Assessment of test method variables for in vitro skin irritation testing of medical device extracts

Skin irritation is an important component of the biological safety evaluation of medical devices. This testing has typically been performed using in vivo models. However, in an effort to reduce the need for in vivo testing, alternative methods for assessing skin irritation potential in vitro have been developed using a Reconstructed Human Epidermis (RhE) model. During the development of the protocol for the round robin validation of in vitro irritation testing for medical device extracts, it became clear that there were three points in the procedure where different options may be validated within each laboratory for routine testing: sample exposure time (18 vs 24 h), SDS positive control concentration, and cytokine (IL-1 alpha) release testing. The goal of our study was to evaluate the effect of these variables. EpiDerm (TM) tissues were exposed to extracts of three plain polymer samples, and four polymers embedded with known irritant chemicals. Exposures were performed for 18 and 24 h. Resulting tissue viability was assessed by MIT reduction and IL-1 alpha release was assessed by ELISA. Testing was also performed using various concentrations of SDS ranging from 0.5 to 1% (w/v). Overall, results were similar for samples tested and 18 and 24 h, but the 18 h exposure time has the potential to have an impact on the results of some sample types. IL-1 alpha testing was shown to be useful to clarify conflicting tissue viability results. Use of a lower concentration of SDS as a positive control can help prevent issues that arise from excessive tissue damage often caused by 1% SDS.