Effects of multigrooved surfaces on fibroblast behavior.
Effects of multigrooved surfaces on fibroblast behavior.
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DOI：
10.1002/jbm.a.10521
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发表时间：
2003-06
期刊：
影响因子：
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通讯作者：
M. Yoshinari;K. Matsuzaka;T. Inoue;Y. Oda;M. Shimono
中科院分区：
文献类型：
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作者：
M. Yoshinari;K. Matsuzaka;T. Inoue;Y. Oda;M. Shimono
微沟槽已被研究作为控制细胞排列的底物。然而，对于控制细胞外基质（ECM）（如胶原）的取向，它们相对太窄和太浅。多沟槽是微沟槽和大沟槽的结合，有望同时控制细胞和ECM的方向。本研究研究了一种制造多沟槽的方法，并评估了成纤维细胞在这些新表面上的行为。在金合金金属模具上制作多凹槽图案，在梯形凹槽上切割90度2微米间距的v型微凹槽。大凹槽的脊宽为50微米，壁宽为50微米，底宽为50微米，深度为25微米。这些凹槽是由一台使用单晶金刚石的超精密微型机器制造的。这个金属模具用作聚苯乙烯表面复制品的模板。微沟槽和光滑的聚苯乙烯复制品也被制备为比较底物。将小鼠成纤维细胞L929细胞在每种复制基质中培养7 ~ 21天。这些时间后，用2.5%戊二醛固定细胞，用常规方法处理，最后用扫描电镜观察。用共聚焦激光扫描显微镜观察ECM的形成。多沟槽金属模具显示出所需的尖锐结构，没有缺陷。在聚苯乙烯复制品上的多沟槽尺寸值与设计值基本一致。在多沟槽和微沟槽基质上培养7天后，成纤维细胞与表面沟槽平行排列。与微沟槽和平面相比，经过21天的培养，沿着多沟槽产生了致密的细胞外基质。这些结果表明，多沟槽可以控制ECM和细胞的取向，从而提高ECM的产生。
Microgrooves have been investigated as substrates for the control of cell alignment. However, they are relatively too narrow and shallow for controlling the orientation of extracellular matrices (ECM) such as collagen. Multigrooves, a combination of microgrooves and macrogrooves, are expected to be able to control the orientation of both cells and ECM. This study investigated a method for fabricating multigrooves and evaluated fibroblast behavior on these novel surfaces. Multigrooved patterns were fabricated on a gold-alloy metal die, in which 90-degree V-shaped microgrooves with a 2-microm pitch were cut on trapezoidal macrogrooves. The macrogrooves had a 50- microm ridge width, a 50-microm wall width, a 50-microm bottom width, and a 25-microm depth. The grooves were made by an ultraprecision micromachine using a single crystal diamond. This metal die served as a template for making surface replicas from polystyrene. Microgrooved and smooth polystyrene replicas also were prepared as comparative substrates. Mouse fibroblast L929 cells were cultured in each type of replica substrate for 7 to 21 days. After these periods, the cells were fixed with 2.5% glutaraldehyde, treated with conventional methods, and, finally, observed by SEM. Confocal laser scanning microscopy was performed to investigate ECM formation. The multigrooved metal die exhibited the desired sharp configuration without defects. The dimensional values of the multigrooves on the polystyrene replicas were almost the same as the designed values. The fibroblasts on the multigrooved and microgrooved substrates were aligned parallel to the surface grooves after 7 days of incubation. In contrast to the microgrooved and flat surfaces, a dense extracellular matrix was produced along the multigrooves after 21 days of incubation. These results suggest that multigrooves can control the orientation of ECM as well as cells and thus enhance the production of ECM.