Flow-based assembly of nucleic acid-loaded polymer nanoparticles
Zeyan Xu A , Joshua McCarrol B C D and Martina H. Stenzel
A D *
A School of Chemistry, University of New South Wales, Sydney, NSW 2052, Australia.
B Children’s Cancer Institute, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW 2052, Australia.
C School of Women’s and Children’s Health, University of New South Wales, Sydney, NSW 2052, Australia.
D UNSW RNA Institute, University of New South Wales, Sydney, NSW 2052, Australia.
Abstract
Since the development of messenger RNA (mRNA)-based SARS-CoV-2 (COVID-19) vaccines, there is increased public awareness of the importance of nanoparticles, in this case lipid nanoparticles, to ensure safe delivery of an active compound. To ensure the formation of high-quality nanoparticles with reproducible results, these lipid nanoparticles are assembled with the nucleic acid drug using flow-based devices. Although flow assembly using lipid nanoparticles for nucleic acid delivery is well described in the literature, only a few examples use polymers. This is surprising because the field of polymers for nucleic acid delivery is substantial as hundreds of polymers for nucleic acid delivery have been reported in the literature. In this review, we discuss several aspects of flow-based assembly of nucleic acid-loaded polymer nanoparticles. Initially, we introduce the concept of chip-based or capillary-based systems that can be either used as single-phase or multiphase systems. Initially, researchers have to choose the type of mixing, which can be active or passive. The type of flow, laminar or turbulent, also significantly affects the quality of the nanoparticles. We then present the type of polymers that have so far been assembled with mRNA, small interfering RNA (siRNA) or plasmid DNA (pDNA) using flow devices. We discuss effects such as flow rate, concentration and polymer lengths on the outcome. To conclude, we highlight how flow assembly is an excellent way to generate well-defined nanoparticles including polyplexes in a reproducible manner.
Keywords: DNA, drug delivery, flow assembly, gene therapy, microfluidics, nanomedicine, polymers, RNA, self‐assembly.
References
1 Mitchell MJ, Billingsley MM, Haley RM, Wechsler ME, Peppas NA, Langer R. Engineering precision nanoparticles for drug delivery. Nat Rev Drug Discov 2021; 20: 101-124.
| Crossref | Google Scholar |
2 Halwani AA. Development of pharmaceutical nanomedicines: from the bench to the market. Pharmaceutics 2022; 14: 106.
| Crossref | Google Scholar |
3 Shan X, Gong X, Li J, Wen J, Li Y, Zhang Z. Current approaches of nanomedicines in the market and various stage of clinical translation. Acta Pharm Sin B 2022; 12: 3028-3048.
| Crossref | Google Scholar |
4 Hou X, Zaks T, Langer R, Dong Y. Lipid nanoparticles for mRNA delivery. Nat Rev Mater 2021; 6: 1078-1094.
| Crossref | Google Scholar |
5 Kumar R, Santa Chalarca CF, Bockman MR, Bruggen CV, Grimme CJ, Dalal RJ, et al. Polymeric delivery of therapeutic nucleic acids. Chem Rev 2021; 121: 11527-11652.
| Crossref | Google Scholar |
6 Paunovska K, Loughrey D, Dahlman JE. Drug delivery systems for RNA therapeutics. Nat Rev Genet 2022; 23: 265-280.
| Crossref | Google Scholar |
7 Zhang L, Nguyen TLU, Bernard J, Davis TP, Barner-Kowollik C, Stenzel MH. Shell-cross-linked micelles containing cationic polymers synthesized via the RAFT process: toward a more biocompatible gene delivery system. Biomacromolecules 2007; 8: 2890-2901.
| Crossref | Google Scholar |
8 Hui Y, Yi X, Hou F, Wibowo D, Zhang F, Zhao D, et al. Role of nanoparticle mechanical properties in cancer drug delivery. ACS Nano 2019; 13: 7410-7424.
| Crossref | Google Scholar |
9 Donahue ND, Acar H, Wilhelm S. Concepts of nanoparticle cellular uptake, intracellular trafficking, and kinetics in nanomedicine. Adv Drug Delivery Rev 2019; 143: 68-96.
| Crossref | Google Scholar |
10 Stenzel MH. The trojan horse goes wild: the effect of drug loading on the behavior of nanoparticles. Angew Chem Int Ed Engl 2021; 60: 2202-2206.
| Crossref | Google Scholar |
11 Hoshyar N, Gray S, Han H, Bao G. The effect of nanoparticle size on in vivo pharmacokinetics and cellular interaction. Nanomed 2016; 11: 673-692.
