Advances in Microtechnology for Improved Cytotoxicity Assessment

In vitro cytotoxicity testing is essential in the pharmaceutical and environmental industry to study the effects of potential harmful compounds for human health. Classical assays present several disadvantages: they are commonly based on live-death labelling, are highly time consuming and/or require skilled personnel to be performed. The current trend is to reduce the number of required cells and the time during the analysis, while increasing the screening capability and the accuracy and sensitivity of the assays, aiming single cell resolution. Microfabrication and surface engineering are enabling novel approaches for cytotoxicity assessment, offering high sensitivity and the possibility of automation in order to minimize user intervention. This review aims to overview the different microtechnology approaches available in this field, focusing on the novel developments for high-throughput, dynamic and real time screening of cytotoxic compounds.

[1]  U. Liebel,et al.  Superhydrophobic-superhydrophilic micropatterning: towards genome-on-a-chip cell microarrays. , 2011, Angewandte Chemie.

[2]  Harry J. Levinson,et al.  Principles of Lithography , 2001 .

[3]  S. Ingebrandt,et al.  Electrical cell-substrate impedance sensing with field-effect transistors is able to unravel cellular adhesion and detachment processes on a single cell level. , 2015, Lab on a chip.

[4]  R. Ismagilov,et al.  Detecting bacteria and determining their susceptibility to antibiotics by stochastic confinement in nanoliter droplets using plug-based microfluidics. , 2008, Lab on a chip.

[5]  Martha L. Bulyk,et al.  Quantifying DNA–protein interactions by double-stranded DNA arrays , 1999, Nature Biotechnology.

[6]  M Stubenrauch,et al.  Cell cultures in microsystems: Biocompatibility aspects , 2011, Biotechnology and bioengineering.

[7]  R Puers,et al.  Digital microfluidics for time-resolved cytotoxicity studies on single non-adherent yeast cells. , 2015, Lab on a chip.

[8]  M. Bélanger,et al.  Hemocompatibility, biocompatibility, inflammatory and in vivo studies of primary reference materials low-density polyethylene and polydimethylsiloxane: a review. , 2001, Journal of biomedical materials research.

[9]  M. Stoddart.  Cell viability assays: introduction. , 2011, Methods in molecular biology.

[10]  Bifeng Liu,et al.  Microfluidic chip electrophoresis for biochemical analysis. , 2019, Journal of separation science.

[11]  Fangjuan Zhang,et al.  Three-gradient constructions in a flow-rate insensitive microfluidic system for drug screening towards personalized treatment. , 2020, Talanta.

[12]  L. Basabe‐Desmonts,et al.  Individual platelet adhesion assay: measuring platelet function and antiplatelet therapies in whole blood via digital quantification of cell adhesion. , 2013, Analytical chemistry.

[13]  M. Lutolf,et al.  Hydrogel microfluidics for the patterning of pluripotent stem cells , 2014, Scientific Reports.

[14]  Bingcheng Lin,et al.  Characterization of drug metabolites and cytotoxicity assay simultaneously using an integrated microfluidic device. , 2009, Lab on a chip.

[15]  C. Antczak,et al.  Cell viability assessment: toward content-rich platforms , 2010, Expert opinion on drug discovery.

[16]  Seunghun Hong,et al.  Reusable floating-electrode sensor for the quantitative electrophysiological monitoring of a nonadherent cell. , 2014, ACS nano.

[17]  Birgit Kersten,et al.  Multiplex approaches in protein microarray technology , 2005, Expert review of proteomics.

[18]  D. Beebe,et al.  Physics and applications of microfluidics in biology. , 2002, Annual review of biomedical engineering.

[19]  G. Lowry,et al.  Towards a definition of inorganic nanoparticles from an environmental, health and safety perspective. , 2009, Nature nanotechnology.

[20]  Probal Banerjee,et al.  Core-shell hybrid nanogels for integration of optical temperature-sensing, targeted tumor cell imaging, and combined chemo-photothermal treatment. , 2010, Biomaterials.

[21]  Mickael Guedj,et al.  Analysis of drug combinations: current methodological landscape , 2015, Pharmacology research & perspectives.

[22]  A. Zeng,et al.  Microtechnology meets systems biology: the small molecules of metabolome as next big targets. , 2010, Journal of biotechnology.

