377 Test in Vitro to Investigate the Cytotoxicity and Cellular Nonspecific Stimulation of Basophils with Indomethacin

BACKGROUND: The prevalence of hypersensitivity to nonsteroidal anti-inflammatory drugs (NSAIDs) is high and its in vitro diagnostic is a challenge. The basophil activation test with determination by flow cytometry (FC), of the expression rate of CD63 molecules has been much studied today. NSAIDs several have been evaluated by this technique, which still didn't happen with indomethacin; however, that we may study it is necessary to assess their effects, concentration dependants, on cell viability. We studied the viability of indomethacin dissolved in propylene glycol, analyzing the nonspecific stimulation and cytotoxicity, using in this case the basophil activation test with the use of FC. METHODS: First it was studied the safe concentration of propylene glycol for dilution of indomethacin, incubating basophils from atopic donor with this diluent. In the second phase, the indomethacin was diluted in the following concentrations: 10 mcg/mL, 1 mcg/mL, 0.1 mcg/mL and the CD63 intensity molecules expression was analysed by FC. RESULTS: Regarding the toxicity of propylene glycol, concentrations less than or equal to 0.5% are safe. For indomethacin, the used concentrations (10 mcg/mL, 1 mcg/mL e 0.1 mcg/mL) were viable showing absence of cytotoxicity or nonspecific stimulation. CONCLUSIONS: Propylene glycol as a diluent of indomethacin is necessary to make at concentrations less than or equal to 0.5%. The indomethacin at concentrations of 10 mcg/mL, 1 mcg/mL and 0.1 mcg/mL proved to be not cytotoxic and without nonspecific stimulant action to basophils.