Microfluidic technologies for the synthesis and manipulation of biomimetic membranous nano-assemblies.

Microfluidics has been proposed as an attractive alternative to conventional bulk methods used in the generation of self-assembled biomimetic structures, particularly where there is a desire for more scalable production. The approach also allows for greater control over the self-assembly process, and parameters such as particle architecture, size, and composition can be finely tuned. Microfluidic techniques used in the generation of microscale assemblies (giant vesicles and higher-order multi-compartment assemblies) are fairly well established. These tend to rely on microdroplet templation, and the resulting structures have found use as comparmentalised motifs in artificial cells. Challenges in generating sub-micron droplets have meant that reconfiguring this approach to form nano-scale structures is not straightforward. This is beginning to change however, and recent technological advances have instigated the manufacture and manipulation of an increasingly diverse repertoire of biomimetic nano-assemblies, including liposomes, polymersomes, hybrid particles, multi-lamellar structures, cubosomes, hexosomes, nanodiscs, and virus-like particles. The following review will discuss these higher-order self-assembled nanostructures, including their biochemical and industrial applications, and techniques used in their production and analysis. We suggest ways in which existing technologies could be repurposed for the enhanced design, manufacture, and exploitation of these structures and discuss potential challenges and future research directions. By compiling recent advances in this area, it is hoped we will inspire future efforts toward establishing scalable microfluidic platforms for the generation of biomimetic nanoparticles of enhanced architectural and functional complexity.

[1]  James P. Allen Design of energy-transducing artificial cells , 2017, Proceedings of the National Academy of Sciences.

[2]  P. Ma,et al.  Cubic and Hexagonal Liquid Crystals as Drug Delivery Systems , 2014, BioMed research international.

[3]  T. Rades,et al.  Cubosomes containing the adjuvants imiquimod and monophosphoryl lipid A stimulate robust cellular and humoral immune responses. , 2013, Journal of controlled release : official journal of the Controlled Release Society.

[4]  Bastiaan C. Buddingh’,et al.  Intercellular communication between artificial cells by allosteric amplification of a molecular signal , 2020, Nature Communications.

[5]  L. Vaca,et al.  Interaction Between Virus-Like Particles (VLPs) and Pattern Recognition Receptors (PRRs) From Dendritic Cells (DCs): Toward Better Engineering of VLPs , 2020, Frontiers in Immunology.

[6]  Rui Gan,et al.  Cell-free protein synthesis: applications come of age. , 2012, Biotechnology advances.

[7]  A. Angelova,et al.  Advances in the Design of pH-Sensitive Cubosome Liquid Crystalline Nanocarriers for Drug Delivery Applications , 2020, Nanomaterials.

[8]  L. Ming,et al.  Evaluation of Extrusion Technique for Nanosizing Liposomes , 2016, Pharmaceutics.

[9]  M. Borgnia,et al.  Polymer Nanodiscs: Discoidal Amphiphilic Block Copolymer Membranes as a New Platform for Membrane Proteins , 2017, Scientific Reports.

[10]  Nikolaj Gadegaard,et al.  30 years of microfluidics , 2019, Micro and Nano Engineering.

[11]  Jan C M van Hest,et al.  Stimuli-responsive polymersomes and nanoreactors. , 2016, Journal of materials chemistry. B.

[12]  L. Prodi,et al.  Cancer-cell-targeted theranostic cubosomes. , 2014, Langmuir : the ACS journal of surfaces and colloids.

[13]  Xin Zhao,et al.  Microfluidic Generation of Nanomaterials for Biomedical Applications. , 2020, Small.

[14]  E. Cho,et al.  A Simple Evaporation Method for Large-Scale Production of Liquid Crystalline Lipid Nanoparticles with Various Internal Structures. , 2015, ACS applied materials & interfaces.

[15]  C. Drummond,et al.  Nature‐Inspired Design and Application of Lipidic Lyotropic Liquid Crystals , 2019, Advanced materials.