| Crossref | Google Scholar |
12 Choi CHJ, Zuckerman JE, Webster P, Davis ME. Targeting kidney mesangium by nanoparticles of defined size. Proc Natl Acad Sci USA 2011; 108: 6656-6661.
| Crossref | Google Scholar |
13 Perrault SD, Walkey C, Jennings T, Fischer HC, Chan WCW. Mediating tumor targeting efficiency of nanoparticles through design. Nano Lett 2009; 9: 1909-1915.
| Crossref | Google Scholar |
14 Soo Choi H, Liu W, Misra P, Tanaka E, Zimmer JP, Itty Ipe B, et al. Renal clearance of quantum dots. Nat Biotechnol 2007; 25: 1165-1170.
| Crossref | Google Scholar |
15 Gustafson HH, Holt-Casper D, Grainger DW, Ghandehari H. Nanoparticle uptake: the phagocyte problem. Nano Today 2015; 10: 487-510.
| Crossref | Google Scholar |
16 Alexis F, Pridgen E, Molnar LK, Farokhzad OC. Factors affecting the clearance and biodistribution of polymeric nanoparticles. Mol Pharmaceutics 2008; 5: 505-515.
| Crossref | Google Scholar |
17 Blanco E, Shen H, Ferrari M. Principles of nanoparticle design for overcoming biological barriers to drug delivery. Nat Biotechnol 2015; 33: 941-951.
| Crossref | Google Scholar |
18 Cabral H, Matsumoto Y, Mizuno K, Chen Q, Murakami M, Kimura M, et al. Accumulation of sub-100 nm polymeric micelles in poorly permeable tumours depends on size. Nat Nanotechnol 2011; 6: 815-823.
| Crossref | Google Scholar |
19 Yu W, Liu R, Zhou Y, Gao H. Size-tunable strategies for a tumor targeted drug delivery system. ACS Cent Sci 2020; 6: 100-116.
| Crossref | Google Scholar |
20 Ulkoski D, Scholz C. Impact of cationic charge density and pegylated poly(amino acid) tercopolymer architecture on their use as gene delivery vehicles. Part 2: DNA protection, stability, cytotoxicity, and transfection efficiency. Macromol Biosci 2018; 18: 1800109.
| Crossref | Google Scholar |
21 Bauer M, Tauhardt L, Lambermont-Thijs HML, Kempe K, Hoogenboom R, Schubert US, et al. Rethinking the impact of the protonable amine density on cationic polymers for gene delivery: a comparative study of partially hydrolyzed poly(2‐ethyl‐2‐oxazoline)s and linear poly(ethylene imine)s. Eur J Pharm Biopharm 2018; 133: 112-121.
| Crossref | Google Scholar |
22 Jiang Y, Stenzel M. Drug delivery vehicles based on albumin-polymer conjugates. Macromol Biosci 2016; 16: 791-802.
| Crossref | Google Scholar |
23 Joshi N, Liu DL, Dickson KA, Marsh DJ, Ford CE, Stenzel MH. An organotypic model of high-grade serous ovarian cancer to test the anti-metastatic potential of ROR2 targeted polyion complex nanoparticles. J Mater Chem B 2021; 9: 9123-9135.
| Crossref | Google Scholar |
24 Jiang Y, Wong CK, Stenzel MH. An oligonucleotide transfection vector based on hsa and pdmaema conjugation: effect of polymer molecular weight on cell proliferation and on multicellular tumor spheroids. Macromol Biosci 2015; 15: 965-978.
| Crossref | Google Scholar |
25 Jiang Y, Lu H, Khine YY, Dag A, Stenzel MH. Polyion complex micelle based on albumin-polymer conjugates: multifunctional oligonucleotide transfection vectors for anticancer chemotherapeutics. Biomacromolecules 2014; 15: 4195-4205.
| Crossref | Google Scholar |
26 Đorđević S, Gonzalez MM, Conejos-Sánchez I, Carreira B, Pozzi S, Acúrcio RC, et al. Current hurdles to the translation of nanomedicines from bench to the clinic. Deliv and Transl Res 2022; 12: 500-525.
| Crossref | Google Scholar |
27 Hirota S, de Ilarduya CT, Barron LG, Szoka Jr FC. Simple mixing device to reproducibly prepare cationic lipid–DNA complexes (lipoplexes). Biotechniques 1999; 27: 286-290.
| Google Scholar |
28 Maeki M, Kimura N, Sato Y, Harashima H, Tokeshi M. Advances in microfluidics for lipid nanoparticles and extracellular vesicles and applications in drug delivery systems. Adv Drug Deliv Rev 2018; 128: 84-100.
