Development of bioassays for gaseous chemicals using air-liquid interface culture of human lung epithelial cells

利用人肺上皮细胞气液界面培养物开发气态化学物质生物测定法

基本信息

批准号：
13480163
负责人：
SAKAI Yasuyuki
金额：
$ 7.55万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13480163/
关键词：
Alveolus Air way ir-liquid interface culture damage to the lung intake Suspended particular matter Cytochrome P450 Numerical simulation 浮遊粒子状物体肺胞上皮肺気道上皮バイオアッセイ有機溶媒

项目摘要

To establish a series of toxicitiy evaluation systems for chemicals or environmental samples in gas phase, we employed air-liquid interface culture (ALIC) of human bronchial (air way) and alveolus epithelial cells as a basic culture method, and investigated their applications to volatile organic compounds and suspended paniculate matter (SPM).First, using a human bronchial epithelial cell line, Calu-3, we developed a simple closed system for direct gas exposure, and tested the toxicity expression of Calu-3 to several organic compounds during 48 hours of loading. The toxicity in ALIC exposure was higher than that in conventional exposure in the liquid phase. The reason was largely explained by numerical estimation that chemical concentration on the cell surface in the liquid culture is lower than that in ALIC culture, in the cause of the diffusion process of molecules in the surface liquid layer. These results indicate that basic concept of the combination of ALIC of lung cells and a si … More mple closed system is promising as a testing cytotoxicrly to gaseous compounds.Second, in vivo mimicking exposure of actual SPM samples to human lung alveolar epithelia was achieved using an air-liquid interface culture of human lung alveolar A549 cells, resulting in successful observation of the toxicity in terms of the cell survivability after 48 hours exposure. In addition, an integrated use of the ALIC-based A549 cell assay for general toxicity and the EROD-based Hep G2 cell assay for PAHs was recommended to evaluate the primary toxicity of SPM. We thereby understood the permeation kinetics of SPM toxicity across the alveolar epithelia in terms of the EROD capacity of Hep G2 cells, and were able to obtain basic information on SPM toxicity in humans.Third, we developed a numerical simulation model that describes elution of chemicals initially adsorbed onto SPM samples, intake and accumulation of these chemicals into alveolus cell layer, and their final permeation to the systemic blood circulation. By focusing on the EROD activity of SPM samples, we quantified the comprehensive toxicity of the relevant samples and used these toxicitis in determining parameters involved in these numerical simulations. In addition, we extended the numerical models in actual situations that reflect low-dose and long-term exposure of SPM to humans across the very thin layer of the alveolar tissue. Less

建立一系列人支气管（空气方法）中气相的化学物质或环境样品的毒性评估系统，作为基本培养方法，并研究了它们在挥发性有机化合物和悬浮化合物中的应用，。在差异液体层的原因使用人lungarar A549的前面培养物来实现人类肺肺泡上皮的实际SPLE，从而成功地观察到了48小时后的细胞生存能力的毒性。建议使用基于EROD的HEP G2细胞测定PAHS来评估SPM的初级。。在实际情况下，SPM在肺泡组织的层中向人类长期暴露。