A High-throughput Lung Air-blood Barrier Assay for Aerosol Toxicity Evaluation

SEONGBIN JO, Liang-Hsin Chen, Shuichi Takayama, Nga Lee Ng, Georgia Institute of Technology

     Abstract Number: 511
     Working Group: Health-Related Aerosols

Abstract
The development of new approach methodologies (NAMs) for inhalation toxicity testing is critical to address the ethical concerns and species-specific limitations associated with in vivo studies. However, current in vitro models often lack the physiological complexity or throughput necessary for comprehensive toxicity evaluation. To address this gap, we developed a high-throughput air-blood barrier array (ABBA) model incorporating lung epithelial (NCI-H441) and endothelial (HUVEC) cells under an air-liquid interface (ALI) condition to assess aerosol-induced lung injury and inflammation. To further enhance physiological relevance, human macrophage cells (differentiated THP-1 cells) were incorporated into the system to model the cytokine secretion as immune responses following destruction in barrier integrity. We demonstrated the capability of this novel platform through investigating the toxicity of secondary organic aerosols (SOA) derived from naphthalene and α-pinene, two representative precursors of anthropogenic and biogenic emissions. These SOA were generated under controlled atmospheric chamber conditions and have been intensively characterized for their cellular toxicity using murine macrophage models in 96-well formats in our previous studies. Building upon this foundation, the ABBA model was applied to evaluate how these SOA perturb the air-blood barrier integrity and induce inflammatory responses at the epithelial-endothelial interface in dose-response manners.