| Crossref | Google Scholar |
29 Jung HN, Lee SY, Lee S, Youn H, Im HJ. Lipid nanoparticles for delivery of rna therapeutics: current status and the role of in vivo imaging. Theranostics 2022; 12: 7509-7531.
| Crossref | Google Scholar |
31 Johnson BK, Prud’homme RK. Flash nanoprecipitation of organic actives and block copolymers using a confined impinging jets mixer. Aust J Chem 2003; 56: 1021-1024.
| Crossref | Google Scholar |
32 Feng J, Markwalter CE, Tian C, Armstrong M, Prud’homme RK. Translational formulation of nanoparticle therapeutics from laboratory discovery to clinical scale. J Transl Med 2019; 17: 200.
| Crossref | Google Scholar |
33 Zhang H, Zhu Y, Shen Y. Microfluidics for cancer nanomedicine: from fabrication to evaluation. Small 2018; 14: 1800360.
| Crossref | Google Scholar |
34 Lee CY, Chang CL, Wang YN, Fu LM. Microfluidic mixing: a review. Int J Mol Sci 2011; 12: 3263-3287.
| Crossref | Google Scholar |
35 Ober TJ, Foresti D, Lewis JA. Active mixing of complex fluids at the microscale. Proc Natl Acad Sci USA 2015; 112: 12293-12298.
| Crossref | Google Scholar |
36 Lu M, Ozcelik A, Grigsby CL, Zhao Y, Guo F, Leong KW, et al. Microfluidic hydrodynamic focusing for synthesis of nanomaterials. Nano Today 2016; 11: 778-792.
| Crossref | Google Scholar |
37 Liu D, Zhang H, Fontana F, Hirvonen JT, Santos HA. Current developments and applications of microfluidic technology toward clinical translation of nanomedicines. Adv Drug Deliv Rev 2018; 128: 54-83.
| Crossref | Google Scholar |
38 Debus H, Beck-Broichsitter M, Kissel T. Optimized preparation of PDNA/poly(ethylene imine) polyplexes using a microfluidic system. Lab Chip 2012; 12: 2498-2506.
| Crossref | Google Scholar |
39 Stroock AD, Dertinger SKW, Ajdari A, Mezic’ I, Stone HA, Whitesides GM. Chaotic mixer for microchannels. Science 2002; 295: 647-651.
| Crossref | Google Scholar |
40 Williams MS, Longmuir KJ, Yager P. A practical guide to the staggered herringbone mixer. Lab Chip 2008; 8: 1121-1129.
| Crossref | Google Scholar |
41 Ali MS, Hooshmand N, El-Sayed M, Labouta HI. Microfluidics for development of lipid nanoparticles: paving the way for nucleic acids to the clinic. ACS Appl Bio Mater 2021;
| Crossref | Google Scholar |
42 Reynolds WC, Parekh DE, Juvet PJD, Lee MJD. Bifurcating and blooming jets. Ann Rev Fluid Mechanics 2003; 35: 295-315.
| Crossref | Google Scholar |
43 He B, Burke BJ, Zhang X, Zhang R, Regnier FE. A picoliter-volume mixer for microfluidic analytical systems. Anal Chem 2001; 73: 1942-1947.
| Crossref | Google Scholar |
44 Jen C-P, Wu C-Y, Lin Y-C, Wu C-Y. Design and simulation of the micromixer with chaotic advection in twisted microchannels. Lab Chip 2003; 3: 77-81.
| Crossref | Google Scholar |
45 Protopapa G, Bono N, Visone R, D’Alessandro F, Rasponi M, Candiani G. A new microfluidic platform for the highly reproducible preparation of non-viral gene delivery complexes. Lab Chip 2023; 23: 136-145.
| Crossref | Google Scholar |
46 Koh CG, Kang X, Xie Y, Fei Z, Guan J, Yu B, et al. Delivery of polyethylenimine/DNA complexes assembled in a microfluidics device. Mol Pharmaceutics 2009; 6: 1333-1342.
| Crossref | Google Scholar |
47 Wilson DR, Mosenia A, Suprenant MP, Upadhya R, Routkevitch D, Meyer RA, et al. Continuous microfluidic assembly of biodegradable poly(beta-amino ester)/DNA nanoparticles for enhanced gene delivery. J Biomed Mater Res Part A 2017; 105: 1813-1825.
| Crossref | Google Scholar |
48 Agnoletti M, Bohr A, Thanki K, Wan F, Zeng X, Boetker JP, et al. Inhalable siRNA-loaded nano-embedded microparticles engineered using microfluidics and spray drying. Europ J Pharm Biopharm 2017; 120: 9-21.
