Department of Health Sciences, University of Piemonte Orientale A. Avogadro, Novara, Italy.
J Biomed Mater Res A. 2012 Sep;100(9):2373-81. doi: 10.1002/jbm.a.34180. Epub 2012 Apr 24.
The interactions between the surface of synthetic scaffolds and cells play an important role in tissue engineering applications. To improve these interactions, two strategies are generally followed: surface coating with large proteins and surface grafting with small peptides. The proteins and peptides more often used and derived from the extracellular matrix, are fibronectin, laminin, and their active peptides, RGD and SIKVAV, respectively. The aim of this work was to compare the effects of coating and grafting of poly(L-lactide) (PLLA) films on MRC5 fibroblast cells. Grafting reactions were verified by X-ray photoelectron spectroscopy. Cell adhesion and proliferation on coated and grafted PLLA surfaces were measured by cell counting. Vinculin localization and distribution were performed on cell cultured on PLLA samples using a fluorescence microscopy technique. Finally, western blot was performed to compare signals of cell adhesion proteins, such as vinculin, Rac1, and RhoA, as well as cell proliferation, such as PCNA. These tests showed similar results for fibronectin and laminin coated PLLA, while RGD grafting is more effective compared with SIKVAV grafting. Considering the overall view of these results, although coating and grafting can both be regarded as effective methods for surface modification to enhance cell adhesion and proliferation on a biomaterial, RGD grafted PLLA show better cell adhesion and proliferation than coated PLLA, while SIKVAV grafted PLLA show similar adhesion but worse proliferation. These data verified different biological effects depending on the surface modification method used.
合成支架表面与细胞的相互作用在组织工程应用中起着重要作用。为了改善这些相互作用，通常采用两种策略：用大蛋白进行表面涂层和用小肽进行表面接枝。更常使用和衍生自细胞外基质的蛋白质和肽分别是纤连蛋白、层粘连蛋白及其活性肽 RGD 和 SIKVAV。本工作旨在比较涂层和接枝聚（L-乳酸）（PLLA）薄膜对 MRC5 成纤维细胞的影响。通过 X 射线光电子能谱验证接枝反应。通过细胞计数测量涂层和接枝 PLLA 表面上的细胞粘附和增殖。使用荧光显微镜技术在 PLLA 样品上培养细胞，检测黏着斑蛋白（如黏着斑蛋白、Rac1 和 RhoA）和细胞增殖（如 PCNA）的定位和分布。最后，通过 Western blot 比较细胞黏附蛋白（如黏着斑蛋白、Rac1 和 RhoA）和细胞增殖（如 PCNA）的信号。这些测试表明，纤连蛋白和层粘连蛋白涂层 PLLA 的结果相似，而 RGD 接枝比 SIKVAV 接枝更有效。考虑到这些结果的整体情况，尽管涂层和接枝都可以被视为增强生物材料上细胞粘附和增殖的有效表面改性方法，但与涂层 PLLA 相比，RGD 接枝 PLLA 显示出更好的细胞粘附和增殖能力，而 SIKVAV 接枝 PLLA 显示出相似的粘附但更差的增殖。这些数据验证了根据所使用的表面改性方法的不同，会产生不同的生物学效应。
