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A. Staropoli, C. Le Moal, C. Baudouin, J.–.M. Warnet, P. Rat; Apoptosis and Oxidative Stress Induced by Medical Devices in Ophthalmology: An in vitro Study for Risk Assessment . Invest. Ophthalmol. Vis. Sci. 2005;46(13):744.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Previously we described the interest of DNA probes to assess the biocompatibility–cytotoxicity of IOL (ARVO; 2003). This in vitro study presented sensitive markers as oxidative stress (OS), mitochondrial and lysosomal activities in order to discriminate cellular responses induced by IOL biomaterials. Our purpose was to improve the biocompatibility ISO norm assessment to understand and prevent postoperative complications of cataract surgery. Methods: These tests were performed on living adherent human epithelial lens cells SRA 01/04 cultured on microplates. Cells were incubated with IOL extracts according to the ISO 10993–12 norm. The IOL biomaterials tested were PMMA, hydrophilic and hydrophobic acrylics. Reference materials were High Density PolyEthylene as negative control, and latex as positive control (ISO 10993–12). Reactive oxygen species and superoxide anion were respectively evaluated with H2DCF–DA and hydroethydine. Membrane integrity, apoptosis and proliferation, mitochondrial and lysosomal activities were respectively assessed with neutral red, Hoechst 33342 and Propidium Iodide, NAO and pNPP quantified by cytofluorometry (Safire, Tecan®). Morphology analyses were performed on confocal microscopy (phalloidin, propidium iodide). Results: We confirmed the inertia of PEHD. Latex eluate induced apoptosis with a decrease of cellular viability (>50%) and an increase of DNA signal due to chromatin condensation confirmed by microscopy. There was no significant variation of cellular viability and DNA after incubation with biomaterials eluates. However, a statistically significant increase in ROS production revealed OS in cells incubated with the eluates of Acrysof (42%) and PMMA (25%) not due to O2.–. Variations of mitochondrial and lysosomal activity were observed vs control. Conclusions: OS, lysosomal and mitochondrial activity are sensitive markers to discriminate biomaterials. This multiparametric study based on international recommendations (ICCVAM) confirmed the limit of the current reviewed ISO 10993 norm and the interest of ROS and metabolism modifications assessment to evaluate biocompatibility–cytotoxicity of medical devices.
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