Interlaboratory evaluation of a digital holographic microscopy-based assay for label-free in vitro cytotoxicity testing of polymeric nanocarriers.

Content Provider	Europe PMC
Author	Marzi, Anne Eder, Kai Moritz Barroso, Álvaro Wågbø, Ane Marit Mørch, Ýrr Hatletveit, Anne Rein Visnes, Torkild Schmid, Ruth B. Klinkenberg, Geir Kemper, Björn Schnekenburger, Jürgen
Abstract	State-of-the-art in vitro test systems for nanomaterial toxicity assessment are based on dyes and several staining steps which can be affected by nanomaterial interference. Digital holographic microscopy (DHM), an interferometry-based variant of quantitative phase imaging (QPI), facilitates reliable proliferation quantification of native cell populations and the extraction of morphological features in a fast and label- and interference-free manner by biophysical parameters. DHM therefore has been identified as versatile tool for cytotoxicity testing in biomedical nanotechnology. In a comparative study performed at two collaborating laboratories, we investigated the interlaboratory variability and performance of DHM in nanomaterial toxicity testing, utilizing complementary standard operating procedures (SOPs). Two identical custom-built off-axis DHM systems, developed for usage in biomedical laboratories, equipped with stage-top incubation chambers were applied at different locations in Europe. Temporal dry mass development, 12-h dry mass increments and morphology changes of A549 human lung epithelial cell populations upon incubation with two variants of poly(alkyl cyanoacrylate) (PACA) nanoparticles were observed in comparison to digitonin and cell culture medium controls. Digitonin as cytotoxicity control, as well as empty and cabazitaxel-loaded PACA nanocarriers, similarly impacted 12-h dry mass development and increments as well as morphology of A549 cells at both participating laboratories. The obtained DHM data reflected the cytotoxic potential of the tested nanomaterials and are in agreement with corresponding literature on biophysical and chemical assays. Our results confirm DHM as label-free cytotoxicity assay for polymeric nanocarriers as well as the repeatability and reproducibility of the technology. In summary, the evaluated DHM assay could be efficiently implemented at different locations and facilitates interlaboratory in vitro toxicity testing of nanoparticles with prospects for application in regulatory science.Graphical abstractSupplementary InformationThe online version contains supplementary material available at 10.1007/s13346-022-01207-5.
Related Links	https://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC9263039&blobtype=pdf
ISSN	2190393X
Journal	Drug Delivery and Translational Research [Drug Deliv Transl Res]
Volume Number	12
DOI	10.1007/s13346-022-01207-5
PubMed Central reference number	PMC9263039
Issue Number	9
PubMed reference number	35799027
e-ISSN	21903948
Language	English
Publisher	Springer US
Publisher Date	2022-07-08
Publisher Place	New York
Access Restriction	Open
Rights License	Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. © The Author(s) 2022
Subject Keyword	Digital holographic microscopy Quantitative phase imaging Label-free cytotoxicity testing Nanoparticles Interlaboratory comparison In vitro Regulatory science Technology transfer
Content Type	Text
Resource Type	Article
Subject	Pharmaceutical Science