Verma Vipin, Verma Poonam, Ray Pratima, Ray Alok R
Centre for Biomedical Engineering, Indian Institute of Technology Delhi (IITD), New Delhi-110 016, India.
Biomed Mater. 2008 Jun;3(2):025007. doi: 10.1088/1748-6041/3/2/025007. Epub 2008 Apr 15.
Human hair proteins were isolated and purified for the fabrication of tissue-engineering scaffolds. Their cellular compatibility was studied using NIH3T3 mice fibroblast cells. The proteins were characterized using FTIR spectroscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis for molecular weights and two-dimensional polyacrylamide gel electrophoresis for their isoelectric points (pIs). The molecular weights of keratins were in the range of 40-60 kilo-Daltons (kDa) and of matrix proteins were in the range of 15-30 kDa. The pIs of keratins were found to be in the range of 4.5-5.3. Sponges of the proteins were formed by lyophilization. Scanning electron microscopy was performed to examine the surface. Swelling studies were carried out in phosphate buffer saline at physiological pH 7.4. The hydrophilic character of the protein surface was studied by determining an average contact angle, which came to be 37 degrees. The wells of tissue culture plates were coated with these proteins for studying the attachment and morphology of the cells. The protein detachment study was done to ensure the adsorption of proteins on the wells until the completion of the experiments. The cellular growth on a protein-coated surface showed three-dimensional 'bulged' morphology due to cell-cell and cell-matrix contacts. The sponges of human hair proteins supported more cells for a longer period than control. The morphology and cell proliferation studies exhibited by NIH3T3 cells on these proteins have shown their potential to be used as tissue-engineering scaffolds with better cell-cell contacts and leucine-aspartic acid-valine (LDV)-mediated cell-matrix interactions.
为了制造组织工程支架，对人发蛋白进行了分离和纯化。使用NIH3T3小鼠成纤维细胞研究了它们的细胞相容性。利用傅里叶变换红外光谱、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测定分子量以及二维聚丙烯酰胺凝胶电泳测定等电点（pI）对这些蛋白质进行了表征。角蛋白的分子量范围为40-60千道尔顿（kDa），基质蛋白的分子量范围为15-30 kDa。发现角蛋白的pI在4.5-5.3范围内。通过冻干形成蛋白质海绵。进行扫描电子显微镜检查表面。在生理pH值7.4的磷酸盐缓冲盐水中进行溶胀研究。通过测定平均接触角来研究蛋白质表面的亲水性，结果为37度。在组织培养板的孔中涂覆这些蛋白质，以研究细胞的附着和形态。进行蛋白质脱离研究以确保蛋白质在孔上的吸附直至实验完成。由于细胞-细胞和细胞-基质接触，在蛋白质包被表面上的细胞生长呈现三维“凸起”形态。与人发蛋白海绵相比，对照在更长时间内支持更多细胞。NIH3T3细胞在这些蛋白质上表现出的形态和细胞增殖研究表明，它们有潜力用作具有更好细胞-细胞接触和亮氨酸-天冬氨酸-缬氨酸（LDV）介导的细胞-基质相互作用的组织工程支架。
