July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Quantitative method for testing toxicity of ocular endo-tamponades in human retina ex-vivo model.
Author Affiliations & Notes
  • Jana D'Amato Tothova
    Research and Development, Alchilife Srl, Ponte San Nicolò, Italy
  • Barbara Ferrari
    Fondazione Banca degli Occhi del Veneto Onlus (FBOV), Italy
  • Laura Giurgola
    Research and Development, Alchilife Srl, Ponte San Nicolò, Italy
  • Claudio Gatto
    Research and Development, Alchilife Srl, Ponte San Nicolò, Italy
  • Footnotes
    Commercial Relationships   Jana D'Amato Tothova, None; Barbara Ferrari, None; Laura Giurgola, None; Claudio Gatto, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 6613. doi:
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      Jana D'Amato Tothova, Barbara Ferrari, Laura Giurgola, Claudio Gatto; Quantitative method for testing toxicity of ocular endo-tamponades in human retina ex-vivo model.. Invest. Ophthalmol. Vis. Sci. 2019;60(9):6613.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : To develop a quantitative method to determine the toxicity of ocular endo-tamponades using human retina ex-vivo model based on MTT viability determination according to ISO 10993-5. The study aimed at determination of retina viability time-course, sample size, positive and negative controls, and the method accuracy and repeatability.

Methods : Human eye globes were recovered and transported to the Eye Bank, Fondazione Banca degli Occhi del Veneto Onlus. The eye globe for research use, was stored in DMEM/F-12 medium at 4°C for 24, 48 and 72 hours before retina extraction. The punch with 2- and 3-mm diameters were evaluated for retina sampling. The tissue viability was determined using TOX-1 In Vitro Toxicology Assay Kit, MTT based (Sigma-Aldrich, Italy) immediately (T0) and 24 h, 48 h, 72 h and 7 days after retina extraction. Sodium Dodecyl Sulfate (SDS) concentrations of 0.25 mg/ml and 0.50 mg/ml were tested as positive (cytotoxic) controls. DMEM/F12 medium was used as negative control.

Results : Retina extracted from eye globe 24 and 48 hours within donor death showed optimal viability at T0 and the mean viability increased up to 178% after 7 days of tissue culture.
Incubation of the samples with 0.25 and 0.50 mg/ml SDS (cytotoxic control) induced 89 ± 4 % and 100 ± 0 % reduction in cell viability at T0, respectively. Samples incubated with DMEM/F-12 showed 2 ± 1 % viability reduction in average at T0. At least eight replicates of 3-mm diameter samples in each group were required in order to obtain the coefficient of variation ≦ 20% within each group of samples. The samples with 2-mm diameter showed significantly higher variability in viability within the group than 3-mm samples.

Conclusions : Quantitative method to assess toxicity of ocular endo-tamponades in vitro, based on the human retina ex-vivo model and the cell viability assay, was standardized with high sensibility a repeatability according to ISO 10993-5.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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