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Martin Hermel, Sabine Salla, Nicole Hamsley, Peter Walter, Anne Rieck; Comparison of Dehydration Kinetics of Organ Cultured Human Corneas by Different Deswelling Media using Optical Coherence Tomography. Invest. Ophthalmol. Vis. Sci. 2013;54(15):3069.
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© ARVO (1962-2015); The Authors (2016-present)
Human donor corneas experience swelling while in organ culture, and require dehydration by placing in hyperosmolar culture media prior to grafting. In this study we compared the deswelling kinetics of different media by measuring central corneal thickess (CCT) over time using optical coherence tomography (OCT).
The study was conducted in accordance with the Declaration of Helsinki. Ethical approval was obtained. Thirty-three human donor corneas not suitable for transplantation were stored in MEM + antibiotics + 2% fetal calf serum (FCS) for 15 up to 28 days and were then randomly assigned to four groups. Dehydration was induced by using serum-containing deswelling media with dextran (BK2 [“Biochrom K2”, Biochrom, n=8], CJ [“CorneaJet”, gift of EuroBio, n=9] or DM [MEM + antibiotics + 2% FCS + 5% dextran 500, n=8]), or a serum-free medium with an undisclosed deswelling compound (SA3, [“StemAlpha 3”, gift of Stemalpha, n=8]). CCT was measured by OCT (Heidelberg Engineering) before deswelling and after 1, 2, 3, 6, 12, 24, 48, 72 and 144 hours. Data are presented as mean±SD. Dehydration kinetics was analysed via the nonlinear platform in JMP10 Pro.
Prior to deswelling, CCT was 1123±195µm, 1157±74µm, 1133±177µm and 1229±152µm in the BK2, CJ, DM and SA3 groups, respectively. Minimal corneal thickness was obtained after 12h in SA3 (702±207µm), 24h in BK2 (615±99µm) and 48h in CJ (599±37µm) and DM (625±125µm) groups. Final CCT after 144h was 672±114µm, 626±57µm, 666±147µm and 929±149µm in the BK2, CJ, DM and SA3 groups, respectively. The deswelling kinetics was best modelled as a biexponential function (overall r2=0,554) with decay rates of the media differing at the α=0.05 level.
All media displayed dehydration of the corneal stroma and reduction of CCT. SA3 displayed a marked increase in CCT after 12h, possibly due to degradation of the proprietary deswelling agent, and should thus not be used in conjunction with other serum-containing culture media. All dextran containing media obtained minimal CCT values after 24 to 48h deswelling, with a slight subsequent CCT increase in BK2 and DM, which may reflect dextran degradation. Grafting of organ cultured corneas should be performed after 24 to 48h deswelling.
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