Modified aminosilane substrates to evaluate osteoblast attachment, growth, and gene expression in vitro.
Modified aminosilane substrates to evaluate osteoblast attachment, growth, and gene expression in vitro.
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修饰氨基硅烷底物，用于评估体外成骨细胞附着、生长和基因表达。
DOI：
10.1002/jbm.a.30731
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Landis,WJ
中科院分区：
文献类型：
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作者：
Siperko,LM;Jacquet,R;Landis,WJ
骨细胞-基质的相互作用对于理解骨植入物的设计、选择和表面修饰非常重要。为了深入了解这种相互作用，设计了表面物种近似骨基质生理环境的基质。在三个这样的表面上生长的成骨细胞（Ob）被用来评估与对照组相比，细胞-基质对附着、生长和基因表达的影响。最初的表面制备包括用氨基丙基三乙氧基硅烷（APTES）涂覆玻片，之后用从正常矿化的禽腱中获得的富含胶原蛋白的细胞外基质成分进行修饰：精氨酸-甘氨酸-天冬氨酸（arg-gly-asp）三肽，或由含有通常在血液（模拟体液）中发现的无机离子的亚稳溶液形成的沉淀。每种改性底物，以及未改性（APTES）对照，为从14天龄正常胚鸡获得的初代成骨细胞提供了明显不同的物理（rms粗糙度的差异）和化学表面。在21天的时间内，根据Ob生长和生长速率、碱性磷酸酶（ALP）活性以及I型胶原（COL I）、骨桥蛋白（OPN）、骨钙素（OC）和骨唾液蛋白（BSP）的基因表达，评估细胞对每种底物的反应。这些初步实验表明，本研究中的细胞附着和生长可能是独立的过程，这一假设迫使我们需要进一步的研究。当考虑到细胞生长速度、ALP活性和基因表达时，富含胶原的基质修饰底物具有明显的优势；与APTES对照或其他修饰底物上的细胞相比，这些底物上的细胞ALP活性增加，BSP、OPN和OC的表达增强。这些结果表明，本研究中使用的基质修饰基质为组织生成提供了有利的模板，这表明它们在骨植入物表面设计中的潜力。©2006 Wiley期刊公司[J]生物医学工程学报，2006
Bone cell‐substrate interactions are important to understand in the design, selection, and surface modification of bone implants. To gain insight into such interactions, substrates designed with surface species approximating the physiological environment of bone matrix were studied. Osteoblasts (Ob) grown on three such surfaces were used to evaluate cell‐substrate effects on attachment, growth, and gene expression as compared with controls. Initial surface preparation consisted of coating glass slides with aminopropyltriethoxy silane (APTES), after which the coated slides were modified with collagen‐rich extracellular matrix components obtained from normally mineralizing avian tendon: the tripeptide arginine–glycine–aspartic acid (arg–gly–asp), or a precipitate formed from a metastable solution containing inorganic ions normally found in blood (simulated body fluid). Each of the modified substrates, as well as the nonmodified (APTES) control, provided distinctly different physical (evidenced by differences in rms roughness) and chemical surfaces for seeding primary osteoblasts obtained from 14‐day‐old normal embryonic chickens. Cell responses to each of the substrates were evaluated over a 21‐day period in terms of Ob growth and growth rate, alkaline phosphatase (ALP) activity, and gene expression of type I collagen (COL I), osteopontin (OPN), osteocalcin (OC), and bone sialoprotein (BSP). From these preliminary experiments, indications are that cell attachment and growth in this study possibly are independent processes, an assumption that compels the need for further studies. Collagen‐rich matrix‐modified substrates had a distinct advantage over others when cell growth rate, ALP activity, and gene expression were considered; cells on these substrates exhibited increased ALP activity and enhanced expression of BSP, OPN, and OC when compared with those of cells on APTES controls or other modified substrates. These results indicate that matrix‐modified substrates such as those used in this study provide favorable templates for tissue generation, suggesting their potential in the design of surfaces for bone implants. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res, 2006