PubMed · 4893560
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F M Polack. 1969. Double corneal forceps.. https://doi.org/10.1016/0002-9394(69)94949-6
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BACKGROUND: During organ culture, corneas swell and have to be de-swollen prior to transplantation using a medium. Both swelling and de-swelling lead to increased endothelial cell loss. Furthermore, dextran, the standard de-swelling substance, is toxic and deposited in the cornea. Whether HES is suitable as a continuous supplement for organ culture and keeps the corneas thin was investigated in this study. METHODS: Forty-five human corneas were stored in MEM plus 7.5% HES 130 for 7, 15, 21, and 28 days. Endothelial cell count was determined before and after organ culture. After storage the water content and amount of adenosine phosphates were measured. RESULTS: Water content was 82.29% after 7 days, 82.98% after 15 days, 81.15% after 21 days, and 83.21% after 28 days. The endothelial cell count decreased by 1.20% after 7 days, 0.60% after 15 days, 4.29% after 21 days, and 6.89% after 28 days. ATP was 0.159 micromol/g dry weight after 28 days of organ culture. CONCLUSION: The water content of corneas stored in HES-containing medium remained constant even after 21 days of organ culture. Endothelial cell loss was less than in comparable studies using a standard medium containing 10% FCS; however, ATP concentrations were worse. HES 130 keeps the corneas thin and can be used as a continuous supplement. It facilitates the handling of corneas during organ culture, making dehydration before transplantation unnecessary, and thus also reduces potential stress factors for the cornea.
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This multicentre study aimed at evaluating the reliability (reproducibility) and relevance (predictivity) of a new commercially available human corneal epithelial (HCE) model (SkinEthic Laboratories, Nice, France) to assess acute ocular irritation. A prevalidation approach (protocol optimisation, transfer and performance) was followed and at each of the four participating laboratories, 20 coded reference chemicals, covering the whole range of irritancy, were tested. The compounds were applied topically to the HCE cultures and the level of cytotoxicity (tissue viability and histological analysis) was determined. Once a standardised protocol was established, a high level of reproducibility between the laboratories was observed. In order to assess the capability of the HCE model to discriminate between irritants (I) and non-irritants (NI), a classification prediction model (PM) was defined based on a viability cut-off value of 60%. The obtained in vitro classifications were compared with different in vivo classifications (e.g. Globally Harmonised System) which were calculated from individual rabbit data described in the ECETOC data bank. Although an overall concordance of 80% was obtained (sensitivity = 100% and specificity = 56%), the predictivity of the HCE model substantially increased when other sources of in vivo and in vitro data were taken into account.