[16]  H. Garg,et al.  Virus Like Particles (VLP) as multivalent vaccine candidate against Chikungunya, Japanese Encephalitis, Yellow Fever and Zika Virus , 2020, Scientific Reports.

[17]  Lewis D. Blackman,et al.  Permeable Protein-Loaded Polymersome Cascade Nanoreactors by Polymerization-Induced Self-Assembly , 2017, ACS macro letters.

[18]  Frank Sainsbury,et al.  Protein cages and virus-like particles: from fundamental insight to biomimetic therapeutics. , 2020, Biomaterials science.

[19]  A. Mark,et al.  Molecular view of hexagonal phase formation in phospholipid membranes. , 2004, Biophysical journal.

[20]  N. Nguyen,et al.  Recent Advances and Future Perspectives on Microfluidic Liquid Handling , 2017, Micromachines.

[21]  C. Badger,et al.  Development of a bead-based immunoassay using virus-like particles for detection of alphaviral humoral response. , 2019, Journal of virological methods.

[22]  E. Reimhult,et al.  Triggered Release from Thermoresponsive Polymersomes with Superparamagnetic Membranes , 2016, Materials.

[23]  Meiwan Chen,et al.  Optimization of the preparation process for an oral phytantriol-based amphotericin B cubosomes , 2011 .

[24]  W. Roos,et al.  Multilamellar nanovesicles show distinct mechanical properties depending on their degree of lamellarity. , 2018, Nanoscale.

[25]  Dirk Dietrich,et al.  Nanotechnology as a Platform for the Development of Injectable Parenteral Formulations: A Comprehensive Review of the Know-Hows and State of the Art , 2020, Pharmaceutics.

[26]  G. Wagner,et al.  Large Nanodiscs: A Potential Game Changer in Structural Biology of Membrane Protein Complexes and Virus Entry , 2020, Frontiers in Bioengineering and Biotechnology.

[27]  A. Liu,et al.  Advances in Planar Lipid Bilayers and Liposomes , 2011 .

[28]  Hayley K Charlton Hume,et al.  Synthetic biology for bioengineering virus‐like particle vaccines , 2018, Biotechnology and bioengineering.

[29]  Olivier Sandre,et al.  Hybrid polymer/lipid vesicles: state of the art and future perspectives , 2013, Materials Today.

[30]  J. Chroboczek,et al.  Virus-like particles as drug delivery vectors. , 2016, Acta biochimica Polonica.

[31]  W. Meier,et al.  Vesicles in Multiple Shapes: Fine-Tuning Polymersomes’ Shape and Stability by Setting Membrane Hydrophobicity , 2017, Polymers.

[32]  J. Rumfeldt,et al.  Sonication of proteins causes formation of aggregates that resemble amyloid , 2004, Protein science : a publication of the Protein Society.

[33]  Martin Andersson,et al.  Lipid-Based Liquid Crystals As Carriers for Antimicrobial Peptides: Phase Behavior and Antimicrobial Effect. , 2016, Langmuir : the ACS journal of surfaces and colloids.

[34]  Ilia Platzman,et al.  Mastering Complexity: Towards Bottom-up Construction of Multifunctional Eukaryotic Synthetic Cells , 2018, Trends in biotechnology.

[35]  R. Mezzenga,et al.  Oil and drug control the release rate from lyotropic liquid crystals. , 2015, Journal of controlled release : official journal of the Controlled Release Society.

[36]  Alfonso M. Gañán-Calvo,et al.  Focusing capillary jets close to the continuum limit , 2007 .

[37]  V. Labhasetwar,et al.  Biophysical interactions with model lipid membranes: applications in drug discovery and drug delivery. , 2009, Molecular pharmaceutics.

[38]  R. Mezzenga,et al.  Controlling enzymatic activity and kinetics in swollen mesophases by physical nano-confinement. , 2014, Nanoscale.

