| Autor: |
Illath K; Department of Engineering Design, Indian Institute of Technology Madras, India. santra.tuhin@gmail.com., Kar S; Department of Physics, Indian Institute of Science Education and Research, Tirupati, India., Shinde A; Department of Engineering Design, Indian Institute of Technology Madras, India. santra.tuhin@gmail.com., Ojha R; Department of Physical Medicine and Rehabilitation, Christian Medical College, Vellore, India., Iyer DR; Department of Biotechnology, Indian Institute of Technology Madras, India., Mahapatra NR; Department of Biotechnology, Indian Institute of Technology Madras, India., Nagai M; Department of Mechanical Engineering, Toyohashi University of Technology, Aichi, Japan., Santra TS; Department of Engineering Design, Indian Institute of Technology Madras, India. santra.tuhin@gmail.com. |
| Abstrakt: |
Photothermal nanoparticle-sensitised photoporation is an emerging approach, which is considered an efficient tool for the intracellular delivery of biomolecules. Nevertheless, using this method to achieve high transfection efficiency generally compromises cell viability and uneven distribution of nanoparticles results in non-uniform delivery. Here, we show that high aspect ratio gold nano-burflowers, synthesised in a microfluidic device, facilitate highly efficient small to very-large cargo delivery uniformly using infrared light pulses without sacrificing cell viability. By precisely controlling the flow rates of shaping reagent and reducing agent, high-density (24 numbers) sharply branched spikes (∼80 nm tip-to-tip length) of higher aspect ratios (∼6.5) with a small core diameter (∼45 nm) were synthesised. As produced gold burflower-shape nanoparticles are biocompatible, colloidally stable (large surface zeta potential value), and uniform in morphology with a higher plasmonic peak (max. 890 nm). Theoretical analysis revealed that spikes on the nanoparticles generate a higher electromagnetic field enhancement upon interaction with light pulses. It induces plasmonic nanobubbles in the vicinity of the cells, followed by pore formation on the membrane leading to diverse biomolecular delivery into cells. Our platform has been successfully implemented for uniform delivery of small to very large biomolecules, including siRNA (20-24 bp), plasmid DNA expressing green fluorescent protein (6.2 kbp), Cas-9 plasmid (9.3 kbp), and β-galactosidase enzyme (465 kDa) into diverse mammalian cells with high transfection efficiency and cell viability. For very large biomolecules such as enzymes, the best results were achieved as ∼100% transfection efficiency and ∼100% cell viability in SiHa cells. Together, our findings demonstrate that the spiky gold nano-burflower shape nanoparticles manufactured in a microfluidic system exhibited excellent plasmonic behaviour and could serve as an effective tool in manipulating cell physiology. |
