Ultrafine Black Carbon: A Cumulative Assessment on The Physiology of Lung - Derived Cells

Combustion-derived nanomaterials are noxious ultrafine (\u3c100 nm) aerosolized by-products generated by human activity. They pose threats to pulmonary health due to their small size, allowing them to penetrate deep into the alveolar regions causing detrimental responses downstream. Indeed, an examination between nanocarbon particle exposure and poor pulmonary health via cellular behavior was needed. I hypothesized that low-dose and long-term administrations of carbonaceous nanoparticles contribute to respiratory conditions by irritating lung-derived cells. Responses to ultrafine black carbon (UBC), a key component of airborne pollutants, by human lung A549 and murine lung LA4 epithelial cells, human peripheral-blood monocytes THP1, and murine macrophages RAW264.7 were investigated. These lines were chosen because epithelial cells and mononuclear cells (monocytes and macrophages) are the first to come in contact with inhaled particulate matter (PM) and contribute to stress and inflammation of pulmonary tissues. The cells were first plated on day zero and fed fresh UBC suspended in culture media on days one, four, and seven. The exposure regimen included three different low-level concentrations of UBC. On day ten, all cells were prepared for subsequent assays. Effects on cellular viability revealed that UBC was modestly cytotoxic while ATPase and esterase activity (termed enzyme vitality for the purpose of this study) were significantly diminished in a dose-dependent manner. Additionally, beta-galactosidase proportionally increased with UBC concentration compared to untreated cells, indicating that cellular senescence was promoted across all cell types. Multiple markers of oxidative stress were subsequently measured, showing non-uniform trends and displaying moderate accumulation of oxidative damage, contingent upon cell line and UBC treatment. However, a significant induction of inner mitochondrial membrane depolarization of human UBC-treated cells was observed; an aberrant cell-cycle progression was found across all cells. The implemented regimen of UBC exposure elicited an imbalance of free (reduced) glutathione, altered mitochondrial potential, and abnormal cell-cycle activity. This method demonstrated that modifications across both species induced changes to cellular fitness overall. The data supported that compounding nanosized black carbon exposure could negatively affect overall pulmonary cell health by distinctively modifying intracellular activity. Additionally, we further surmised that sustained exposure to nanocarbon could impair innate immunity. Therefore, reactive oxygen species (ROS), reactive nitrogen species (RNS), and cytokine levels upon continuous administrations to 3.0 μg/mL UBC after brief challenges to microbial proxies were assessed; on day 9, the cells were succumbed to three different microbial insults (LPS, Poly I:C, and Zymosan). On day 10, measurements demonstrated a moderate accumulation of oxidative stress, conditional to each cell type. Thus, UBC plus microbe approach triggered pro-oxidant levels and cytokine production across different cell types and diminished phagocytic response by murine macrophages. Based on the results, ultrafine-PM exposure may impair inflammatory function in response to pathogenic infections. In summation, our findings indicated that additive nanosized UBC in combination with microbe-like challenges may compromise cellular behavior, suggesting potential ways by which inhalation of ultrafine-PM may contribute to or exacerbate poor pulmonary health