Mechanodelivery of nanoparticles to the cytoplasm of living cells

Abstract

Nanotechnology has opened up the opportunity to probe, sense, and manipulate the chemical environment of biological systems with an unprecedented level of spatiotemporal control. A major obstacle to the full realization of these novel technologies is the lack of a general, robust, and simple method for the delivery of arbitrary nanostructures to the cytoplasm of intact live cells. Here, we identify a new delivery modality, based on mechanical disruption of the plasma membrane, which efficiently mediates the delivery of nanoparticles to the cytoplasm of mammalian cells. We use two distinct execution modes, two adherent cell lines, and three sizes of semiconducting nanocrystals, or quantum dots, to demonstrate its applicability and effectiveness. As the underlying mechanism is purely physical, we anticipate that such "mechanodelivery" can be generalized to other modes of execution as well as to the cytoplasmic introduction of a structurally diverse array of functional nanomaterials.

Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts

“…The cells that brush the edge of the needle undergo membrane damage but remain adherent to the substrate. Intracellular delivery of dextrans 727 , dyes 728 , fluorescently-labeled nucleotides 729 , and quantum dots 708 has been achieved in cells adjacent to the scratch zone. Although the method is lower throughput than scrape loading, one advantage of scratch loading is that cells remain adherent for immediate analysis.…”

“…Bead loading has been used to deliver dye-conjugated dUTP for fluorescent visualization of chromosome formation, 731 antibody loading into macrophages 732,733 and fibroblasts, 734 intracellular delivery of proteins, 735−737 peptides, 738 fab fragments, 739,740 peptide nucleic acid probes, 741 SNAP-reactive dyes, 742 CNTs, 743 and quantum dots up to 15 nm in several cell lines. 744 Scrape loading has achieved intracellular delivery of proteins, 96,543,745−751 antibodies, 752−754 peptides, 755,756 morpholinos, 757 high molecular weight dextrans, 96,758 lipopolysaccharides, 759 dyes, 760,761 pH-sensitive probes, 762 and transfection of plasmids. 545 A variant of the scrape loading technique is scratch loading.…”

“…Bead and scrape loading techniques have been used to deliver a variety of cargoes into cells. Bead loading has been used to deliver dye-conjugated dUTP for fluorescent visualization of chromosome formation, antibody loading into macrophages , and fibroblasts, intracellular delivery of proteins, − peptides, fab fragments, , peptide nucleic acid probes, SNAP-reactive dyes, CNTs, and quantum dots up to 15 nm in several cell lines . Scrape loading has achieved intracellular delivery of proteins, ,,− antibodies, − peptides, , morpholinos, high molecular weight dextrans, , lipopolysaccharides, dyes, , pH-sensitive probes, and transfection of plasmids …”

A Comprehensive Review on Intracellular Delivery

“…This contact leads to the creation of sufficient strains to generate stochastic disruptions in the plasma membrane. [ 110 ] Bead loading has been used for antibody loading into fibroblasts and macrophages, [ 111 ] intracellular delivery of quantum dots (up to 15 nm), [ 112 ] proteins, and antibodies. [ 113 ] For instance, to image single mRNA translation in living cells, bead loading was used to transfer antibody fab fragments to the cell interior.…”

Label-free and live cell imaging by interferometric scattering microscopy

“… 5 , 6 On the other hand, one great advantage of fluorescence microscopy is its capability of imaging the morphological and functional alteration in live cells. 7 – 15 Nevertheless, it also has critical restrictions including limitations in observation time due to photobleaching and undesirable alteration in cellular functions induced by tagging fluorescent molecules to the specific target sites.…”