[39]  K. Castiglione,et al.  Polymersomes for biotechnological applications: Large‐scale production of nano‐scale vesicles , 2017, Engineering in life sciences.

[40]  Oscar Ces,et al.  Artificial cell mimics as simplified models for the study of cell biology , 2017, Experimental biology and medicine.

[41]  Petra Schwille,et al.  Liposomes and polymersomes: a comparative review towards cell mimicking. , 2018, Chemical Society reviews.

[42]  O. Ces,et al.  Microfluidic generation of encapsulated droplet interface bilayer networks (multisomes) and their use as cell-like reactors. , 2016, Chemical communications.

[43]  Olivier Sandre,et al.  Polymersome shape transformation at the nanoscale. , 2013, ACS nano.

[44]  P. Alves,et al.  Virus-like particles in vaccine development , 2010, Expert review of vaccines.

[45]  J. V. Hest,et al.  Methods for production of uniform small-sized polymersome with rigid membrane , 2016 .

[46]  Extraction and reconstitution of membrane proteins into lipid nanodiscs encased by zwitterionic styrene-maleic amide copolymers , 2020, Scientific Reports.

[47]  I. Foulds,et al.  Three-dimensional parallelization of microfluidic droplet generators for a litre per hour volume production of single emulsions. , 2014, Lab on a chip.

[48]  O. Vinogradova,et al.  Nanodiscs and solution NMR: preparation, application and challenges , 2017, Nanotechnology reviews.

[49]  Soodabeh Davaran,et al.  Liposome: classification, preparation, and applications , 2013, Nanoscale Research Letters.

[50]  Bastian E. Rapp,et al.  Let there be chip—towards rapid prototyping of microfluidic devices: one-step manufacturing processes , 2011 .

[51]  M. A. Roberts,et al.  Edinburgh Research Explorer Synthetic biology: biology by design , 2014 .

[52]  R. Langer,et al.  mRNA vaccine delivery using lipid nanoparticles. , 2016, Therapeutic delivery.

[53]  S. Sligar,et al.  Nanodiscs for structural and functional studies of membrane proteins , 2016, Nature Structural &Molecular Biology.

[54]  S. Gui,et al.  Factors affecting the structure of lyotropic liquid crystals and the correlation between structure and drug diffusion , 2018, RSC advances.

[55]  Juan Pérez-Mercader,et al.  Microfluidic fabrication of polymersomes enclosing an active Belousov-Zhabotinsky (BZ) reaction: Effect on their stability of solute concentrations in the external media. , 2016, Colloids and surfaces. B, Biointerfaces.

[56]  H. Petry,et al.  The use of virus-like particles for gene transfer. , 2003, Current opinion in molecular therapeutics.

[57]  C. Drummond,et al.  Amphiphilic brush polymers produced using the RAFT polymerisation method stabilise and reduce the cell cytotoxicity of lipid lyotropic liquid crystalline nanoparticles. , 2016, Faraday discussions.

[58]  J. Jouhet Importance of the hexagonal lipid phase in biological membrane organization , 2013, Front. Plant Sci..

[59]  P. Štěpánek,et al.  Microfluidic-Assisted Engineering of Quasi-Monodisperse pH-Responsive Polymersomes toward Advanced Platforms for the Intracellular Delivery of Hydrophilic Therapeutics. , 2019, Langmuir : the ACS journal of surfaces and colloids.

[60]  Junyoung Kim,et al.  Polymersome-based modular nanoreactors with size-selective transmembrane permeability. , 2020, ACS applied materials & interfaces.

[61]  Guoliang Zhen,et al.  Salt induced lamellar to bicontinuous cubic phase transitions in cationic nanoparticles. , 2012, The journal of physical chemistry. B.

[62]  C. Drummond,et al.  Design of ultra-swollen lipidic mesophases for the crystallization of membrane proteins with large extracellular domains , 2018, Nature Communications.

[63]  Peter B Howell,et al.  Toolbox for the design of optimized microfluidic components. , 2006, Lab on a chip.