| Crossref | Google Scholar |
49 Zhigaltsev IV, Belliveau N, Hafez I, Leung AKK, Huft J, Hansen C, et al. Bottom–up design and synthesis of limit size lipid nanoparticle systems with aqueous and triglyceride cores using millisecond microfluidic mixing. Langmuir 2012; 28: 3633-3640.
| Crossref | Google Scholar |
50 Belliveau NM, Huft J, Lin PJC, Chen S, Leung AKK, Leaver TJ, et al. Microfluidic synthesis of highly potent limit-size lipid nanoparticles for in vivo delivery of siRNA. Mol Ther Nucleic Acids 2012; 1: e37.
| Crossref | Google Scholar |
51 Chen T, Peng Y, Qiu M, Yi C, Xu Z. Recent advances in mixing-induced nanoprecipitation: from creating complex nanostructures to emerging applications beyond biomedicine. Nanoscale 2023; 15: 3594-3609.
| Crossref | Google Scholar |
52 Karnik R, Gu F, Basto P, Cannizzaro C, Dean L, Kyei-Manu W, et al. Microfluidic platform for controlled synthesis of polymeric nanoparticles. Nano Lett 2008; 8: 2906-2912.
| Crossref | Google Scholar |
53 Liu Y, Yang G, Zou D, Hui Y, Nigam K, Middelberg APJ, et al. Formulation of nanoparticles using mixing-induced nanoprecipitation for drug delivery. Ind Eng Chem Res 2020; 59: 4134-4149.
| Crossref | Google Scholar |
54 Saad WS, Prud’homme RK. Principles of nanoparticle formation by flash nanoprecipitation. Nano Today 2016; 11: 212-227.
| Crossref | Google Scholar |
55 Yan X, Bernard J, Ganachaud F. Nanoprecipitation as a simple and straightforward process to create complex polymeric colloidal morphologies. Adv Colloid Interface Sci 2021; 294: 102474.
| Crossref | Google Scholar |
56 Ma J, Lee SM-Y, Yi C, Li C-W. Controllable synthesis of functional nanoparticles by microfluidic platforms for biomedical applications – a review. Lab Chip 2017; 17: 209-226.
| Crossref | Google Scholar |
58 Nunes JK, Tsai SSH, Wan J, Stone HA. Dripping and jetting in microfluidic multiphase flows applied to particle and fibre synthesis. J Phys D: Appl Phys 2013; 46: 114002.
| Crossref | Google Scholar |
59 Moragues T, Arguijo D, Beneyton T, Modavi C, Simutis K, Abate AR, et al. Droplet-based microfluidics. Nat Rev Methods Primers 2023; 3: 32.
| Crossref | Google Scholar |
60 Amirifar L, Besanjideh M, Nasiri R, Shamloo A, Nasrollahi F, de Barros NR, et al. Droplet-based microfluidics in biomedical applications. Biofabrication 2022; 14: 022001.
| Crossref | Google Scholar |
61 Tian F, Cai L, Liu C, Sun J. Microfluidic technologies for nanoparticle formation. Lab Chip 2022; 22: 512-529.
| Crossref | Google Scholar |
62 He F, Zhang M-J, Wang W, Cai Q-W, Su Y-Y, Liu Z, et al. Designable polymeric microparticles from droplet microfluidics for controlled drug release. Adv Mater Technol 2019; 4: 1800687.
| Crossref | Google Scholar |
63 Grigsby CL, Ho Y-P, Lin C, Engbersen JFJ, Leong KW. Microfluidic preparation of polymer-nucleic acid nanocomplexes improves nonviral gene transfer. Sci Rep 2013; 3: 3155.
| Crossref | Google Scholar |
64 Riahi R, Tamayol A, Shaegh SAM, Ghaemmaghami AM, Dokmeci MR, Khademhosseini A. Microfluidics for advanced drug delivery systems. Curr Opin Chem Eng 2015; 7: 101-112.
| Crossref | Google Scholar |
65 Deveza L, Ashoken J, Castaneda G, Tong X, Keeney M, Han L-H, et al. Microfluidic synthesis of biodegradable polyethylene-glycol microspheres for controlled delivery of proteins and DNA nanoparticles. ACS Biomater Sci Eng 2015; 1: 157-165.
| Crossref | Google Scholar |
66 Zhang L, Chen Q, Ma Y, Sun J. Microfluidic methods for fabrication and engineering of nanoparticle drug delivery systems. ACS Appl Bio Mater 2020; 3: 107-120.
