Chirila Traian V, Suzuki Shuko, Bray Laura J, Barnett Nigel L, Harkin Damien G
Queensland Eye Institute, South Brisbane, Queensland, 4101, Australia.
Faculty of Science and Engineering, Queensland University of Technology, Brisbane, Queensland, 4001, Australia.
Prog Biomater. 2013 Nov 28;2(1):14. doi: 10.1186/2194-0517-2-14.
Sericin and fibroin are the two major proteins in the silk fibre produced by the domesticated silkworm, Bombyx mori. Fibroin has been extensively investigated as a biomaterial. We have previously shown that fibroin can function successfully as a substratum for growing cells of the eye. Sericin has been so far neglected as a biomaterial because of suspected allergenic activity. However, this misconception has now been dispelled, and sericin's biocompatibility is currently indisputable. Aiming at promoting sericin as a possible substratum for the growth of corneal cells in order to make tissue-engineered constructs for the restoration of the ocular surface, in this study we investigated the attachment and growth in vitro of human corneal limbal epithelial cells (HLECs) on sericin-based membranes. Sericin was isolated and regenerated from the silkworm cocoons by an aqueous procedure, manufactured into membranes, and characterized (mechanical properties, structural analysis, contact angles). Primary cell cultures from two donors were established in serum-supplemented media in the presence of murine feeder cells. Membranes made of sericin and fibroin-sericin blends were assessed in vitro as substrata for HLECs in a serum-free medium, in a cell attachment assay and in a 3-day cell growth experiment. While the mechanical characteristics of sericin were found to be inferior to those of fibroin, its ability to enhance the attachment of HLECs was significantly superior to fibroin, as revealed by the PicoGreen assay. Evidence was also obtained that cells can grow and differentiate on these substrata.
丝胶蛋白和丝素蛋白是家蚕（Bombyx mori）所产丝纤维中的两种主要蛋白质。丝素蛋白作为一种生物材料已得到广泛研究。我们之前已经表明，丝素蛋白能够成功地作为眼部细胞生长的基质。由于存在潜在的致敏活性，丝胶蛋白作为生物材料至今一直被忽视。然而，这种误解现在已经消除，丝胶蛋白的生物相容性目前是无可争议的。为了促进丝胶蛋白作为角膜细胞生长的可能基质，以便制造用于修复眼表的组织工程构建体，在本研究中，我们研究了人角膜缘上皮细胞（HLECs）在基于丝胶蛋白的膜上的体外附着和生长情况。通过水相法从蚕茧中分离并再生丝胶蛋白，制成膜，并对其进行表征（力学性能、结构分析、接触角）。在存在小鼠饲养细胞的情况下，在补充血清的培养基中建立来自两名供体的原代细胞培养物。在无血清培养基中，在细胞附着试验和为期3天的细胞生长实验中，评估由丝胶蛋白和丝素蛋白 - 丝胶蛋白共混物制成的膜作为HLECs的基质。虽然发现丝胶蛋白的力学特性不如丝素蛋白，但其增强HLECs附着的能力明显优于丝素蛋白，如PicoGreen测定所显示。还获得了细胞可以在这些基质上生长和分化的证据。