[64]  Peter Beike,et al.  Intermolecular And Surface Forces , 2016 .

[65]  T. Oka,et al.  Preparation and Characterization of SN-38-Encapsulated Phytantriol Cubosomes Containing α-Monoglyceride Additives. , 2016, Chemical & pharmaceutical bulletin.

[66]  Nathan S. Mosier,et al.  Nanoscale Drug Delivery Systems: From Medicine to Agriculture , 2020, Frontiers in Bioengineering and Biotechnology.

[67]  P. Wright,et al.  Polymersome production on a microfluidic platform using pH sensitive block copolymers. , 2010, Lab on a chip.

[68]  A. Zeltiņš,et al.  Construction and Characterization of Virus-Like Particles: A Review , 2012, Molecular Biotechnology.

[69]  D. DeVoe,et al.  Microfluidic remote loading for rapid single-step liposomal drug preparation. , 2014, Lab on a chip.

[70]  Yi Yan Yang,et al.  Engineering Polymersomes for Diagnostics and Therapy , 2018, Advanced healthcare materials.

[71]  S. Sligar,et al.  Microfluidic platform for efficient Nanodisc assembly, membrane protein incorporation, and purification. , 2017, Lab on a chip.

[72]  D. Weitz,et al.  Photothermal-responsive nanosized hybrid polymersome as versatile therapeutics codelivery nanovehicle for effective tumor suppression , 2019, Proceedings of the National Academy of Sciences.

[73]  X. Mulet,et al.  Controlling nanostructure and lattice parameter of the inverse bicontinuous cubic phases in functionalised phytantriol dispersions. , 2013, Journal of colloid and interface science.

[74]  S. Gurunathan,et al.  The promise of mRNA vaccines: a biotech and industrial perspective , 2020, npj Vaccines.

[75]  D. Rentsch,et al.  Wavelength-Selective Light-Responsive DASA-Functionalized Polymersome Nanoreactors. , 2018, Journal of the American Chemical Society.

[76]  S. Gurunathan,et al.  The promise of mRNA vaccines: a biotech and industrial perspective , 2020, npj Vaccines.

[77]  O. Ces,et al.  Engineering swollen cubosomes using cholesterol and anionic lipids. , 2019, Langmuir : the ACS journal of surfaces and colloids.

[78]  R. Templer,et al.  Inverse lyotropic phases of lipids and membrane curvature , 2006, Journal of physics. Condensed matter : an Institute of Physics journal.

[79]  A. Yaghmur,et al.  A hydrodynamic flow focusing microfluidic device for the continuous production of hexosomes based on docosahexaenoic acid monoglyceride. , 2019, Physical chemistry chemical physics : PCCP.

[80]  S. Matosevic,et al.  Layer-by-layer Cell Membrane Assembly , 2013, Nature chemistry.

[81]  Stephan Herminghaus,et al.  Vesicles-on-a-chip: A universal microfluidic platform for the assembly of liposomes and polymersomes , 2016, The European physical journal. E, Soft matter.

[82]  M. Andersson,et al.  Peptide-Loaded Cubosomes Functioning as an Antimicrobial Unit against Escherichia coli. , 2019, ACS applied materials & interfaces.

[83]  A. Salehi-Reyhani,et al.  Droplet microfluidics for the construction of compartmentalised model membranes. , 2018, Lab on a chip.

[84]  Oscar Ces,et al.  Microfluidics for Artificial Life: Techniques for Bottom-Up Synthetic Biology , 2019, Micromachines.

[85]  Susan Hua,et al.  Advances and Challenges of Liposome Assisted Drug Delivery , 2015, Front. Pharmacol..

[86]  S. Moghimi,et al.  Cubosomes and hexosomes as versatile platforms for drug delivery. , 2015, Therapeutic delivery.

[87]  Jing Qiao,et al.  Natural supramolecular building blocks: from virus coat proteins to viral nanoparticles. , 2012, Chemical Society reviews.

