Division of Engineering, Brown University, Providence, Rhode Island 02917, USA.
J Biomed Mater Res A. 2010 Sep 15;94(4):1042-9. doi: 10.1002/jbm.a.32778.
An in vitro competitive coculture system was designed in this study to investigate vascular cell functions on submicron (lateral feature sizes larger than 100 nm) and nanometer (lateral and vertical feature sizes smaller than 50 nm) rough titanium surface features created via e-beam evaporation with identical chemistry to that of control (unmodified) titanium. Rat aortic endothelial cells (RAEC) and rat aortic smooth muscle cells (RASMC) were individually cultured and cocultured on such substrates for 4 h as well as 1, 3, and 5 days. In addition, RAEC and RASMC intracellular collagen and elastin synthesis were determined for up to 14 days on the various substrates. Compared to control flat titanium surfaces, the results showed for the first time that submicron rough surface features on titanium enhanced RAEC proliferation (followed by nanometer surface features), while simultaneously inhibiting RASMC proliferation. In addition, protein synthesis assays demonstrated enhanced collagen and elastin production by RAEC cultured on submicron rough titanium substrates (followed by nano rough) compared to control flat titanium substrates. Surface characterization results showed that submicron rough featured titanium possessed the greatest hydrophilicity (followed by nanometer surface featured), which may have promoted select protein adsorption to increase RAEC over RASMC function. In contrast to RASMC, RAEC morphology was significantly altered by the underlying titanium surface features, which may also explain the altered vascular cell functions on the different substrates. Thus, this study suggested that submicron to nanometer rough surface features should be further studied for improving titanium for vascular stent applications.
本研究设计了一种体外竞争共培养体系,以研究血管细胞在亚微米(侧向特征尺寸大于 100nm)和纳米(侧向和垂直特征尺寸小于 50nm)粗糙钛表面特征上的功能,这些表面特征是通过电子束蒸发形成的,具有与对照(未修饰)钛相同的化学性质。将大鼠主动脉内皮细胞(RAEC)和大鼠主动脉平滑肌细胞(RASMC)分别在这些底物上进行单独培养和共培养,时间分别为 4 小时、1 天、3 天和 5 天。此外,还在各种底物上测定了 RAEC 和 RASMC 的细胞内胶原和弹性蛋白合成长达 14 天。与对照平面钛表面相比,结果首次表明,钛表面的亚微米粗糙特征增强了 RAEC 的增殖(随后是纳米表面特征),同时抑制了 RASMC 的增殖。此外,蛋白质合成测定表明,与对照平面钛底物相比,在亚微米粗糙钛底物上培养的 RAEC 产生了更多的胶原和弹性蛋白(随后是纳米粗糙)。表面特性研究结果表明,亚微米粗糙特征钛具有最大的亲水性(随后是纳米表面特征),这可能促进了某些蛋白质的吸附,从而增加了 RAEC 相对于 RASMC 的功能。与 RASMC 相反,RAEC 的形态被钛的基底表面特征显著改变,这也可能解释了在不同底物上血管细胞功能的改变。因此,本研究表明,亚微米到纳米粗糙表面特征应该进一步研究,以改善钛在血管支架应用中的性能。