[23]  Y. Liu,et al.  An automated and portable microfluidic chemiluminescence immunoassay for quantitative detection of biomarkers. , 2017, Lab on a chip.

[24]  Su Hyun Jung,et al.  Robust chemical bonding of PMMA microfluidic devices to porous PETE membranes for reliable cytotoxicity testing of drugs. , 2019, Lab on a chip.

[25]  Pascal Colpo,et al.  Assessment of cytotoxicity by impedance spectroscopy. , 2007, Biosensors & bioelectronics.

[26]  Rashid Bashir,et al.  Reliable fabrication method of transferable micron scale metal pattern for poly(dimethylsiloxane) metallization. , 2006, Lab on a chip.

[27]  P. Levkin,et al.  Evaluation of the Droplet-Microarray Platform for High-Throughput Screening of Suspension Cells , 2017, SLAS technology.

[28]  Patrycja Ciosek,et al.  Flow-through sensor array applied to cytotoxicity assessment in cell cultures for drug-testing purposes. , 2014, Biosensors & bioelectronics.

[29]  Martin C. Hartel,et al.  Microphysiological Systems: Next Generation Systems for Assessing Toxicity and Therapeutic Effects of Nanomaterials. , 2019, Small methods.

[30]  C. Lim,et al.  Microfluidics for cell sorting and single cell analysis from whole blood. , 2018, Methods in cell biology.

[31]  Qiang Gao,et al.  Label-free electrochemical DNA biosensor array for simultaneous detection of the HIV-1 and HIV-2 oligonucleotides incorporating different hairpin-DNA probes and redox indicator. , 2010, Biosensors & bioelectronics.

[32]  N. Voelcker,et al.  The Use of Microfluidics in Cytotoxicity and Nanotoxicity Experiments , 2017, Micromachines.

[33]  Dayu Liu,et al.  Fabrication of a microfluidic chip containing dam, weirs and gradient generator for studying cellular response to chemical modulation , 2009 .

[34]  Fuqiang Nie,et al.  Non-dye cell viability monitoring by using pH-responsive inverse opal hydrogels. , 2018, Journal of materials chemistry. B.

[35]  K. Savolainen,et al.  Nanomaterial exposure, toxicity, and impact on human health. , 2018, Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology.

[36]  G. Luo,et al.  Single-Cell-Arrayed Agarose Chip for in Situ Analysis of Cytotoxicity and Genotoxicity of DNA Cross-Linking Agents. , 2016, Analytical chemistry.

[37]  H. Lashuel,et al.  Amyloid single-cell cytotoxicity assays by nanomotion detection , 2017, Cell Death Discovery.

[38]  Andrew J. Capel,et al.  3D printing for chemical, pharmaceutical and biological applications , 2018, Nature Reviews Chemistry.

[39]  J. Irelan,et al.  Rapid and Quantitative Assessment of Cell Quality, Identity, and Functionality for Cell-Based Assays Using Real-Time Cellular Analysis , 2011, Journal of biomolecular screening.

[40]  Oliver Lieleg,et al.  A microfluidics approach to study the accumulation of molecules at basal lamina interfaces. , 2015, Lab on a chip.

[41]  L. Basabe‐Desmonts,et al.  Extracellular matrix protein microarray-based biosensor with single cell resolution: Integrin profiling and characterization of cell-biomaterial interactions , 2019, Sensors and Actuators B: Chemical.

[42]  D. Grasso,et al.  Flow cytometry. , 1998, Methods in molecular medicine.

[43]  Gang-Yu Liu,et al.  Cell tracing dyes significantly change single cell mechanics. , 2009, The journal of physical chemistry. B.

[44]  A. Nelson,et al.  Substituents modulate biphenyl penetration into lipid membranes. , 2017, Biochimica et biophysica acta. Biomembranes.

[45]  L. Liz‐Marzán,et al.  Tunable Nanoparticle and Cell Assembly Using Combined Self‐Powered Microfluidics and Microcontact Printing , 2016 .

[46]  A K Uguz,et al.  Screening applications in drug discovery based on microfluidic technology. , 2016, Biomicrofluidics.

[47]  W. Bentley,et al.  Selective assembly and functionalization of miniaturized redox capacitor inside microdevices for microbial toxin and mammalian cell cytotoxicity analyses. , 2018, Lab on a Chip.