[88]  O. Ces,et al.  Programming membrane permeability using integrated membrane pores and blockers as molecular regulators. , 2017, Chemical communications.

[89]  T. Nisisako,et al.  Microfluidic large-scale integration on a chip for mass production of monodisperse droplets and particles. , 2008, Lab on a chip.

[90]  L. Prodi,et al.  Drug-loaded fluorescent cubosomes: versatile nanoparticles for potential theranostic applications. , 2013, Langmuir : the ACS journal of surfaces and colloids.

[91]  S. Sligar,et al.  Nanodiscs in Membrane Biochemistry and Biophysics. , 2017, Chemical reviews.

[92]  E. Wachtel,et al.  Hexosome and hexagonal phases mediated by hydration and polymeric stabilizer. , 2007, Langmuir : the ACS journal of surfaces and colloids.

[93]  Chang Liu,et al.  Microfluidic patterning of nanodisc lipid bilayers and multiplexed analysis of protein interaction. , 2008, Lab on a chip.

[94]  Juan J de Pablo,et al.  Stimuli-Responsive Cubosomes Formed from Blue Phase Liquid Crystals. , 2015, Advanced materials.

[95]  M. Stevens,et al.  Cubosomes: The Next Generation of Smart Lipid Nanoparticles? , 2019, Angewandte Chemie.

[96]  D. Lasič,et al.  The mechanism of vesicle formation. , 1988, The Biochemical journal.

[97]  J. Dennis,et al.  The lipid composition of autophagic vacuoles regulates expression of multilamellar bodies , 2005, Journal of Cell Science.

[98]  S. N. Lai,et al.  Artificial Cells Capable of Long-Lived Protein Synthesis by Using Aptamer Grafted Polymer Hydrogel. , 2019, ACS synthetic biology.

[99]  Aghiad Ghazal,et al.  Microfluidic Platform for the Continuous Production and Characterization of Multilamellar Vesicles: A Synchrotron Small-Angle X-ray Scattering (SAXS) Study. , 2017, The journal of physical chemistry letters.

[100]  A Paul Alivisatos,et al.  High-temperature microfluidic synthesis of CdSe nanocrystals in nanoliter droplets. , 2005, Journal of the American Chemical Society.

[101]  D. Weitz,et al.  Geometrically mediated breakup of drops in microfluidic devices. , 2003, Physical review letters.

[102]  Vivek S. Dave,et al.  Nanostructured Cubosomes in a Thermoresponsive Depot System: An Alternative Approach for the Controlled Delivery of Docetaxel , 2015, AAPS PharmSciTech.

[103]  U. Raviv,et al.  Studying viruses using solution X-ray scattering , 2020, Biophysical Reviews.

[104]  Andreas Wagner,et al.  Liposome Technology for Industrial Purposes , 2010, Journal of drug delivery.

[105]  R. Olsthoorn,et al.  Drug Delivery via Cell Membrane Fusion Using Lipopeptide Modified Liposomes , 2016, ACS Central Science.

[106]  S. Charette,et al.  Lipid Composition of Multilamellar Bodies Secreted by Dictyostelium discoideum Reveals Their Amoebal Origin , 2013, Eukaryotic Cell.

[107]  J. Spatz,et al.  Bottom-Up Assembly of Functional Intracellular Synthetic Organelles by Droplet-Based Microfluidics. , 2020, Small.

[108]  Yuval Elani,et al.  Interfacing Living and Synthetic Cells as an Emerging Frontier in Synthetic Biology , 2020, Angewandte Chemie.

[109]  P. Yalavarthi,et al.  Cubosomes as targeted drug delivery systems - a biopharmaceutical approach. , 2014, Current drug discovery technologies.

[110]  Eleanor Stride,et al.  Liposome production by microfluidics: potential and limiting factors , 2016, Scientific Reports.

