MacDonald D E, Rapuano B E, Deo N, Stranick M, Somasundaran P, Boskey A L
Langmuir Center for Colloids and Interfaces, School of Engineering, Columbia University, New York, NY 10027, USA.
Biomaterials. 2004 Jul;25(16):3135-46. doi: 10.1016/j.biomaterials.2003.10.029.
The microstructure, chemical composition and wettability of thermally and chemically modified Ti-6Al-4V alloy disks were characterized and correlated with the degree of radiolabeled fibronectin-alloy surface adsorption and subsequent adhesion of osteoblast-like cells. Heating either in pure oxygen or atmosphere (atm) resulted in an enrichment of Al and V within the surface oxide. Heating (oxygen/atm) and peroxide treatment both followed by butanol treatment resulted in a reduction in content of V, but not in Al. Heating (oxygen/atm) or peroxide treatment resulted in a thicker oxide layer and a more hydrophilic surface when compared with passivated controls. Post-treatment with butanol, however, resulted in less hydrophilic surfaces than heating or peroxide treatment alone. The greatest increases in the adsorption of radiolabeled fibronectin following treatment were observed with peroxide/butanol-treated samples followed by peroxide/butanol and heat/butanol, although binding was only increased by 20-40% compared to untreated controls. These experiments with radiolabeled fibronectin indicate that enhanced adsorption of the glycoprotein was more highly correlated with changes in chemical composition, reflected in a reduction in V content and decrease in the V/Al ratio, than with changes in wettability. Despite promoting only a modest elevation in fibronectin adsorption, the treatment of disks with heat or heat/butanol induced a several-fold increase in the attachment of MG63 cells promoted by a nonadhesive concentration of fibronectin that was used to coat the pretreated disks compared to uncoated disks. Therefore, results obtained with these modifications of surface properties indicate that an increase in the absolute content of Al and/or V (heat), and/or in the Al/V ratio (with little change in hydrophilicity; heat+butanol) is correlated with an increase in the fibronectin-promoted adhesion of an osteoblast-like cell line. It would also appear that the thermal treatment-induced enhancement of cell adhesion in the presence of this integrin-binding protein is due to its increased biological activity, rather than a mass effect alone, that appear to be associated with changes in chemical composition of the metallic surface. Future studies will investigate the influence of the surface chemical composition of various implantable alloys on protein adsorption and receptor-mediated cell adhesion. In addition, by altering the properties of bound osteogenic protein enhancing exposure to cell integrin binding domains, it may be possible to develop implant surfaces which enhance the attachment, adhesion and developmental response of osteoblast precursors leading to accelerated osseointegration.
对经热改性和化学改性的Ti-6Al-4V合金圆盘的微观结构、化学成分和润湿性进行了表征,并将其与放射性标记的纤连蛋白在合金表面的吸附程度以及随后成骨样细胞的黏附情况相关联。在纯氧或大气环境中加热会导致表面氧化物中Al和V的富集。先进行加热(氧气/大气环境)和过氧化物处理,随后进行丁醇处理,会使V的含量降低,但Al的含量不变。与钝化对照相比,加热(氧气/大气环境)或过氧化物处理会导致形成更厚的氧化层和更亲水的表面。然而,与单独的加热或过氧化物处理相比,用丁醇进行后处理会使表面亲水性降低。在处理后的样品中,观察到放射性标记的纤连蛋白吸附增加最多的是过氧化物/丁醇处理的样品,其次是过氧化物/丁醇和加热/丁醇处理的样品,不过与未处理的对照相比,结合仅增加了20%-40%。这些使用放射性标记纤连蛋白的实验表明,糖蛋白吸附的增强与化学成分的变化更高度相关,表现为V含量的降低和V/Al比值的减小,而非与润湿性的变化相关。尽管热或热/丁醇处理圆盘仅使纤连蛋白吸附适度增加,但与未包被圆盘相比,用非黏附浓度的纤连蛋白包被预处理圆盘后,MG63细胞的附着增加了数倍。因此,这些表面性质改性的结果表明,Al和/或V的绝对含量增加(加热)和/或Al/V比值增加(亲水性变化不大;加热+丁醇)与纤连蛋白促进的成骨样细胞系黏附增加相关。还似乎在这种整合素结合蛋白存在的情况下,热处理诱导的细胞黏附增强是由于其生物活性增加,而非仅仅是质量效应,这似乎与金属表面化学成分的变化有关。未来的研究将调查各种可植入合金的表面化学成分对蛋白质吸附和受体介导的细胞黏附的影响。此外,通过改变结合的成骨蛋白的性质,增强对细胞整合素结合域的暴露,有可能开发出能够增强成骨细胞前体的附着、黏附和发育反应从而加速骨整合的植入表面。