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Quantitative Phase Imaging as Sensitive Screening Method for Nanoparticle-Induced Cytotoxicity Assessment.

Anne Marzi1, Kai Moritz Eder1, Álvaro Barroso1

  • 1Biomedical Technology Center, University of Muenster, Mendelstraße 17, D-48149 Muenster, Germany.

Cells
|April 26, 2024
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Summary

Digital holographic microscopy (DHM) offers a more sensitive, label-free method for assessing nanoparticle cytotoxicity. This technique surpasses traditional biochemical assays in detecting cellular effects, aiding in standardized nanoparticle testing.

Keywords:
digital holographic microscopyin vitrolabel-free cytotoxicity testingnanoparticlesquantitative phase imaging

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Area of Science:

  • Nanotechnology
  • Toxicology
  • Biophysics

Background:

  • Assessing nanoparticle cytotoxicity is complex due to a lack of standardized testing guidelines.
  • Nanoparticles' unique properties can interfere with conventional biochemical assays, necessitating multiple testing methods.
  • There is a need for particle-unaffected methods that integrate molecular techniques for reliable nanoparticle assessment.

Purpose of the Study:

  • To investigate the capability of Digital Holographic Microscopy (DHM) in a multimodal approach for cytotoxicity assessment.
  • To compare DHM-detected cellular effects with downstream biochemical assays for nanoparticle toxicity evaluation.
  • To determine the sensitivity of DHM versus metabolic activity and lactate dehydrogenase assays in detecting nanoparticle-induced cytotoxicity.

Main Methods:

  • Utilized quantitative DHM phase contrast to measure dry mass changes in RAW 264.7 macrophages and NIH-3T3 fibroblasts after exposure to poly(alkyl cyanoacrylate) nanoparticles.
  • Compared DHM results with cytotoxic (digitonin) and medium controls.
  • Assessed cell viability using a WST-8 metabolic activity assay and cell death via lactate dehydrogenase (LDH) release assay in supernatants.

Main Results:

  • DHM demonstrated higher sensitivity in detecting the effects of nanoparticles on the tested cell lines compared to the WST-8 and LDH assays.
  • Quantitative DHM effectively measured dry mass increase as an indicator of cellular response to nanoparticles.
  • The average half-maximal effective concentration (EC50) determined by DHM was more sensitive to nanocarrier effects.

Conclusions:

  • Digital holographic microscopy is a promising label-free technique for sensitive and comprehensive nanoparticle cytotoxicity assessment.
  • DHM's minimal sample interaction allows for downstream analyses, supporting a multimodal testing strategy.
  • Integrating DHM into nanoparticle testing routines can enhance reliability and standardization, overcoming limitations of current biochemical assays.