Department of Biomedical Engineering, Tufts University, Medford, Massachusetts 02155, USA.
J Biomed Mater Res B Appl Biomater. 2011 Oct;99(1):89-101. doi: 10.1002/jbm.b.31875. Epub 2011 Jun 21.
Silk fibroin has been successfully used as a biomaterial for tissue regeneration. To prepare silk fibroin biomaterials for human implantation a series of processing steps are required to purify the protein. Degumming to remove inflammatory sericin is a crucial step related to biocompatibility and variability in the material. Detailed characterization of silk fibroin degumming is reported. The degumming conditions significantly affected cell viability on the silk fibroin material and the ability to form three-dimensional porous scaffolds from the silk fibroin, but did not affect macrophage activation or β-sheet content in the materials formed. Methods are also provided to determine the content of residual sericin in silk fibroin solutions and to assess changes in silk fibroin molecular weight. Amino acid composition analysis was used to detect sericin residuals in silk solutions with a detection limit between 1.0 and 10% wt/wt, while fluorescence spectroscopy was used to reproducibly distinguish between silk samples with different molecular weights. Both methods are simple and require minimal sample volume, providing useful quality control tools for silk fibroin preparation processes.
丝素蛋白已成功用作组织再生的生物材料。为了将丝素蛋白生物材料用于人体植入，需要进行一系列的处理步骤来纯化蛋白质。脱胶以去除炎症性丝胶是与生物相容性和材料变异性相关的关键步骤。本文详细报道了丝素蛋白脱胶的情况。脱胶条件显著影响丝素蛋白材料上细胞的存活率和从丝素蛋白形成三维多孔支架的能力，但不影响材料中巨噬细胞的激活或β-折叠含量。还提供了确定丝素蛋白溶液中残留丝胶含量的方法，并评估了丝素蛋白分子量的变化。氨基酸组成分析用于检测丝胶在丝溶液中的残留，检测限在 1.0 到 10%wt/wt 之间，而荧光光谱用于可重复地区分具有不同分子量的丝样品。这两种方法都简单且需要最小的样品体积，为丝素蛋白制备过程提供了有用的质量控制工具。