| Crossref | Google Scholar |
67 Luo G, Du L, Wang Y, Lu Y, Xu J. Controllable preparation of particles with microfluidics. Particuology 2011; 9: 545-558.
| Crossref | Google Scholar |
68 Zhao C-X, He L, Qiao SZ, Middelberg APJ. Nanoparticle synthesis in microreactors. Chem Eng Sci 2011; 66: 1463-1479.
| Crossref | Google Scholar |
69 Soni G, Yadav KS. Applications of nanoparticles in treatment and diagnosis of leukemia. Mater Sci Eng C 2015; 47: 156-164.
| Crossref | Google Scholar |
70 Khan IU, Serra CA, Anton N, Vandamme TF. Production of nanoparticle drug delivery systems with microfluidics tools. Expert Opin Drug Deliv 2015; 12: 547-562.
| Crossref | Google Scholar |
71 Loy DM, Krzysztoń R, Lächelt U, Rädler JO, Wagner E. Controlling nanoparticle formulation: a low-budget prototype for the automation of a microfluidic platform. Processes 2021; 9: 129.
| Crossref | Google Scholar |
72 Tavakoli Naeini A, Soliman OY, Alameh MG, Lavertu M, Buschmann MD. Automated in-line mixing system for large scale production of chitosan-based polyplexes. J Colloid Interface Sci 2017; 500: 253-263.
| Crossref | Google Scholar |
73 Zoqlam R, Morris CJ, Akbar M, Alkilany AM, Hamdallah SI, Belton P, et al. Evaluation of the benefits of microfluidic-assisted preparation of polymeric nanoparticles for DNA delivery. Mater Sci Eng C 2021; 127: 112243.
| Crossref | Google Scholar |
74 Santhanes D, Wilkins A, Zhang H, John Aitken R, Liang M. Microfluidic formulation of lipid/polymer hybrid nanoparticles for plasmid DNA (pdna) delivery. International J Pharm 2022; 627: 122223.
| Crossref | Google Scholar |
75 Ita K. Polyplexes for gene and nucleic acid delivery: progress and bottlenecks. Eur J Pharm Sci 2020; 150: 105358.
| Crossref | Google Scholar |
76 Tros de Ilarduya C, Sun Y, Düzgüneş N. Gene delivery by lipoplexes and polyplexes. Eur J Pharm Sci 2010; 40: 159-170.
| Crossref | Google Scholar |
77 Elouahabi A, Ruysschaert J-M. Formation and intracellular trafficking of lipoplexes and polyplexes. Mol Ther 2005; 11: 336-347.
| Crossref | Google Scholar |
78 Miladi K, Sfar S, Fessi H, Elaissari A. Nanoprecipitation process: from particle preparation to in vivo applications. In: Vauthier C, Ponchel G, editors. Polymer nanoparticles for nanomedicines: a guide for their design, preparation and development. Cham, Switzerland: Springer International Publishing; 2016. pp. 17–53.
79 Zhang L, Chen Q, Ma Y, Sun J. Microfluidic methods for fabrication and engineering of nanoparticle drug delivery systems. ACS Appl Bio Mater 2019; 3: 107-120.
| Crossref | Google Scholar |
80 Brocchini S, James K, Tangpasuthadol V, Kohn J. A combinatorial approach for polymer design. J Am Chem Soc 1997; 119: 4553-4554.
| Crossref | Google Scholar |
81 Roces CB, Christensen D, Perrie Y. Translating the fabrication of protein-loaded poly (lactic-co-glycolic acid) nanoparticles from bench to scale-independent production using microfluidics. Drug Deliv Transl Res 2020; 10: 582-593.
| Crossref | Google Scholar |
82 Kastner E, Kaur R, Lowry D, Moghaddam B, Wilkinson A, Perrie Y. High-throughput manufacturing of size-tuned liposomes by a new microfluidics method using enhanced statistical tools for characterization. Int J Pharm 2014; 477: 361-368.
| Crossref | Google Scholar |
83 Wild A, Leaver T, Taylor RJ. Bifurcating mixers and methods of their use and manufacture. Google Patents; 2018. US10076730B2. Available at https://patents.google.com/patent/US10076730B2/en
85 Bono N, Ponti F, Mantovani D, Candiani G. Non-viral in vitro gene delivery: it is now time to set the bar! Pharmaceutics 2020; 12: 183.
| Crossref | Google Scholar |
86 Wu J, Fang H, Zhang J, Yan S. Modular microfluidics for life sciences. J Nanobiotechnology 2023; 21: 85.
| Crossref | Google Scholar |
