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Role of actin cytoskeleton in cargo delivery mediated by vertically aligned silicon nanotubes.

ABSTRACT: Nanofabrication technologies have been recently applied to the development of engineered nano-bio interfaces for manipulating complex cellular processes. In particular, vertically configurated nanostructures such as nanoneedles (NNs) have been adopted for a variety of biological applications such as mechanotransduction, biosensing, and intracellular delivery. Despite their success in delivering a diverse range of biomolecules into cells, the mechanisms for NN-mediated cargo transport remain to be elucidated. Recent studies have suggested that cytoskeletal elements are involved in generating a tight and functional cell-NN interface that can influence cargo delivery. In this study, by inhibiting actin dynamics using two drugs-cytochalasin D (Cyto D) and jasplakinolide (Jas), we demonstrate that the actin cytoskeleton plays an important role in mRNA delivery mediated by silicon nanotubes (SiNTs). Specifically, actin inhibition 12 h before SiNT-cellular interfacing (pre-interface treatment) significantly dampens mRNA delivery (with efficiencies dropping to 17.2% for Cyto D and 33.1% for Jas) into mouse fibroblast GPE86 cells, compared to that of untreated controls (86.9%). However, actin inhibition initiated 2 h after the establishment of GPE86 cell-SiNT interface (post-interface treatment), has negligible impact on mRNA transfection, maintaining > 80% efficiency for both Cyto D and Jas treatment groups. The results contribute to understanding potential mechanisms involved in NN-mediated intracellular delivery, providing insights into strategic design of cell-nano interfacing under temporal control for improved effectiveness.

SUBMITTER: Chen Y

PROVIDER: S-EPMC9461134 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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Role of actin cytoskeleton in cargo delivery mediated by vertically aligned silicon nanotubes.

Chen Yaping Y Yoh Hao Zhe HZ Shokouhi Ali-Reza AR Murayama Takahide T Suu Koukou K Morikawa Yasuhiro Y Voelcker Nicolas H NH Elnathan Roey R

Journal of nanobiotechnology 20220908 1

Nanofabrication technologies have been recently applied to the development of engineered nano-bio interfaces for manipulating complex cellular processes. In particular, vertically configurated nanostructures such as nanoneedles (NNs) have been adopted for a variety of biological applications such as mechanotransduction, biosensing, and intracellular delivery. Despite their success in delivering a diverse range of biomolecules into cells, the mechanisms for NN-mediated cargo transport remain to b ...[more]

PMID: 36076230

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Project description:Convex cylindrical silicon nanostructures, also referred to as silicon nanocones, find their value in many applications ranging from photovoltaics to nanofluidics, nanophotonics, and nanoelectronic applications. To fabricate silicon nanocones, both bottom-up and top-down methods can be used. The top-down method presented in this work relies on pre-shaping of silicon nanowires by ion beam etching followed by self-limited thermal oxidation. The combination of pre-shaping and oxidation obtains high-density, high aspect ratio, periodic, and vertically aligned sharp single-crystalline silicon nanocones at the wafer-scale. The homogeneity of the presented nanocones is unprecedented and may give rise to applications where numerical modeling and experiments are combined without assumptions about morphology of the nanocone. The silicon nanocones are organized in a square periodic lattice, with 250 nm pitch giving arrays containing 1.6 billion structures per square centimeter. The nanocone arrays were several mm2 in size and located centimeters apart across a 100-mm-diameter single-crystalline silicon (100) substrate. For single nanocones, tip radii of curvature < 3 nm were measured. The silicon nanocones were vertically aligned, baring a height variation of < 5 nm (< 1%) for seven adjacent nanocones, whereas the height inhomogeneity is < 80 nm (< 16%) across the full wafer scale. The height inhomogeneity can be explained by inhomogeneity present in the radii of the initial columnar polymer mask. The presented method might also be applicable to silicon micro- and nanowires derived through other top-down or bottom-up methods because of the combination of ion beam etching pre-shaping and thermal oxidation sharpening. A novel method is presented where argon ion beam etching and thermal oxidation sharpening are combined to tailor a high-density single-crystalline silicon nanowire array into a vertically aligned single-crystalline silicon nanocones array with < 3 nm apex radius of curvature tips, at the wafer scale.

Project description:Currently, there is a lack of ultrasensitive diagnostic tool to detect some diseases such as ischemic stroke, thereby impacting effective and efficient intervention for such diseases at an embryonic stage. In addition to the lack of proper detection of the neurological diseases, there is also a challenge in the treatment of these diseases. Carbon nanotubes have a potential to be employed in solving the theragnostic challenges in those diseases. In this study, carbon nanotubes were successfully synthesized for potential application in the detection and treatment of the neurological diseases such as ischemic stroke. Vertically aligned multiwalled carbon nanotubes (VA-MWCNTs) were purified with HCl, carboxylated with H2SO4:HNO3 (3:1) and acylated with SOCl2 for use in potential targeting studies and for the design of a carbon-based electrode for possible application in the diagnosis of neurological diseases, including ischemic stroke. MWCNTs were washed, extracted from the filter membranes and dried in a vacuum oven at 60 °C for 24 h prior to functionalization and PEGylation. CNTs were characterized by SEM, TEM, OCA, DLS, CV and EIS. The HCl-treated CNT obtained showed an internal diameter, outer diameter and thickness of 8 nm, 34 nm and 75 µm, while these parameters for the H2SO4-HNO3-treated CNT were 8 nm, 23 nm and 41µm, respectively. PEGylated CNT demonstrated zeta potential, polydispersive index and particle size distribution of 6 mV, 0.41 and 98 nm, respectively. VA-MWCNTs from quartz tube were successfully purified, carboxylated, acylated and PEGylated for potential functionalization for use in targeting studies. For designing the carbon-based electrode, VA-MWCNTs on silicon wafer were successfully incorporated into epoxy resin for diagnostic applications. Functionalized MWCNTs were nontoxic towards PC-12 neuronal cells. In conclusion, vertically super-aligned MWCNTs have been successfully synthesized and functionalized for possible theragnostic biomedical applications in neurological disorders such as ischemic stroke.

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Role of actin cytoskeleton in cargo delivery mediated by vertically aligned silicon nanotubes.

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Role of actin cytoskeleton in cargo delivery mediated by vertically aligned silicon nanotubes.

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