Microfluidic mechanoporation for cellular delivery and analysis

Abstract

Highly efficient intracellular delivery strategies are essential for developing therapeutic, diagnostic, biological, and various biomedical applications. The recent advancement of micro/nanotechnology has focused numerous researches towards developing microfluidic device-based strategies due to the associated high throughput delivery, cost-effectiveness, robustness, and biocompatible nature. The delivery strategies can be carrier-mediated or membrane disruption-based, where membrane disruption methods find popularity due to reduced toxicity, enhanced delivery efficiency, and cell viability. Among all of the membrane disruption techniques, the mechanoporation strategies are advantageous because of no external energy source required for membrane deformation, thereby achieving high delivery efficiencies and increased cell viability into different cell types with negligible toxicity. The past two decades have consequently seen a tremendous boost in mechanoporation-based research for intracellular delivery and cellular analysis. This article provides a brief review of the most recent developments on microfluidic-based mechanoporation strategies such as microinjection, nanoneedle arrays, cell-squeezing, and hydroporation techniques with their working principle, device fabrication, cellular delivery, and analysis. Moreover, a brief discussion of the different mechanoporation strategies integrated with other delivery methods has also been provided. Finally, the advantages, limitations, and future prospects of this technique are discussed compared to other intracellular delivery techniques.

Regeneration of non-chimeric plants from DNA-free edited grapevine protoplasts

“…Possibly, undesired cell fusion could be avoided by reducing either PEG concentration or the duration of PEG treatment or by alternative methods, such as electroporation or mechanoporation ( Sharei et al., 2013 ; Chakrabarty et al., 2022 ).…”

Delivering the CRISPR/Cas9 system for engineering gene therapies: Recent cargo and delivery approaches for clinical translation

“…By understanding the relationships behind the factors that affect membrane repair mechanisms, poration methods may be improved to retain high transfection while increasing cell viability. Various studies also encounter device clogging, inverse proportionality between cell viability and transfection efficiency over time, as well as the inability to deliver nucleic acids ( Chakrabarty et al, 2021 ). Additionally, this method of delivery is optimized for in vitro work and cannot be easily adapted for in vivo applications.…”

“…Presently, this method is restricted to use with small animal models due to the large starting injection volume necessary (∼10% body weight of the mouse). As such, it is currently not appropriate for human applications, although research is ongoing to optimize this technique for larger animal use ( Chakrabarty et al, 2021 ). Deng et al (2018) investigated delivery of various cargos such as protein, siRNA, CRISPR/Cas9, plasmid DNA, and DNA nanomaterials to different cell types using a hydrodynamic delivery platform termed inertial microfluidic cell hydroporator (iMCH) ( Deng et al, 2018 ; Kang et al, 2020 ).…”

Response Surface Methodology to Efficiently Optimize Intracellular Delivery by Photoporation

“…Here, the cell membrane is transiently perturbed by physical phenomena (e.g., of mechanical, optical, thermal, magnetic and/or electrical nature), creating membrane pores through which molecules of interest can enter the cells [ 2 , 8 ]. Examples of physical membrane disruption techniques are electroporation [ 9 ], sonoporation [ 10 ], magnetofection [ 2 , 11 ] and microfluidic mechanoporation [ 12 ].…”