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Cytotoxicity analysis of biomass combustion particles in human pulmonary alveolar epithelial cells on an air–liquid interface/dynamic culture platform
Particle and Fibre Toxicology ( IF 10 ) Pub Date : 2021-08-21 , DOI: 10.1186/s12989-021-00426-x
Shaorui Ke 1, 2 , Qi Liu 2 , Xinlian Zhang 2 , Yuhan Yao 2 , Xudong Yang 3 , Guodong Sui 2, 4
Affiliation  

Exposure to indoor air pollution from solid fuel combustion is associated with lung diseases and cancer. This study investigated the cytotoxicity and molecular mechanisms of biomass combustion-derived particles in human pulmonary alveolar epithelial cells (HPAEpiC) using a platform that combines air–liquid interface (ALI) and dynamic culture (DC) systems. HPAEpiC were cultured on the surface of polycarbonate (PC) membranes on the ALI–DC platform. The cells were sprayed with an aerosolized solution of biomass combustion soluble constituents (BCSCs) and simultaneously nourished with culture medium flowing beneath the permeable PC membranes. The ALI–DC method was compared with the traditional submerged culture approach. BCSC particle morphology and dosages deposited on the chip were determined for particle characterization. Flow cytometry, scanning electron microscopy, and transmission electron microscopy were used to investigate the apoptosis rate of HPAEpiC and changes in the cell ultrastructure induced by BCSCs. Additionally, the underlying apoptotic pathway was examined by determining the protein expression levels by western blotting. Scanning electron microscope images demonstrated that the sample processing and delivering approach of the ALI–DC platform were suitable for pollutant exposure. Compared with the submerged culture method, a significant decline in cell viability and increase in apoptosis rate was observed after BCSC exposure on the ALI–DC platform, indicating that the ALI–DC platform is a more sensitive system for investigating cytotoxicity of indoor air pollutants in lung cells. The morphology and ultrastructure of the cells were damaged after exposure to BCSCs, and the p53 pathway was activated. The Bcl-2/Bax ratio was reduced, upregulating caspase-9 and caspase-3 expression and subsequently inducing apoptosis of HPAEpiC. The addition of N-acetyl cysteine antioxidant significantly alleviated the cytotoxicity induced by BCSCs. A novel ALI–DC platform was developed to study the cytotoxicity of air pollutants on lung cells. Using the platform, we demonstrated that BCSCs could damage the mitochondria, produce reactive oxygen species, and activate p53 in HPAEpiC, ultimately inducing apoptosis.

中文翻译:

气液界面/动态培养平台上人肺泡上皮细胞中生物质燃烧颗粒的细胞毒性分析

暴露于固体燃料燃烧造成的室内空气污染与肺部疾病和癌症有关。本研究使用结合气液界面 (ALI) 和动态培养 (DC) 系统的平台,研究了人肺泡上皮细胞 (HPAEpiC) 中生物质燃烧衍生颗粒的细胞毒性和分子机制。HPAEpiC 在 ALI-DC 平台上的聚碳酸酯 (PC) 膜表面上培养。用生物质燃烧可溶性成分 (BCSC) 的雾化溶液喷洒细胞,同时用在可渗透 PC 膜下方流动的培养基滋养。ALI-DC 方法与传统的淹没培养方法进行了比较。BCSC 颗粒形态和沉积在芯片上的剂量被确定用于颗粒表征。流式细胞术,扫描电子显微镜和透射电子显微镜用于研究HPAEpiC的凋亡率和BCSCs诱导的细胞超微结构的变化。此外,通过蛋白质印迹确定蛋白质表达水平来检查潜在的细胞凋亡途径。扫描电子显微镜图像表明,ALI-DC 平台的样品处理和输送方法适用于污染物暴露。与浸没培养方法相比,在 ALI-DC 平台上暴露 BCSC 后观察到细胞活力显着下降和细胞凋亡率增加,表明 ALI-DC 平台是研究室内空气污染物细胞毒性的更敏感系统。肺细胞。暴露于 BCSCs 后细胞的形态和超微结构受损,p53 通路被激活。Bcl-2/Bax 比率降低,上调 caspase-9 和 caspase-3 表达,随后诱导 HPAEpiC 细胞凋亡。N-乙酰半胱氨酸抗氧化剂的加入显着减轻了BCSCs诱导的细胞毒性。开发了一种新的 ALI-DC 平台来研究空气污染物对肺细胞的细胞毒性。使用该平台,我们证明了 BCSCs 可以损伤线粒体,产生活性氧,并激活 HPAEpiC 中的 p53,最终诱导细胞凋亡。开发了一种新的 ALI-DC 平台来研究空气污染物对肺细胞的细胞毒性。使用该平台,我们证明了 BCSCs 可以损伤线粒体,产生活性氧,并激活 HPAEpiC 中的 p53,最终诱导细胞凋亡。开发了一种新的 ALI-DC 平台来研究空气污染物对肺细胞的细胞毒性。使用该平台,我们证明了 BCSCs 可以损伤线粒体,产生活性氧,并激活 HPAEpiC 中的 p53,最终诱导细胞凋亡。
更新日期:2021-08-21
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