A novel cell transfection platform based on laser optoporation mediated by Au nanostar layers

Abstract

The recently developed laser-induced cell transfection mediated by Au nanoparticles is a promising alternative to the well-established lipid-based transfection or to electroporation. Optoporation is based on the laser plasmonic heating of nanoparticles located near the cell membrane. However, the uncontrollable cell damage from intense laser pulses and from random attachment of nanoparticles may be crucial for transfection. We present a novel plasmonic optoporation technique that uses Au nanostar layers immobilized in culture microplate wells. HeLa cells were grown directly on Au nanostar layers, after which they were subjected to continuous-wave 808 nm laser irradiation. An Au monolayer density ~15 μg/cm and an absorbed energy of about 15 to 30 J were found to be optimal for optoporation. Propidium iodide molecules were used as model penetrating agent. The transfection efficiency evaluated using fluorescence microscopy for HeLa cells transfected with pGFP under optimized optoporation conditions (95% ± 5%) was similar to the efficiency of TurboFect. The technique's efficiency (295 ± 10 relative light units, RLU), demonstrated by transfecting HeLa cells with the pCMV-GLuc 2 control plasmid, was greater than that obtained by transfection of HeLa cells with the TurboFect agent (220 ± 10 RLU). The cell viability in plasmonic optoporation (92% ± 7%), too, was greater than that in transfection with TurboFect (75% ± 7%).

Graphene quantum dot based charge‐reversal nanomaterial for nucleus‐targeted drug delivery and efficiency controllable photodynamic therapy

“…As small pieces of graphene, graphene quantum dots (GQDs) have received significant attention in biosensing and bioimaging, owing to their tunable photoluminescence (PL) and good biocompatibility as well as photostability . Interestingly, GQD has been successfully used as a drug delivery system for the pH‐sensitive release of anti‐cancer drugs , attributing to the fact that such small‐size (<50 nm) drug carriers are more likely to accumulate in tumors .…”

Polydopamine-coated Au nanorods for targeted fluorescent cell imaging and photothermal therapy

“…A temperature of 60 °C in the well is sufficient to obtain complete cell death after a dozen of seconds [48]. On the other hand, the efficiency of photothermal treatment is usually connected to a local increase in temperature around cancer cells or tissues without heating of the solution or surrounding tissues.…”

Engineered Gold-Based Nanomaterials: Morphologies and Functionalities in Biomedical Applications. A Mini Review

“…As is well known, morphology plays a key role in gold-nanotechnology, especially in relation to the possibility of having materials with different symmetries (spheres, rods, and cubes) [1,2,3,4,12,24,28,70,71,72,73,74] or without a defined and reproducible symmetry (stars, hollow particles, and cages) [1,75,76,77,78,79,80,81,82,83,84]. Highly symmetrical particles are the most studied because of their low dispersity and synthetic reproducibility, which assure the repeatability of experiments and results.…”

“…This review shows the advantages and disadvantages of the different morphologies, considering the dimension, functionalization and bioapplication. In this regard, in the wide biomedicine field, four main areas of applications are typically identified: therapy, diagnostics, drug/gene delivery, and sensors, as schematically shown in Figure 2 [4,8,10,12,16,17,18,21,28,29,30,31,32,33,34,37,38,39,48,51,53,70,71,72,75,76,77,78,79,80,81,82,83,84,85,90,93,94,95,96,97,98,99,100,101,102,103,104,105,106,107,108,109,110,111,112,113,…”