Kaufmann E A, Ducheyne P, Shapiro I M
Department of Bioengineering, Center for Bioactive Materials and Tissue Engineering, University of Pennsylvania, 3320 Smith Walk, Philadelphia, Pennsylvania 19104, USA.
J Biomed Mater Res. 2000 Dec 15;52(4):783-96. doi: 10.1002/1097-4636(20001215)52:4<783::aid-jbm24>3.0.co;2-j.
The objective of this study was to determine the effect of porous bioactive glass (45S5) substrate characteristics on the expression and maintenance of the osteoblastic phenotype. We cultured ROS 17/2. 8 cells on substrates with different pore size and porosity for periods up to 14 days and analyzed the characteristics of the cells and extracellular matrix. Results of the study show that the glass substrates supported the proliferation and growth of osteoblast-like cells. Although the morphologies of the cells differed on the various substrates, their shape and the extent of membrane ruffling suggested that they maintained high levels of metabolic activity. Cells on all substrates expressed high levels of alkaline phosphatase activity and produced extracellular matrices that mineralized to form nonstoichiometric, carbonated, calcium-deficient apatites. An important finding was that at a given porosity of 44%, the pore size neither directed nor modulated the in vitro expression of the osteoblastic phenotype. In contrast, porosity did affect cellular function. We noted that at an average pore size of 92 microm, as the porosity increased from 35 to 59%, osteoblast activity was reduced. As designed in this experiment, an increase in the porosity led to a corresponding increase in total surface area of the specimens. With increasing porosity and surface area, glass reactions in the media may persist for longer durations at higher intensities, thereby affecting local media composition. As such, we suggest that extensive conditioning treatments before cell seeding can reduce this effect. Our results also revealed that the expression of the osteoblastic phenotype is enhanced by the ongoing glass dissolution. The reaction pathway at the origin of this effect still needs to be elucidated. Taken together, the findings support the overall hypothesis that in vitro cell activity can be controlled by a careful selection of substrate properties.
本研究的目的是确定多孔生物活性玻璃(45S5)基质特性对成骨细胞表型表达和维持的影响。我们将ROS 17/2.8细胞在具有不同孔径和孔隙率的基质上培养长达14天,并分析细胞和细胞外基质的特性。研究结果表明,玻璃基质支持成骨样细胞的增殖和生长。尽管细胞在不同基质上的形态不同,但其形状和膜褶皱程度表明它们保持着高水平的代谢活性。所有基质上的细胞均表达高水平的碱性磷酸酶活性,并产生矿化形成非化学计量的、含碳酸根的、缺钙磷灰石的细胞外基质。一个重要的发现是,在给定孔隙率为44%时,孔径既不引导也不调节成骨细胞表型的体外表达。相反,孔隙率确实影响细胞功能。我们注意到,在平均孔径为92微米时,随着孔隙率从35%增加到59%,成骨细胞活性降低。如本实验所设计,孔隙率的增加导致标本总表面积相应增加。随着孔隙率和表面积的增加,培养基中的玻璃反应可能会以更高的强度持续更长时间,从而影响局部培养基成分。因此,我们建议在细胞接种前进行广泛预处理可以减少这种影响。我们的结果还表明,持续的玻璃溶解可增强成骨细胞表型的表达。这种效应起源的反应途径仍有待阐明。综上所述,这些发现支持了这样一个总体假设,即体外细胞活性可以通过仔细选择基质特性来控制。