[48]  Fernando Benito-Lopez,et al.  Microtechnologies for Cell Microenvironment Control and Monitoring , 2017, Micromachines.

[49]  Hitoshi Shiku,et al.  Facile and rapid generation of 3D chemical gradients within hydrogels for high-throughput drug screening applications. , 2014, Biosensors & bioelectronics.

[50]  Seunghun Hong,et al.  Quantitative electrophysiological monitoring of anti-histamine drug effects on live cells via reusable sensor platforms. , 2017, Biosensors & bioelectronics.

[51]  S. Feinstein,et al.  Kinetic cell-based morphological screening: prediction of mechanism of compound action and off-target effects. , 2009, Chemistry & biology.

[52]  Ying Zhu,et al.  Cell-based drug combination screening with a microfluidic droplet array system. , 2013, Analytical chemistry.

[53]  Ian M. Kennedy,et al.  New Approach to Investigate the Cytotoxicity of Nanomaterials Using Single Cell Mechanics , 2014, The journal of physical chemistry. B.

[54]  Yanhui Hu,et al.  Next generation high density self assembling functional protein arrays , 2008, Nature Methods.

[55]  U. Demirci,et al.  Controlled viable release of selectively captured label-free cells in microchannels. , 2011, Lab on a chip.

[56]  Michael J Taussig,et al.  Printing protein arrays from DNA arrays , 2008, Nature Methods.

[57]  Xiao Xu,et al.  The xCELLigence system for real-time and label-free monitoring of cell viability. , 2011, Methods in molecular biology.

[58]  Ruili Huang,et al.  A bioluminescent cytotoxicity assay for assessment of membrane integrity using a proteolytic biomarker. , 2008, Toxicology in vitro : an international journal published in association with BIBRA.

[59]  D. Weitz,et al.  Droplet microfluidics: A tool for biology, chemistry and nanotechnology , 2016 .

[60]  D. Beebe,et al.  The present and future role of microfluidics in biomedical research , 2014, Nature.

[61]  Jie Wang,et al.  Microfluidic synthesis of barcode particles for multiplex assays. , 2015, Small.

[62]  Aysun Adan,et al.  Flow cytometry: basic principles and applications , 2017, Critical reviews in biotechnology.

[63]  Fernanda H Sakamoto,et al.  Introduction to confocal microscopy. , 2012, The Journal of investigative dermatology.

[64]  G. Barratt,et al.  A cell impedance‐based real‐time in vitro assay to assess the toxicity of amphotericin B formulations , 2017, Toxicology and applied pharmacology.

[65]  A. Nelson,et al.  Freezing or wrapping: the role of particle size in the mechanism of nanoparticle-biomembrane interaction. , 2012, Langmuir : the ACS journal of surfaces and colloids.

[66]  R. Horváth,et al.  Label-free optical biosensor for real-time monitoring the cytotoxicity of xenobiotics: A proof of principle study on glyphosate. , 2018, Journal of hazardous materials.

[67]  Qionglin Liang,et al.  Simultaneous Assay of Oxygen-Dependent Cytotoxicity and Genotoxicity of Anticancer Drugs on an Integrated Microchip. , 2018, Analytical chemistry.

[68]  L. Basabe‐Desmonts,et al.  Optical Single Cell Resolution Cytotoxicity Biosensor Based on Single Cell Adhesion Dot Arrays. , 2020, Analytical chemistry.

[69]  J. Owen,et al.  High-throughput electrochemical sensing platform for screening nanomaterial-biomembrane interactions. , 2020, The Review of scientific instruments.

[70]  Kit S Lam,et al.  Microfluidic-Enabled Print-to-Screen Platform for High-Throughput Screening of Combinatorial Chemotherapy. , 2015, Analytical chemistry.

[71]  Stephan Gabos,et al.  Dynamic monitoring of cytotoxicity on microelectronic sensors. , 2005, Chemical research in toxicology.

[72]  Liang Guo,et al.  A PDMS-Based Integrated Stretchable Microelectrode Array (isMEA) for Neural and Muscular Surface Interfacing , 2013, IEEE Transactions on Biomedical Circuits and Systems.

[73]  S. J. Milne,et al.  Serum albumin enhances the membrane activity of ZnO nanoparticles. , 2013, Chemical communications.

