RESUMEN
Background: Ragweed as an invasive species in Europe has become more important for allergy sufferers in the last decade. Because pollen fractions can be found in the respirable fraction of aerosols, they can generate severe disease progressions. Objective: To obtain information about the concentration and distribution of 1 of the main ragweed allergens Ambrosia artemisiifolia 1 in the air of Vienna, PM10 and PM2.5 fine dust filters were analyzed. Methods: Standard fine dust filters used for air quality monitoring were analyzed via ELISA and immunogold scanning electron microscopy. Results: Via ELISA it was possible to show that already at pollen season start in August a recognizably high A artemisiifolia 1 concentration can be found. In addition, the allergen concentration in the air stays comparatively high after the peak season has ended even when the pollen concentration drops to a moderate level. The immunogold electron microscopy investigation directly applied on filters shows that the allergen can be found on organic as well as on mixtures of organic and inorganic particles. A first semistatistical analysis of the labeled particle sizes indicates that a large number of the allergen carriers can be found within the smallest particle size range. Nevertheless, further investigations are needed to obtain enough particle counts for a significant statistical analysis. Conclusions: It was possible to show that reliable results can be obtained from ELISA and immunogold scanning electron microscopy directly applied on filters that are used in air quality monitoring sites. By adaptation of the used protocols, it should be possible to obtain respective information about further allergens.
RESUMEN
According to standing regulations animal tests are still state of the art for the evaluation of the sensitization potential of medical devices. The aim of our study was to develop an in vitro method that can be used for testing of extracts of medical devices. The novel MDA-ARE assay is a cell based reporter gene assay focused on the ARE-Nrf2 pathway, which is involved in the dermal sensitization process. Optimization of the reporter construct and the cell line resulted in an improvement of the detection limit and a reduction of the incubation time to 6 h, which lowers cytotoxic side effects of the extracts on the cells. Using the assay, 21 out of 22 pure chemicals were identified correctly as skin sensitizers or non-sensitizers. All sensitizers could be detected at far lower concentrations compared to the local lymph node assay, the state-of-the-art animal test. To evaluate the assay's suitability for the testing of medical devices, medical grade silicone containing 0.1% of known skin sensitizers was prepared as positive controls and extracts of these positive controls were tested in comparison to extracts from pure silicone samples. All silicone samples were correctly and reproducibly identified as sensitizing or non-sensitizing demonstrating that the MDA-ARE assay is a sensitive and reliable tool for the detection of skin sensitizers in extracts of medical devices. The developed and validated test protocol was used for medical device extracts and showed its applicability for real samples and thus can contribute to reduce or even to replace the need for animal tests.
