Curran Judith M, Chen Rui, Hunt John A
UK Centre for Tissue Engineering, Clinical Engineering, University of Liverpool, Duncan Building, Daulby Street, Liverpool L69 3GA, UK.
Biomaterials. 2006 Sep;27(27):4783-93. doi: 10.1016/j.biomaterials.2006.05.001. Epub 2006 Jun 2.
Material-driven control of bone-marrow-derived mesenchymal stem cell (MSC) behaviour and differentiation is a very exciting possibility. The aim of this study was to use silane-modified surfaces to control MSC adhesion and differentiation in vitro and evaluate the use of such techniques to control MSC behaviour both in basal and stimulated conditions. A range of characterised clean glass silane-modified surfaces, methyl (-CH(3)), amino (-NH(2)), silane (-SH), hydroxyl (-OH) and carboxyl (-COOH), were produced and cultured in contact with human MSC, in conjunction with a clean glass (TAAB) control, for time periods up to 28 days in basal, chondrogenic and osteogenic stimulated media. The samples were analysed for levels of viable cell adhesion, morphology and the production of various differentiation and transcription markers using both fluorescent immunohistochemistry (collagen I, II, osteocalcin, CBFA1) and real-time polymerase chain reaction (PCR) (collagen I, II, osteocalcin, osteopontin, osteonectin, CBFA1 and Sox 9). Analysis of the results demonstrated that the range of materials could be broken down into three distinct categories. Firstly, the -TAAB control and -CH(3) surfaces maintained the MSC phenotype; secondly, the -NH(2) and -SH-modified surfaces promoted and maintained osteogenesis both in the presence and absence of biological stimuli. These surfaces did not support long-term chondrogenesis under any test conditions. Finally, the -OH and -COOH-modified surfaces promoted and maintained chondrogenesis under both basal and chondrogenic stimulated conditions, but did not support osteogenesis. These results demonstrate that intricate material properties such as surface chemistry and energy can influence MSC behaviour in vitro. These results have implications not only in promoting the efficiency of tissue-engineered constructs, but also to the wider field of MSC isolation, maintenance and expansion.
通过材料驱动来控制骨髓间充质干细胞（MSC）的行为和分化是一种非常令人兴奋的可能性。本研究的目的是使用硅烷修饰的表面在体外控制MSC的黏附与分化，并评估此类技术在基础条件和刺激条件下对MSC行为的控制作用。制备了一系列具有特征的清洁玻璃硅烷修饰表面，包括甲基（-CH(3)）、氨基（-NH(2)）、硅烷（-SH）、羟基（-OH）和羧基（-COOH），并将其与人类MSC接触培养，同时设置清洁玻璃（TAAB）对照，在基础、软骨生成和成骨刺激培养基中培养长达28天。使用荧光免疫组织化学（胶原蛋白I、II、骨钙素、CBFA1）和实时聚合酶链反应（PCR）（胶原蛋白I、II、骨钙素、骨桥蛋白、骨连接蛋白、CBFA1和Sox 9）分析样品的活细胞黏附水平、形态以及各种分化和转录标志物的产生情况。结果分析表明，这些材料可分为三个不同类别。首先，TAAB对照和-CH(3)表面维持了MSC的表型；其次，-NH(2)和-SH修饰的表面在有或无生物刺激的情况下均促进并维持了成骨作用。在任何测试条件下，这些表面都不支持长期软骨生成。最后，-OH和-COOH修饰的表面在基础和软骨生成刺激条件下均促进并维持了软骨生成，但不支持成骨作用。这些结果表明复杂的材料特性，如表面化学和能量，可以在体外影响MSC的行为。这些结果不仅对提高组织工程构建体的效率有意义，而且对MSC分离、维持和扩增的更广泛领域也有意义。
