High-content analysis for drug delivery and nanoparticle applications

Abstract

Publication information Drug Discovery Today, 20 (8): 942-957Publisher Elsevier Item record/more information http://hdl.handle.net/10197/6636 Publisher's statementThis is the author's version of a work that was accepted for publication in Drug Discovery Today. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Drug Discovery Today, 20 (8), (2015 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Page 1 of 26A c c e p t e d M a n u s c r i p t Sally-Ann CryanSally-Ann Cryan has a pharmacy degree and a PhD in pharmaceutics ( Jeremy SimpsonJeremy Simpson carried out his PhD at the University of Warwick, followed postdoctoral work at the Scripps Research Institute in San Diego, and the Imperial Cancer Research Fund in London. After 9 years as a staff member at the European Molecular Biology Laboratory (EMBL) in Heidelberg (Germany), he was appointed as professor of cell biology at UCD, in 2008. He currently applies high-throughput imaging technologies to study subcellular transport pathways and the internalization routes taken by nanoparticles in cells. He runs the UCD Cell Screening Laboratory and is the author of more than 80 publications.High-content analysis (HCA) provides quantitative multiparametric cellular fluorescence data. From its origins in discovery toxicology, it is now addressing fundamental questions in drug delivery. Nanoparticles (NPs), polymers, and intestinal permeation enhancers are being harnessed in drug delivery systems to modulate plasma membrane properties and the intracellular environment. Identifying comparative mechanistic cytotoxicity on sublethal events is crucial to expedite the development of such systems. NP uptake and intracellular routing pathways are also being dissected using chemical and genetic perturbations, with the potential to assess the intracellular fate of targeted and untargeted particles in vitro. As we discuss here, HCA is set to make a major impact in preclinical delivery research by elucidating the intracellular pathways of NPs and the in vitro mechanistic-based toxicology of formulation constituents. A brief recap of cytotoxicity assaysIn vitro cell-based assays have long been used to assess the cytotoxic effects of drug exposure [1]. Cells undergoing acute necrosis swell and lose metabolic capacity, thereby losing the capacity to maintain a barrier to the extracellular space and the ability ...

The Effects of Physicochemical Properties of Nanomaterials on Their Cellular Uptake In Vitro and In Vivo

“…Up to now, most studies of NMs cellular uptake are achieved based on in vitro cell models. Indeed, in vitro model possesses its advantages on straightforwardly observing the process of NMs uptake, accurately quantifying intracellular NMs, precisely controlling NMs uptake behaviors at the single cell level . However, the significant inconsistency between in vitro and in vivo microenvironments, in many cases, greatly discounts the value of in vitro models on guiding the research and application of NMs.…”

“…Along with the growing importance of the biomedical applications of NMs, in recent years, the studies aiming at NMs cellular uptake show an explosive growth . For example, developing new methods to quantify NMs cellular uptake has become an important research direction . High‐content analysis based fluorescence‐labeling technology provides quantitative multiparametric intracellular fluorescence data to analyze NMs uptake .…”

“…For example, developing new methods to quantify NMs cellular uptake has become an important research direction . High‐content analysis based fluorescence‐labeling technology provides quantitative multiparametric intracellular fluorescence data to analyze NMs uptake . Moreover, how to enhance NMs cellular uptake efficiency is another hot topic.…”

Emerging Trend of Advanced Image Analysis in Nanomedicine

“…with the help of fluorescence based on fully automated detection platforms capable of imaging thousands of cells at any time point and this has opened new avenues in understanding how NPs interact with cells or cause toxicity [18]. In high-content analysis (HCA) a mixture of various dyes is used to measure different parameters of cytotoxicity (e.g., mitochondrial membrane rupture, intracellular calcium levels, cell proliferation, etc.)…”

A High‐Throughput Automated Confocal Microscopy Platform for Quantitative Phenotyping of Nanoparticle Uptake and Transport in Spheroids

“…This contrasts with more considerable efforts made to characterize NP uptake and endocytic trafficking in cells growing in 2D . In order to enhance our understanding of NP‐spheroid interactions in both a quantitative and systematic way, potentially capturing NP events at the subcellular, cell and spheroid levels, high‐content screening (HCS) presents a uniquely suitable approach …”