Real-time cellular analysis as a novel approach for in vitro cytotoxicity testing of medical device extracts.

Cytotoxicity measurements are often performed to evaluate the biocompatibility of medical device materials. Here, we describe the use of a real-time cell analyzer (RTCA) system for the investigation of biocompatibility of medical devices by comparing RTCA results to two distinct methods described in the International Organization for Standardization (ISO) guidelines. Mouse L-929 fibroblast proliferation was assessed every 15 min from 24 to 100 h during the pretreatment and postextract addition period. Simultaneously, we performed quantitative cytotoxicity analyses using water-soluble tetrazolium salt (WST-1) and qualitatively scored cytotoxicity by examining changes in morphology at 24-h intervals. The RTCA uses electrical impedance to measure cell viability quantified as a normalized cellular index (CI) which was converted in this study to a reactivity grade. Results from microscopic analyses were expressed as a reactivity grade, based on morphology as defined by the ISO 10993-5:2009. There was a clear correlation between addition of cytotoxic agents and, both, decreased normalized CI and concomitant cell layer destruction observed by microscopy. Results obtained from the colorimetric WST-1 assays also correlated with normalized CI at various time points tested. The results indicate that RTCA allows for automated and accurate assessment of biocompatibility of medical devices and biomaterials.