Abstract

Abstract

Amiodarone Effects on Phospholipidosis in NR8383 Lung Macrophages: A Multimodal Analysis Using SIMS and Fluorescence Microscopy

Matin Bastani

Abstract: Cationic amphiphilic Ddugs (CADs) constitute an intriguing group of FDA-approved pharmaceuticals that have the capability to accumulate within human cells, influencing their functions and structural integrity.1 Our research focuses on gaining a deeper understanding of the accumulation of amiodarone, an antiarrhythmic drug classified as a CAD, within a specific model system, NR8383 lung macrophages. To achieve this, we employed advanced techniques such as fluorescent microscopy and time-of-flight secondary ion mass spectrometry (ToF-SIMS) to shed light on this phenomenon. Our investigation is particularly concerned with phospholipidosis, a condition characterized by abnormal lipid build-up in cellular membranes induced by CADs.2 By characterizing the accumulation of amiodarone within individual cells, our research aims to unveil the underlying mechanisms. Two fundamental factors, drug dosage and incubation time, were central to our study. By exploring the effects of varying concentrations and exposure durations, we gained valuable insights into the dynamics of drug uptake and accumulation.
We utilized the state-of-the-art PHI nanoTOF II TOF-SIMS instrument to image individual cells, dosed and un-dosed, and compared their biochemical signatures.
Fluorescence microscopy was used to characterize the frequency and size of lysosomes, as well as the total cell size and shape in each cell. Despite the cells being monoclonal and cultured under consistent environmental conditions, we observed a significant variation in their response to amiodarone. By understanding the factors underlying this variation within a cell population, we can gain insights into the broader context of drug-cellular interactions and identify toxicological indicators.