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Establishment of a Human Cell-Based In Vitro Test System for Investigating and Evaluating Cytotoxic and
Anja Wolff1, Jessica Reinert1, Silke Steinborn1
1Unit 4.II.2 Bioaerosols, Federal Institute of Occupational Safety and Health, Berlin, Germany.
Abstract:
Mold growth in damp indoor environments is a widespread problem associated with respiratory health effects, though causal mechanisms remain incompletely understood. We established a human cell-based in vitro test system using bronchial epithelial cells (NuLi-1) and macrophages (THP-1) to assess inhalation toxicity and immunomodulatory effects of indoor molds, focusing on cellular responses and mycotoxin patterns from contaminated building materials. The system integrates endpoints and impedance-based real-time cell viability analysis, quantification of released cytokines, and targeted mycotoxin profiling for comprehensive toxicological insights. Methanolic extracts from plasterboard, woodchip wallpaper, and malt extract agar (sterile PBS-treated or contaminated with Alternaria botrytis, Aspergillus versicolor, Penicillium chrysogenum, and Stachybotrys chartarum) were prepared via direct extraction of the material or surface swabs of the contaminated material. The system detected fungus- and material-specific effects, with pronounced cytotoxicity and immunomodulation, especially in Stachybotrys chartarum-contaminated samples. This material increased cytotoxicity in both tested cell types, decreased GM-CSF release from NuLi-1 cells, as well as increased interleukin-1β release from THP-1 macrophages. Therefore, mycotoxin composition and concentration were analyzed and confirmed elevated levels of the mycotoxins Roridin L2, Verrucarin J, and Stachybotrylactam. These findings reveal a substantial inhalation toxicity potential from Stachybotrys chartarum-contaminated indoor materials, highlighting health risks in damp environments and necessitating further in vivo and epidemiological studies.
Insights
Indoor mold exposure, particularly from Stachybotrys chartarum, poses significant inhalation toxicity risks. This study developed a cell-based system to assess mold toxicity and mycotoxin effects on respiratory health.
Area of Science:
- Environmental Health
- Toxicology
- Cell Biology
Background:
- Indoor mold contamination is linked to respiratory issues, but mechanisms are unclear.
- Understanding mold toxicity is crucial for public health in damp environments.
Purpose of the Study:
- To develop and utilize a human cell-based in vitro system for assessing inhalation toxicity and immunomodulatory effects of indoor molds.
- To analyze cellular responses and mycotoxin patterns from mold-contaminated building materials.
Main Methods:
- Established a co-culture system with bronchial epithelial cells (NuLi-1) and macrophages (THP-1).
- Integrated real-time cell viability analysis, cytokine quantification, and mycotoxin profiling.
- Tested methanolic extracts from various building materials contaminated with common indoor molds.
Main Results:
- Detected fungus- and material-specific toxicological effects.
- Stachybotrys chartarum-contaminated samples showed significant cytotoxicity and immunomodulation.
- Confirmed elevated levels of specific mycotoxins (Roridin L2, Verrucarin J, Stachybotrylactam) in contaminated materials.
Conclusions:
- Stachybotrys chartarum-contaminated indoor materials exhibit substantial inhalation toxicity.
- Findings highlight health risks associated with damp indoor environments.
- Further in vivo and epidemiological studies are warranted to confirm these health risks.
