Gold Nanoparticle-Mediated Delivery of Molecules into Primary Human Gingival Fibroblasts Using ns-Laser Pulses: A Pilot Study

Judith Krawinkel,Kristian Werelius,Alexander Heisterkamp,Georgios Romanos,Maria Torres-Mapa,Susanne Gerhardt-Szép,Stefan Rüttermann

doi:10.3390/ma9050397

Abstract

Interaction of gold nanoparticles (AuNPs) in the vicinity of cells’ membrane with a pulsed laser (λ = 532 nm, τ = 1 ns) leads to perforation of the cell membrane, thereby allowing extracellular molecules to diffuse into the cell. The objective of this study was to develop an experimental setting to deliver molecules into primary human gingival fibroblasts (pHFIB-G) by using ns-laser pulses interacting with AuNPs (study group). To compare the parameters required for manipulation of pHFIB-G with those needed for cell lines, a canine pleomorphic adenoma cell line (ZMTH3) was used (control group). Non-laser-treated cells incubated with AuNPs and the delivery molecules served as negative control. Laser irradiation (up to 35 mJ/cm2) resulted in a significant proportion of manipulated fibroblasts (up to 85%, compared to non-irradiated cells: p < 0.05), while cell viability (97%) was not reduced significantly. pHFIB-G were perforated as efficiently as ZMTH3. No significant decrease of metabolic cell activity was observed up to 72 h after laser treatment. The fibroblasts took up dextrans with molecular weights up to 500 kDa. Interaction of AuNPs and a pulsed laser beam yields a spatially selective technique for manipulation of even primary cells such as pHFIB-G in high throughput.

Highlights

Molecular delivery methods are key technologies in the biomedical sciences
We describe for the first time the delivery of different molecules into primary human gingival fibroblasts using AuNPs and laser irradiation
As perforation efficiency was based on the proportion of living cells, it decreased in parallel with decreasing cell viability

Summary

Introduction

Molecular delivery methods are key technologies in the biomedical sciences. Despite several physical methodical approaches (ballistic, electricity, iontophoresis, ultrasound, light) the efficient delivery of molecules to cells remains challenging [1,2]. Lasers interacting with nanoparticles were shown to be able to efficiently deliver molecules into cells [2,4,5]. St-Louis Lalonde et al compared membrane permeabilization by irradiating AuNPs with ns-laser pulses on- (532 nm) and off- (1064 nm) resonance [5]. Another transfection technique described in literature is laser scanning of cells previously incubated with gold nanoparticles (AuNPs), called the GNOME approach. Applying the GNOME technique, Heinemann et al already described the possibility to deliver green fluorescent proteins into mammalian cells with an efficiency of 43%, while maintaining a high level of cell viability.

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