Author/Authors :
Chirila, Traian V University of Western Australia - Faculty of Science, Australia , Chirila, Traian V Queensland Eye Institute, Australia , Chirila, Traian V Queensland University of Technology - Faculty of Science and Engineering, Australia , Chirila, Traian V University of Queensland - Faculty of Health Sciences, Australia , Suzuki, Shuko Queensland Eye Institute, Australia , Bray, Laura J Leibniz Institute for Polymer Research - Max Bergmann Center of Biomaterials, Germany , Bray, Laura J Queensland Eye Institute, Australia , Bray, Laura J Queensland University of Technology - Faculty of Health, Australia , Barnett, Nigel L University of Queensland - UQ Centre for Clinical Research, Australia , Barnett, Nigel L Queensland Eye Institute, Australia , Barnett, Nigel L Queensland University of Technology - Faculty of Health, Australia , Harkin, Damien G Queensland University of Technology - Faculty of Health, Australia , Harkin, Damien G Institute of Health and Biomedical Innovation, Australia , Harkin, Damien G Queensland Eye Institute, Australia
Abstract :
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.
Keywords :
Silk , Silk proteins , Sericin , Corneal limbal epithelial cells , Cell attachment
