Influence of substrate on corneal epithelial cell viability within ocular surface models.
Corneal tissue engineering has improved dramatically over recent years. It is now possible to apply these technological advancements to the development of superior in vitro ocular surface models to reduce animal testing. We aim to show the effect different substrates can have on the viability of expanded corneal epithelial cells and that those which more accurately mimic the stromal surface provide the most protection against toxic assault. Compressed collagen gel as a substrate for the expansion of a human epithelial cell line was compared against two well-known substrates for modelling the ocular surface (polycarbonate membrane and conventional collagen gel). Cells were expanded over 10 days at which point cell stratification, cell number and expression of junctional proteins were assessed by electron microscopy, immunohistochemistry and RT-PCR. The effect of increasing concentrations of sodium lauryl sulphate on epithelial cell viability was quantified by MTT assay. Results showed improvement in terms of stratification, cell number and tight junction expression in human epithelial cells expanded upon either the polycarbonate membrane or compressed collagen gel when compared to a the use of a conventional collagen gel. However, cell viability was significantly higher in cells expanded upon the compressed collagen gel. We conclude that the more naturalistic composition and mechanical properties of compressed collagen gels produces a more robust corneal model.
School of Pharmacy, University of Reading, United Kingdom.
Yun
Y
Feng
James
J
Foster
Shengli
S
Mi
Bo
B
Chen
Che John
CJ
Connon
D002452:Cell Count; D002460:Cell Line; D002470:Cell Survival; D003094:Collagen; D019573:Epithelium, Corneal / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D039663:Integrin beta4 / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D008954:Models, Biological; D010750:Phosphoproteins / Q000378:metabolism; D011076:Polycarboxylate Cement; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D062826:Zonula Occludens-1 Protein
D039663:Integrin beta4; D008565:Membrane Proteins; D010750:Phosphoproteins; D011076:Polycarboxylate Cement; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein; C005506:polycarbonate; D012967:Sodium Dodecyl Sulfate; D003094:Collagen