title: Comparison of gold nanoparticle mediated photoporation: vapor nanobubbles outperform direct heating for delivering macromolecules in live cells.
abstract: There is a great interest in delivering macromolecular agents into living cells for therapeutic purposes, such as siRNA for gene silencing. Although substantial effort has gone into designing nonviral nanocarriers for delivering macromolecules into cells, translocation of the therapeutic molecules from the endosomes after endocytosis into the cytoplasm remains a major bottleneck. Laser-induced photoporation, especially in combination with gold nanoparticles, is an alternative physical method that is receiving increasing attention for delivering macromolecules in cells. By allowing gold nanoparticles to bind to the cell membrane, nanosized membrane pores can be created upon pulsed laser illumination. Depending on the laser energy, pores are created through either direct heating of the AuNPs or by vapor nanobubbles (VNBs) that can emerge around the AuNPs. Macromolecules in the surrounding cell medium can then diffuse through the pores directly into the cytoplasm. Here we present a systematic evaluation of both photoporation mechanisms in terms of cytotoxicity, cell loading, and siRNA transfection efficiency. We find that the delivery of macromolecules under conditions of VNBs is much more efficient than direct photothermal disturbance of the plasma membrane without any noticeable cytotoxic effect. Interestingly, by tuning the laser energy, the pore size could be changed, allowing control of the amount and size of molecules that are delivered in the cytoplasm. As only a single nanosecond laser pulse is required, we conclude that VNBs are an interesting photoporation mechanism that may prove very useful for efficient high-throughput macromolecular delivery in live cells.
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Ghent University , Harelbekestraat 72, 9000 Ghent, Belgium.
authors.forename: Ranhua
authors.initials: R
authors.lastname: Xiong
authors.forename: Koen
authors.initials: K
authors.lastname: Raemdonck
authors.forename: Karen
authors.initials: K
authors.lastname: Peynshaert
authors.forename: Ine
authors.initials: I
authors.lastname: Lentacker
authors.forename: Ine
authors.initials: I
authors.lastname: De Cock
authors.forename: Jo
authors.initials: J
authors.lastname: Demeester
authors.forename: Stefaan C
authors.initials: SC
authors.lastname: De Smedt
authors.forename: Andre G
authors.initials: AG
authors.lastname: Skirtach
authors.forename: Kevin
authors.initials: K
authors.lastname: Braeckmans
mesh_terms: D000327:Adsorption; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival; D003593:Cytoplasm / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D006046:Gold / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006358:Hot Temperature; D006801:Humans; D007834:Lasers; D046911:Macromolecular Substances; D008567:Membranes, Artificial; D053768:Metal Nanoparticles / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry; D036103:Nanotechnology; D010777:Photochemistry; D034741:RNA, Small Interfering / Q000378:metabolism
chemical_list: D046911:Macromolecular Substances; D008567:Membranes, Artificial; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; D006046:Gold