[74]  Y. S. Zhang,et al.  Automated microfluidic platform of bead-based electrochemical immunosensor integrated with bioreactor for continual monitoring of cell secreted biomarkers , 2016, Scientific Reports.

[75]  Penny Nymark,et al.  In vitro and in vivo genotoxic effects of straight versus tangled multi-walled carbon nanotubes , 2016, Nanotoxicology.

[76]  M. Rigoulet,et al.  Microwell array integrating nanoelectrodes for coupled opto-electrochemical monitorings of single mitochondria. , 2019, Biosensors & bioelectronics.

[77]  Bengt Fadeel,et al.  The Right Stuff: On the Future of Nanotoxicology , 2019, Front. Toxicol..

[78]  T. Konry,et al.  Phenotypic drug profiling in droplet microfluidics for better targeting of drug-resistant tumors. , 2015, Lab on a chip.

[79]  Songyao Jiang,et al.  Machine learning-aided quantification of antibody-based cancer immunotherapy by natural killer cells in microfluidic droplets. , 2020, Lab on a chip.

[80]  Gabriel P López,et al.  Microfluidic cell sorting: a review of the advances in the separation of cells from debulking to rare cell isolation. , 2015, Lab on a chip.

[81]  P. Pinton,et al.  Selective modulation of subtype III IP3R by Akt regulates ER Ca2+ release and apoptosis , 2012, Cell Death and Disease.

[82]  M. Niwano,et al.  Stable lipid bilayers based on micro- and nano-fabrication , 2010 .

[83]  T. Hug,et al.  Biophysical methods for monitoring cell-substrate interactions in drug discovery. , 2003, Assay and drug development technologies.

[84]  Luke P. Lee,et al.  Nanoliter scale microbioreactor array for quantitative cell biology , 2006, Biotechnology and bioengineering.

[85]  D. Weitz,et al.  Single-cell analysis and sorting using droplet-based microfluidics , 2013, Nature Protocols.

[86]  Masahito Hosokawa,et al.  Microfluidic device with chemical gradient for single-cell cytotoxicity assays. , 2011, Analytical chemistry.

[87]  B. Chung,et al.  A microfluidic gradient device for drug screening with human iPSC-derived motoneurons. , 2020, The Analyst.

[88]  P. Levkin,et al.  Droplet‐Array (DA) Sandwich Chip: A Versatile Platform for High‐Throughput Cell Screening Based on Superhydrophobic–Superhydrophilic Micropatterning , 2015, Advanced materials.

[89]  Z. Brzózka,et al.  Synergistic effect of the combination therapy on ovarian cancer cells under microfluidic conditions. , 2020, Analytica chimica acta.

[90]  Gang-Yu Liu,et al.  Cell mechanics using atomic force microscopy-based single-cell compression. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[91]  Tobias Bollenbach,et al.  Antimicrobial interactions: mechanisms and implications for drug discovery and resistance evolution. , 2015, Current opinion in microbiology.

[92]  Bai Yang,et al.  Elliptical polymer brush ring array mediated protein patterning and cell adhesion on patterned protein surfaces. , 2013, ACS applied materials & interfaces.

[93]  Liwei Lin,et al.  3D printed three-flow microfluidic concentration gradient generator for clinical E. Coli-antibiotic drug screening , 2017, 2017 IEEE 30th International Conference on Micro Electro Mechanical Systems (MEMS).

[94]  Chee Meng Benjamin Ho,et al.  3D printed microfluidics for biological applications. , 2015, Lab on a chip.

[95]  Zhou Zhou,et al.  3D printed microfluidic chip for multiple anticancer drug combinations , 2018, Sensors and Actuators B: Chemical.

[96]  J. Mee.  The synergistic effect , 1965 .

[97]  C. Recordati,et al.  Tissue distribution and acute toxicity of silver after single intravenous administration in mice: nano-specific and size-dependent effects , 2015, Particle and Fibre Toxicology.

[98]  Mitsutoshi Nakajima,et al.  Recent lab‐on‐chip developments for novel drug discovery , 2017, Wiley interdisciplinary reviews. Systems biology and medicine.

[99]  Julian R. Jones,et al.  3D Printing of Biocompatible Supramolecular Polymers and their Composites. , 2016, ACS applied materials & interfaces.