[111]  A. Angelova,et al.  pH-Responsiveness of Hexosomes and Cubosomes for Combined Delivery of Brucea Javanica Oil and Doxorubicin. , 2019, Langmuir : the ACS journal of surfaces and colloids.

[112]  Dirk van Swaay,et al.  Microfluidic methods for forming liposomes. , 2013, Lab on a chip.

[113]  Matthias Schulz,et al.  Mixed Hybrid Lipid/Polymer Vesicles as a Novel Membrane Platform. , 2015, Macromolecular rapid communications.

[114]  I. Alves,et al.  Microfluidic diffusional sizing probes lipid nanodiscs formation. , 2020, Biochimica et biophysica acta. Biomembranes.

[115]  J. Bouwstra,et al.  Small angle X-ray scattering: possibilities and limitations in characterization of vesicles. , 1993, Chemistry and physics of lipids.

[116]  O. Ces,et al.  Functionalizing cell-mimetic giant vesicles with encapsulated bacterial biosensors , 2018, Interface Focus.

[117]  Aleksandr Ovsianikov,et al.  Functional 3D Printing for Microfluidic Chips , 2019, Advanced Materials Technologies.

[118]  W. Wilson,et al.  Dual pH-sensitive liposomes with low pH-triggered sheddable PEG for enhanced tumor-targeted drug delivery. , 2019, Nanomedicine.

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

[120]  C. Batt,et al.  Analysis of a laminar-flow diffusional mixer for directed self-assembly of liposomes. , 2012, Biomicrofluidics.

[121]  G. Battaglia,et al.  Chemotactic synthetic vesicles: Design and applications in blood-brain barrier crossing , 2016, Science Advances.

[122]  Mansoor Nasir,et al.  Hydrodynamic focusing—a versatile tool , 2011, Analytical and Bioanalytical Chemistry.

[123]  T. Heimburg,et al.  Thermodynamics of lipid multi-lamellar vesicles in presence of sterols at high hydrostatic pressure , 2017, Scientific Reports.

[124]  Jaeuk Sung,et al.  Microfluidics Synthesis of Gene Silencing Cubosomes. , 2018, ACS nano.

[125]  W. Hur,et al.  Liver-specific Gene Delivery Using Engineered Virus-Like Particles of Hepatitis E Virus , 2019, Scientific Reports.

[126]  Don L DeVoe,et al.  High-Throughput Continuous Flow Production of Nanoscale Liposomes by Microfluidic Vertical Flow Focusing. , 2015, Small.

[127]  S. Haam,et al.  Dengue Virus-Polymersome Hybrid Nanovesicles for Advanced Drug Screening using Real-Time Single Nanoparticle-Virus Tracking. , 2020, ACS applied materials & interfaces.

[128]  Yanhui Zhao,et al.  Microfluidic Hydrodynamic Focusing for Synthesis of Nanomaterials. , 2016, Nano today.

[129]  Sepp D. Kohlwein,et al.  Cubic membranes: a legend beyond the Flatland* of cell membrane organization , 2006, The Journal of cell biology.

[130]  Naser Karimi,et al.  Application of Various Types of Liposomes in Drug Delivery Systems , 2017, Advanced pharmaceutical bulletin.

[131]  Reza Ghodssi,et al.  Capillary Microfluidics-Assembled Virus-like Particle Bionanoreceptor Interfaces for Label-Free Biosensing. , 2017, ACS applied materials & interfaces.

[132]  Shoji Takeuchi,et al.  Artificial Cell Membrane Systems for Biosensing Applications. , 2017, Analytical chemistry.

[133]  A. Meijering,et al.  Octanol-assisted liposome assembly on chip , 2016, Nature Communications.

[134]  Maaruthy Yelleswarapu,et al.  Microfluidic Assembly of Monodisperse Vesosomes as Artificial Cell Models. , 2017, Journal of the American Chemical Society.

[135]  M. Bachmann,et al.  Major findings and recent advances in virus-like particle (VLP)-based vaccines. , 2017, Seminars in immunology.