Li Lili, Li Xia, Chen Lu, Sun Peng, Hao Na, Jiang Bo
a National Engineering Research Centre for Biomaterials , Sichuan University , Chengdu , P. R. China.
b Key Laboratory for Radiation Physics and Technology of the Ministry of Education, Institute of Nuclear Science and Technology , Sichuan University , Chengdu , P. R. China.
J Biomater Sci Polym Ed. 2016;27(7):581-98. doi: 10.1080/09205063.2015.1136919. Epub 2016 Feb 21.
The topographic cues of the extracellular matrix may have significant effects upon cellular behavior, such as adhesion, spreading, migration, proliferation, differentiation, and in particular, morphology and orientation. In this study, we examined the effects of microgrooved collagen membrane (MCM) on mesenchymal stem cell (MSC) behavior. The MCM (9 μm in periodicity, and 1-2 μm in depth) was fabricated on an untreated (nonpolar) and smooth polystyrene substrate, based on the absorption and self-assembly properties of collagen on the polystyrene substrate. Methyl thiazolyl tetrazolium assay revealed that cell proliferation on the MCM was enhanced compared with the smooth collagen membrane at day 2. Qualitative observation of MSC behavior using confocal laser scanning microscopy and scanning electron microscopy showed that MSCs grew with a highly elongated morphology and were aligned strictly along the direction of the microgrooves. Additionally, scanning electron microscopy revealed the oriented cells produced a collagenous matrix on the MCM that had a preferential orientation, whereas the collagenous matrix produced by randomly oriented MSCs on the smooth collagen membrane was disorganized. Future studies should investigate the fabrication of oriented topographical substrates, based on the natural biomaterial collagen, to guide cell alignment and oriented growth along definite directions. These substrates may help produce aligned collagenous matrices that could have good potential for the production of tissue substitutes.
细胞外基质的拓扑学线索可能对细胞行为产生重大影响，如黏附、铺展、迁移、增殖、分化，尤其是形态和取向。在本研究中，我们检测了微槽胶原膜（MCM）对间充质干细胞（MSC）行为的影响。基于胶原在聚苯乙烯底物上的吸附和自组装特性，在未经处理（非极性）且光滑的聚苯乙烯底物上制备了MCM（周期为9μm，深度为1 - 2μm）。噻唑蓝比色法显示，在第2天时，与光滑胶原膜相比，MCM上的细胞增殖增强。使用共聚焦激光扫描显微镜和扫描电子显微镜对MSC行为进行的定性观察表明，MSC以高度伸长的形态生长，并严格沿微槽方向排列。此外，扫描电子显微镜显示，定向排列的细胞在MCM上产生了具有优先取向的胶原基质，而随机取向的MSC在光滑胶原膜上产生的胶原基质则是无序的。未来的研究应基于天然生物材料胶原，研究定向拓扑底物的制备，以引导细胞沿确定方向排列和定向生长。这些底物可能有助于产生排列整齐的胶原基质，这在组织替代物的生产中可能具有良好的潜力。
