Analyzing Murine Microglial Cell Response to Different Types of Particulate Matter

Abstract

Alzheimer’s and other neurodegenerative diseases affect nearly 24.3 million individuals globally— a number that will likely increase with the rising world population [1]. Studies conducted in Mexico City, Mexico suggest there may be a link between air pollution and chronic inflammation in the brain [4;5]. Research by Lillian Calderón‐Garcidueñas et al. showed that dogs and humans living in Mexico City, Mexico, a city with high amounts of air pollution, developed higher amounts of inflammation in the brain compared to humans and dogs residing in more rural areas [2;3]. Two hallmarks of Alzheimer’s and other related neurodegenerative diseases are the accumulation of β‐amyloid plaques and oxidative stress resulting in apoptosis. Exposing diesel exhaust particulate (DEP) from an industrial forklift engine (NIST, Boulder, CO) and urban dust (UD) filtered from Washington, D.C. (NIST, Boulder, CO) to cultured murine microglial cells (CRL‐2467; ATCC, Manassas, VA), we measured for the activation, production of reactive oxygen species (ROS), and apoptosis in microglial cells. We measured for caspase‐3 activity using a Caspase‐3/CPP32 Colorimetric Kit (Life Technologies, Fredrick, MD), activation by measuring for TNF‐α using mouse tumor necrosis factor α Elisa Kit (Sigma Aldrich, St. Louis, MO), and measured for ROS using an amplex red assay (ThermoFisher Scientific, Minneapolis, MN). We found a possible increase in TNF‐α production in cells cultured with DEP and no increase in ROS in our cells cultured with neither DEP nor UD.Support or Funding InformationSouth Dakota Biomedical Research Infrastructure Network