Human corneal endothelial cell growth on a silk fibroin membrane
Authors
Madden PW, Lai JN, George KA, Giovenco T, Harkin DG, Chirila TV
Institution
Queensland Eye Institute, 41 Annerley Road, South Brisbane
Country
Australia
Year
2011
Journal
Biomaterials
Abstract
Tissue engineering of the cornea could overcome shortages of donor corneas for transplantation and improve quality. Our aim was to grow an endothelial layer on a substratum suitable for transplant. Silkworm (Bombyx mori) fibroin was prepared as 5 μm thick transparent membranes. The B4G12 cell line was used to assess attachment and growth of human corneal endothelial cells on fibroin and compare this with a reference substratum of tissue-culture plastic. To see if cell attachment and proliferation could be improved, we assessed coatings of collagen IV, FNC Coating Mix(®) and a chondroitin sulphate-laminin mixture. All the coatings improved the final mean cell count, but consistently higher cell densities were achieved on a tissue-culture plastic rather than fibroin substratum. Collagen-coated substrata were the best of both groups and collagen-coated fibroin was comparable to uncoated tissue-culture plastic. Only fibroin with collagen coating achieved cell confluency. Primary human corneal endothelial cells were then grown using a sphere-forming technique and when seeded onto collagen-coated fibroin they grew to confluency with polygonal morphology. We report the first successful growth of primary human corneal endothelial cells on coated fibroin as a step in evaluating fibroin as a substratum for the transplantation of tissue-constructs for endothelial keratoplasty.
Tissue type
Corneal
Tissue info
Primary human corneal endothelial cells from corneas of deceased donors and the corneoscleral rims