The Boston Keratoprosthesis: comparing corneal epithelial cell compatibility with titanium and PMMA.
OBJECTIVE
To determine in vitro whether titanium is superior in corneal cell compatibility to standard polymethyl-methacrylate (PMMA) for the Boston Keratoprosthesis (KPro).


METHODS
Human corneal-limbal epithelial (HCLE) cells were cultured 24, 48, 72, 96, 120, 144, or 168 hours in culture plates alone (controls) or with PMMA or titanium discs. Experiments were performed in triplicate and repeated (final n = 6). To determine if a soluble, toxic factor is emitted from materials, concurrent experiments at 48 and 144 hours were performed with discs placed in Transwell Supports, with HCLE cells plated beneath. As an additional test for soluble factors, cells were incubated 24 hours with disc-conditioned media, and number of viable cells per well was quantified at each timepoint by proliferation assay. To determine if delayed cell proliferation was attributable to cell death, HCLE cell death was measured under all conditions and quantified at each timepoint by cytotoxicity assay. The effects of material on HCLE cell proliferation over time was determined by repeated measures ANOVA. P < 0.05 was statistically significant.


RESULTS
HCLE cell proliferation was greater in wells with titanium discs compared to PMMA. Differences between the test discs and control non-disc cocultures were statistically significant over time for both cell proliferation (P = 0.001) and death (P = 0.0025). No significant difference was found using Transwells (P = 0.9836) or disc-conditioned media (P = 0.36).


CONCLUSIONS
This in vitro HCLE cell model demonstrates significantly increased cell proliferation and decreased cell death with cell/titanium contact compared to cell/PMMA contact. Moreover, differences are unlikely attributable to a soluble factor. Prospective in vivo analysis of the two KPro biomaterials is indicated.
Ament
Jared D
JD
Massachusetts Eye and Ear Infirmary; Harvard Medical School, Boston, Massachusetts, USA. Jared_Ament@MEEI.harvard.edu
Spurr-Michaud
Sandra J
SJ
Dohlman
Claes H
CH
Gipson
Ilene K
IK
D001672:Biocompatible Materials; D016923:Cell Death; D049109:Cell Proliferation; D002478:Cells, Cultured; D003315:Cornea; D004847:Epithelial Cells / Q000166:cytology; D019573:Epithelium, Corneal / Q000166:cytology*; D006801:Humans; D016850:Limbus Corneae / Q000166:cytology; D008422:Materials Testing; D019904:Polymethyl Methacrylate; D019736:Prostheses and Implants; D014025:Titanium
D001672:Biocompatible Materials; D019904:Polymethyl Methacrylate; D014025:Titanium