In Vitro Multiparameter Assay Development Strategy toward Differentiating Macrophage Responses to Inhaled Medicines

Ewelina Hoffman, Abhinav Kumar, Varsha Kanabar, Matthew Arno, Lucas Preux, Val Millar, Clive Page, Helen Collins, Ian Mudway, Lea Ann Dailey, Ben Forbes

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)


Although foamy macrophages (FMΦ) are commonly observed during nonclinical development of medicines for inhalation, there are no accepted criteria to differentiate adaptive from adverse FMΦ responses in drug safety studies. The purpose of this study was to develop a multiparameter in vitro assay strategy to differentiate and characterize different mechanisms of drug-induced FMΦ. Amiodarone, staurosporine, and poly(vinyl acetate) nanoparticles were used to induce distinct FMΦ phenotypes in J774A.1 cells, which were then compared with negative controls. Treated macrophages were evaluated for morphometry, lipid accumulation, gene expression, apoptosis, cell activation, and phagocytosis. Analysis of vacuolization (number/area vacuoles per cell) and phospholipid content revealed inducer-dependent distinctive patterns, which were confirmed by electron microscopy. In contrast to the other inducers, amiodarone increased vacuole size rather than number and resulted in phospholipid accumulation. No pronounced dysregulation of transcriptional activity or apoptosis was observed in response to sublethal concentrations of all inducers. Functionally, FMΦ induction did not affect macrophage activation by lipopolysaccharide, but it reduced phagocytic capacity, with different patterns of induction, severity, and resolution observed with the different inducers. An in vitro multiparameter assay strategy is reported that successfully differentiates and characterizes mechanisms leading to FMΦ induction by different types of agents.
Original languageEnglish
Pages (from-to)2675-87
Number of pages13
JournalMolecular Pharmaceutics
Issue number8
Early online date19 May 2015
Publication statusPublished - 3 Aug 2015


  • Administration, Inhalation
  • Amiodarone
  • Animals
  • Biological Assay
  • Cell Differentiation
  • Cells, Cultured
  • Foam Cells
  • Lethal Dose 50
  • Macrophage Activation
  • Macrophages
  • Mice
  • Mice, Inbred BALB C
  • Nanoparticles
  • Polyvinyls
  • Staurosporine
  • Vacuoles
  • Journal Article
  • Research Support, Non-U.S. Gov't


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