to plastic. [H-3]Thymidine incorporation was inversely related to surface roughness. Alkaline phosphatase specific activity in isolated cells was found to decrease with increasing surface roughness, except for those cells cultured on CA. In contrast, enzyme activity in the cell layer was only decreased in cultures grown on FA- and TPS-treated surfaces. A direct correlation between surface roughness and RNA and CDP production was found. Surface roughness had no apparent effect on NCP production. Proteoglycan synthesis by the cells was inhibited on all the surfaces studied, with the largest inhibition observed in the CA and EP groups. These results demonstrate that surface roughness alters osteoblast proliferation, differentiation, and matrix production in vitro. The results also suggest that implant surface roughness may play a role in determining phenotypic expression of cells in vivo. (C) 1995 John Wiley & Sons, Inc.