Elevation of plasma membrane permeability by laser irradiation of selectively bound nanoparticles

Abstract

Irradiation of nanoabsorbers with pico- and nanosecond laser pulses could result in thermal effects with a spatial confinement of less than 50 nm. Therefore absorbing nanoparticles could be used to create controlled cellular effects. We describe a combination of laser irradiation with nanoparticles, which changes the plasma membrane permeability. We demonstrate that the system enables molecules to penetrate impermeable cell membranes. Laser light at 532 nm is used to irradiate conjugates of colloidal gold, which are delivered by antibodies to the plasma membrane of the Hodgkin's disease cell line L428 and/or the human large-cell anaplastic lymphoma cell line Karpas 299. After irradiation, membrane permeability is evaluated by fluorescence microscopy and flow cytometry using propidium iodide (PI) and fluorescein isothiocyanate (FITC) dextran. The fraction of transiently permeabilized and then resealed cells is affected by the laser parameter, the gold concentration, and the membrane protein of the different cell lines to which the nanoparticles are bound. Furthermore, a dependence on particle size is found for these interactions in the different cell lines. The results suggest that after optimization, this method could be used for gene transfection and gene therapy.

Gold Nanorods Mediate Tumor Cell Death by Compromising Membrane Integrity

“…[20][21][22][23][24][25] The gradient for cavitation-induced heating declines sharply from the epicenter, such that direct thermolysis would be limited to targeted cells near the sites of photothermal transduction. [20,23,24] Cavitation is also responsible for transient bubble formation, which can expand by as much as several microns during their microsecond lifespans.…”

“…[20,23,24] Cavitation is also responsible for transient bubble formation, which can expand by as much as several microns during their microsecond lifespans. [22,33] These microbubbles have been proposed to cause a temporary rupture in cell membranes, [20,21,25] thereby increasing their permeability to EB and other chemical agents.…”

“…[17][18][19] These individual outcomes may be resolved at the subcellular level by using targeted nanoparticle delivery to administer localized photothermal effects. [20][21][22][23][24][25] For example, nanosecond laser pulses have been used to induce cavitation in cells containing gold nanoparticles, resulting in transient increases in membrane permeability and inactivation of adsorbed proteins. [20,21] These processes are quite distinct from those based on systemic changes in temperature.…”

Gold nanoparticle-mediated laser stimulation causes a complex stress signal in neuronal cells

“…• Several studies have suggested that while the rate of ion inflow is highest during the first ~10s after laser irradiation, pores can remain open for minutes 1,13 . Assuming that the large Ca 2+ gradient across the membrane cannot be balanced immediately (due to the energy barrier within the pore), a proportion of cells should be expected to show fast influx during the first seconds that would then be slowed down when the first pores close, and continue to approximate the extracellular calcium concentration.…”

Mechanisms of laser nanoparticle-based techniques for gene transfection—a calculation study

“…In our laboratory, Yao and co-workers have demonstrated through experimental results that the plasma membrane can be transiently permeabilized with high efficiency when 30-nm gold nanoparticles are bound to membrane proteins and irradiated with nanosecond pulses [3]. Based on this technique, they have successfully transferred small exogenous molecules into cells.…”