title: Human corneal endothelial cell growth on a silk fibroin membrane.
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.
authors.affiliation: Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101, Australia. peter.madden@qei.org.au
authors.forename: Peter W
authors.initials: PW
authors.lastname: Madden
authors.forename: Jonathan N X
authors.initials: JN
authors.lastname: Lai
authors.forename: Karina A
authors.initials: KA
authors.lastname: George
authors.forename: Talia
authors.initials: T
authors.lastname: Giovenco
authors.forename: Damien G
authors.initials: DG
authors.lastname: Harkin
authors.forename: Traian V
authors.initials: TV
authors.lastname: Chirila
mesh_terms: D000818:Animals; D012831:Bombyx; D002452:Cell Count; D002455:Cell Division; D002460:Cell Line; D003094:Collagen / Q000737:chemistry; D042783:Endothelial Cells / Q000166:cytology*; D004728:Endothelium, Corneal / Q000166:cytology*; D005349:Fibroins / Q000737:chemistry*; D006801:Humans; D007797:Laminin / Q000737:chemistry; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D007797:Laminin; D003094:Collagen; D005349:Fibroins