ABSTRACT
Intracellular drug delivery is at the heart of many diagnosis procedures and a key step in gene therapy. Research has been conducted to bypass cell barriers for controlled intracellular drug release and made consistent progress. However, state-of-the-art techniques based on non-viral carriers or physical methods suffer several drawbacks, including limited delivery yield, low throughput or low viability, which are key parameters in therapeutics, diagnostics and drug delivery. Nevertheless, gold nanoparticle (AuNP) mediated photoporation has stood out as a promising approach to permeabilize cell membranes through laser induced Vapour NanoBubble (VNB) generation, allowing the influx of external cargo molecules into cells. However, its use as a transfection technology for the genetic manipulation of therapeutic cells is hindered by the presence of non-degradable gold nanoparticles. Here, we report a new optofluidic method bringing gold nanoparticles in close proximity to cells for photoporation, while avoiding direct contact with cells by taking advantage of hydrodynamic focusing in a multi-flow device. Cells were successfully photoporated with [Formula: see text] efficiency with no significant reduction in cell viability at a throughput ranging from [Formula: see text] to [Formula: see text]. This optofluidic approach provides prospects of translating photoporation from an R &D setting to clinical use for producing genetically engineered therapeutic cells.
Subject(s)
Gold , Metal Nanoparticles , Humans , Pharmaceutical Preparations , Transfection , Drug Delivery SystemsABSTRACT
Surface plasmon resonance (SPR) sensing is a well-established high-sensitivity, label-free and real-time detection technique for biomolecular interaction study. Its primary working principle consists of the measurement of the optical refractive index of the medium that is in close vicinity of the sensor surface. Bio-functionalization techniques allow biomolecular events to be located in such a way. Since optical refractive indices of any medium varies with the temperature, the place where the measurement takes place shall be within a temperature-controlled environment in order to ensure any temperature fluctuation is interpreted as a biomolecular event. Since the SPR measurement probes the sensed medium within the penetration depth of the plasmonic wave, which is less or in the order of 1 µm, we propose to use the metallic film constituting the detection surface as a localized heater aiming at controlling finely and quickly the temperature of the sensed medium. The Joule heating principle is then used and the modeling of the heater is reported as well as its validation by thermal IR imaging. Using water as a demonstration medium, SPR measurement results at different temperatures are successfully compared to the theoretical optical refractive index of water versus temperature.