[100]  Hongwei Ma,et al.  Surface engineering strategies for control of protein and cell interactions , 2004 .

[101]  Shinji Sugiura,et al.  Pressure‐driven perfusion culture microchamber array for a parallel drug cytotoxicity assay , 2008, Biotechnology and bioengineering.

[102]  Hongkai Wu,et al.  A prototypic microfluidic platform generating stepwise concentration gradients for real-time study of cell apoptosis. , 2010, Biomicrofluidics.

[103]  Sungbo Cho,et al.  Hydrogel-based diffusion chip with Electric Cell-substrate Impedance Sensing (ECIS) integration for cell viability assay and drug toxicity screening. , 2013, Biosensors & bioelectronics.

[104]  G. Whitesides,et al.  Soft Lithography. , 1998, Angewandte Chemie.

[105]  J. A. Maurer,et al.  Microcontact printing for creation of patterned lipid bilayers on tetraethylene glycol self-assembled monolayers. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[106]  J. Ying,et al.  Recyclable hydrophilic-hydrophobic micropatterns on glass for microarray applications. , 2007, Langmuir : the ACS journal of surfaces and colloids.

[107]  Donald Wlodkowic,et al.  Microfluidic single-cell array cytometry for the analysis of tumor apoptosis. , 2009, Analytical chemistry.

[108]  A. Jayaraman,et al.  A programmable microfluidic cell array for combinatorial drug screening. , 2012, Lab on a chip.

[109]  R D Sochol,et al.  3D printed microfluidic circuitry via multijet-based additive manufacturing. , 2016, Lab on a chip.

[110]  K. Cheung,et al.  Droplet-based microfluidic system for multicellular tumor spheroid formation and anticancer drug testing. , 2010, Lab on a chip.

[111]  Bifeng Liu,et al.  Agarose-based microwell array chip for high-throughput screening of functional microorganisms. , 2019, Talanta.

[112]  D. Wong,et al.  Genome-wide characterisation and expression profile of the grapevine ATL ubiquitin ligase family reveal biotic and abiotic stress-responsive and development-related members , 2016, Scientific Reports.

[113]  Jessica Melin,et al.  Microfluidic large-scale integration: the evolution of design rules for biological automation. , 2007, Annual review of biophysics and biomolecular structure.

[114]  Jun Liu,et al.  Analysis of nonadherent apoptotic cells by a quantum dots probe in a microfluidic device for drug screening. , 2009, Analytical chemistry.

[115]  Jun-Jie Zhu,et al.  Lab-on-a-Chip for anticancer drug screening using quantum dots probe based apoptosis assay. , 2013, Journal of biomedical nanotechnology.

[116]  Abiche H. Dewilde,et al.  A living cell quartz crystal microbalance biosensor for continuous monitoring of cytotoxic responses of macrophages to single-walled carbon nanotubes , 2011, Particle and Fibre Toxicology.

[117]  Jesse D. Gelles,et al.  Robust high-throughput kinetic analysis of apoptosis with real-time high-content live-cell imaging , 2016, Cell Death & Disease.

[118]  Jenny Zhu,et al.  Dynamic Monitoring of Cell Adhesion and Spreading on Microelectronic Sensor Arrays , 2005, Journal of biomolecular screening.

[119]  L. Basabe‐Desmonts,et al.  Microfluidic chip with pillar arrays for controlled production and observation of lipid membrane nanotubes. , 2020, Lab on a chip.

[120]  Reinhard Hickel,et al.  Real-time xCELLigence impedance analysis of the cytotoxicity of dental composite components on human gingival fibroblasts. , 2010, Dental materials : official publication of the Academy of Dental Materials.

[121]  Jian Xu,et al.  Real-time characterization of cytotoxicity using single-cell impedance monitoring. , 2012, The Analyst.

[122]  Aigars Piruska,et al.  The autofluorescence of plastic materials and chips measured under laser irradiation. , 2005, Lab on a chip.

[123]  Todd Thorsen,et al.  High-density microfluidic arrays for cell cytotoxicity analysis. , 2007, Lab on a chip.

[124]  Detlev Belder,et al.  Chip electrophoresis with mass spectrometric detection in record speed. , 2010, Lab on a chip.