Microglia as a Surrogate Biosensor to Determine Nanoparticle Neurotoxicity.
Nanoparticles found in air pollutants can alter neurotransmitter profiles, increase neuroinflammation, and alter brain function. Therefore, the assay described here will aid in elucidating the role of microglia in neuroinflammation and neurodegenerative diseases. The use of microglia, resident immune cells of the brain, as a surrogate biosensor provides novel insight into how inflammatory responses mediate neuronal insults. Here, we utilize an immortalized murine microglial cell line, designated BV2, and describe a method for nanoparticle exposure using silver nanoparticles (AgNPs) as a standard. We describe how to expose microglia to nanoparticles, how to remove nanoparticles from supernatant, and how to use supernatant from activated microglia to determine toxicity, using hypothalamic cell survival as a measure. Following AgNP exposure, BV2 microglial activation was validated using a tumor necrosis factor alpha (TNF-α) enzyme linked immunosorbent assay (ELISA). The supernatant was filtered to remove the AgNP and to allow cytokines and other secreted factors to remain in the conditioned media. Hypothalamic cells were then exposed to supernatant from AgNP activated microglia and survival of neurons was determined using a resazurin-based fluorescent assay. This technique is useful for utilizing microglia as a surrogate biomarker of neuroinflammation and determining the effect of neuroinflammation on other cell types.
Minneapolis Veterans Affairs Health Care System; Department of Food Science and Nutrition, University of Minnesota.
Cayla M
CM
Duffy
Minneapolis Veterans Affairs Health Care System.
Shihab
S
Ahmed
Minneapolis Veterans Affairs Health Care System; Department of Food Science and Nutrition, University of Minnesota; Biomedical Informatics and Computational Biology Program, University of Minnesota.
Ce
C
Yuan
Minneapolis Veterans Affairs Health Care System.
Vijayakumar
V
Mavanji
Minneapolis Veterans Affairs Health Care System; Department of Food Science and Nutrition, University of Minnesota.
Joshua P
JP
Nixon
Minneapolis Veterans Affairs Health Care System; Department of Food Science and Nutrition, University of Minnesota; Minnesota Obesity Center, University of Minnesota; butte017@umn.edu.
Tammy
T
Butterick
D000818:Animals; D015374:Biosensing Techniques; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D007031:Hypothalamus; D051379:Mice; D017628:Microglia; D053758:Nanoparticles / Q000633:toxicity*; D018675:Toxicity Tests; D014409:Tumor Necrosis Factor-alpha
D014409:Tumor Necrosis Factor-alpha