Microfluidics as a Novel Tool for Biological and Toxicological Assays in Drug Discovery Processes: Focus on Microchip Electrophoresis

Abstract

The last decades of biological, toxicological, and pharmacological research have deeply changed the way researchers select the most appropriate ‘pre-clinical model’. The absence of relevant animal models for many human diseases, as well as the inaccurate prognosis coming from ‘conventional’ pre-clinical models, are among the major reasons of the failures observed in clinical trials. This evidence has pushed several research groups to move more often from a classic cellular or animal modeling approach to an alternative and broader vision that includes the involvement of microfluidic-based technologies. The use of microfluidic devices offers several benefits including fast analysis times, high sensitivity and reproducibility, the ability to quantitate multiple chemical species, and the simulation of cellular response mimicking the closest human in vivo milieu. Therefore, they represent a useful way to study drug–organ interactions and related safety and toxicity, and to model organ development and various pathologies ‘in a dish’. The present review will address the applicability of microfluidic-based technologies in different systems (2D and 3D). We will focus our attention on applications of microchip electrophoresis (ME) to biological and toxicological studies as well as in drug discovery and development processes. These include high-throughput single-cell gene expression profiling, simultaneous determination of antioxidants and reactive oxygen and nitrogen species, DNA analysis, and sensitive determination of neurotransmitters in biological fluids. We will discuss new data obtained by ME coupled to laser-induced fluorescence (ME-LIF) and electrochemical detection (ME-EC) regarding the production and degradation of nitric oxide, a fundamental signaling molecule regulating virtually every critical cellular function. Finally, the integration of microfluidics with recent innovative technologies—such as organoids, organ-on-chip, and 3D printing—for the design of new in vitro experimental devices will be presented with a specific attention to drug development applications. This ‘composite’ review highlights the potential impact of 2D and 3D microfluidic systems as a fast, inexpensive, and highly sensitive tool for high-throughput drug screening and preclinical toxicological studies.

RETRACTED: The Enhanced Cytotoxic and Pro-Apoptotic Effects of Optimized Simvastatin-Loaded Emulsomes on MCF-7 Breast Cancer Cells

“…Furthermore, in order to improve the above-mentioned activities of our formulation, alternative and innovative methods to determine the kinetic release of the content from the EMLs, such as USP 4 [62], will be investigated. Our results also make the base for the investigation of our optimized formula in 3D/organoid systems [63] as well as in vivo.…”

Microfluidic Tools for Enhanced Characterization of Therapeutic Stem Cells and Prediction of Their Potential Antimicrobial Secretome

“…Both organoids and spheroids can be formed by primary cells and stem cells. The combination of these 3D cell cultures with microfluidic systems, such as a sophisticated organ-on-a-chip [180] will develop more and more relevant in vitro pre-clinical models [181] to study human pathology, accelerate discovery and screening of new drugs, or biologics such as new host defense peptides [45], and evaluate treatments. The limitation of flow cytometric analysis is the inevitable disruption of spatial spheroid organization by cell dissociation and loss of their whole properties as unique multicellular elements.…”

The Role of Dielectrophoresis for Cancer Diagnosis and Prognosis

“…One of the main aims of biological and biochemical studies is to understand the mechanisms characterizing a single cell, which is the minimal functional unit of life. In this context, the application of microfluidic-based devices offers numerous advantages, including the rapid and simultaneous separation and quantification of multiple chemical species, high sensitivity and reproducibility, and the possibility to study the heterogeneity of cell populations [ 62 ], the last being an extremely important factor to be considered for the analysis of CTCs. The use of these devices is also accompanied by a very short analysis time and gives the opportunity to use different detection platforms, such as fluorescence and electrochemical detection, just to name two, making it useful for high-throughput single-cell analysis.…”