Nanoparticle-sensitized photoporation is an upcoming approach for intracellular delivery of biologics, combining high efficiency and throughput with excellent cell viability. However, as it relies on close contact between nanoparticles and cells, its translation towards clinical applications is hampered by safety and regulatory concerns. Here, we show that light-sensitive iron oxide nanoparticles (IONPs) embedded in biocompatible electrospun nanofibers induce membrane permeabilization by photothermal effects without direct cellular contact with IONPs. The photothermal nanofibers are successfully used to deliver effector molecules, including CRISPR/Cas9 ribonucleoprotein complexes and siRNA, in adherent and suspension cells, including embryonic stem cells and hard-to-transfect T-cells without affecting cell proliferation or phenotype.
In vivo
experiments furthermore demonstrate successful tumor regression in mice treated with CAR-T cells in which expression of PD1 is downregulated after nanofiber photoporation with siPD1. In conclusion, cell membrane permeabilization with photothermal nanofibers is a promising concept towards the safe and more efficient production of engineered cells for therapeutic applications, including stem cell or adoptive T cell therapy.
The
analytical figures of merit of a low-dispersion (ultrafast)
ablation cell geometry within the Cobalt ablation chamber, integrated
into a nanosecond laser ablation–inductively coupled plasma-mass
spectrometer system, are reported. The system was investigated for
its capability for fast high-resolution elemental imaging. A spot
of 0.6 μm diameter was achieved on the sample surface by aperture
imaging of a 10 μm pinhole. The resulting conical crater (0.6
μm ⌀ × 130 nm↓) morphology in a Au-coated
glass target and carbon-coated silica wafer was characterized with
atomic force microscopy. The Cobalt ablation chamber is based around
a motorized height-adjustable tube cell, which allows modulating the
sampling distance, i.e. the distance between the sample surface and
the cell inlet, in a dynamic manner. This distance was observed to
influence the single pulse response profile. The variation of the
average signal intensity at multiple sample heights within a range
of 0.5 mm was <3% RSD. Under optimum conditions, single pulse responses
with a full width at 10% of the maximum peak intensity (FW0.1M) of
∼1 ms can be achieved for 238U upon ablation of
NIST SRM612 glass, effectively opening the way to pixel acquisition
rates up to 1 kHz. To demonstrate the potential of this technology,
the elemental distribution of Zn in small intestine villi of mice
subjected to a Zn-enriched diet was imaged using the 0.6 μm
spot size, and rapid imaging of a zircon grain cross-section was performed.
This work reports on the analytical figures of merit of a low-dispersion aerosol transport system for high-throughput bulk and spatially resolved analysis via LA-ICP-MS. This device maximizes the collection of...
Biomimetic functionalization to confer stealth and targeting properties to nanoparticles is a field of intense study. Extracellular vesicles (EV), sub-micron delivery vehicles for intercellular communication, have unique characteristics for drug delivery. We investigated the top-down functionalization of gold nanoparticles with extracellular vesicle membranes, including both lipids and associated membrane proteins, through mechanical extrusion. EV surface-exposed membrane proteins were confirmed to help avoid unwanted elimination by macrophages, while improving autologous uptake. EV membrane morphology, protein composition and orientation were found to be unaffected by mechanical extrusion. We implemented complementary EV characterization methods, including transmission- and immune-electron microscopy, and nanoparticle tracking analysis, to verify membrane coating, size and zeta potential of the EV membrane-cloaked nanoparticles. While successful EV membrane coating of the gold nanoparticles resulted in lower macrophage uptake, low yield was found to be a significant downside of the extrusion approach. Our data incentivize more research to leverage EV membrane biomimicking as a unique drug delivery approach in the near future.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.