Biophysical and Biochemical Regulation of Cell Dynamics in Magnetically Assembled Cellular Structures.

Soluble signaling molecules and extracellular matrix (ECM) regulate cell dynamics in various biological processes. Wound healing assays are widely used to study cell dynamics in response to physiological stimuli. However, traditional scratch-based assays can damage the underlying ECM-coated substrates. Here, we use a rapid, non-destructive, label-free magnetic exclusion technique to form annular aggregates of bronchial epithelial cells on tissue-culture treated (TCT) and ECM-coated surfaces within 3 h. The cell-free areas enclosed by the annular aggregates are measured at different times to assess cell dynamics. The effects of various signaling molecules, including epidermal growth factor (EGF), oncostatin M, and interleukin 6, on cell-free area closures are investigated for each surface condition. Surface characterization techniques are used to measure the topography and wettability of the surfaces. Further, we demonstrate the formation of annular aggregates on human lung fibroblast-laden collagen hydrogel surfaces, which mimic the native tissue architecture. The cell-free area closures on hydrogels indicate that the substrate properties modulate EGF-mediated cell dynamics. The magnetic exclusion-based assay is a rapid and versatile alternative to traditional wound healing assays.

SARS-CoV-2 detection with aptamer-functionalized gold nanoparticles.

A rapid detection test for SARS-CoV-2 is urgently required to monitor virus spread and containment. Here, we describe a test that uses nanoprobes, which are gold nanoparticles functionalized with an aptamer specific to the spike membrane protein of SARS-CoV-2. An enzyme-linked immunosorbent assay confirms aptamer binding with the spike protein on gold surfaces. Protein recognition occurs by adding a coagulant, where nanoprobes with no bound protein agglomerate while those with sufficient bound protein do not. Using plasmon absorbance spectra, the nanoprobes detect 16 nM and higher concentrations of spike protein in phosphate-buffered saline. The time-varying light absorbance is examined at 540 nm to determine the critical coagulant concentration required to agglomerates the nanoprobes, which depends on the protein concentration. This approach detects 3540 genome copies/µl of inactivated SARS-CoV-2.

Fibroblasts Accelerate Formation and Improve Reproducibility of 3D Cellular Structures Printed with Magnetic Assistance.

Fibroblasts (mouse, NIH/3T3) are combined with MDA-MB-231 cells to accelerate the formation and improve the reproducibility of 3D cellular structures printed with magnetic assistance. Fibroblasts and MDA-MB-231 cells are cocultured to produce 12.5 : 87.5, 25 : 75, and 50 : 50 total population mixtures. These mixtures are suspended in a cell medium containing a paramagnetic salt, Gd-DTPA, which increases the magnetic susceptibility of the medium with respect to the cells. A 3D monotypic MDA-MB-231 cellular structure is printed within 24 hours with magnetic assistance, whereas it takes 48 hours to form a similar structure through gravitational settling alone. The maximum projected areas and circularities, and cellular ATP levels of the printed structures are measured for 336 hours. Increasing the relative amounts of the fibroblasts mixed with the MDA-MB-231 cells decreases the time taken to form the structures and improves their reproducibility. Structures produced through gravitational settling have larger maximum projected areas and cellular ATP, but are deemed less reproducible. The distribution of individual cell lines in the cocultured 3D cellular structures shows that printing with magnetic assistance yields 3D cellular structures that resemble in vivo tumors more closely than those formed through gravitational settling. The results validate our hypothesis that (1) fibroblasts act as a "glue" that supports the formation of 3D cellular structures, and (2) the structures are produced more rapidly and with greater reproducibility with magnetically assisted printing than through gravitational settling alone. Printing of 3D cellular structures with magnetic assistance has applications relevant to drug discovery, lab-on-chip devices, and tissue engineering.

Label-Free Magnetic-Field-Assisted Assembly of Layer-on-Layer Cellular Structures.

Controlled cell assembly is essential for fabricating in vitro 3D models that mimic the physiology of in vivo cellular architectures. Whereas tissue engineering techniques often rely on intrusive magnetic nanoparticles placed in cells and hydrogel encapsulation of cells to produce multilayered cellular constructs, we describe a high-throughput, label-free, and scaffold-free magnetic field-guided technique that assembles cells into a layered aggregate. An inhomogeneous magnetic field influences the diamagnetic cells suspended in a paramagnetic culture medium. Driven by the magnetic susceptibility difference and the field gradient, the cells are displaced toward the region of lowest field strength. Two cell lines are used to demonstrate the sequential assembly of layer-on-layer aggregates in microwells within 6 h. The effect of magnet size on the assembly dynamics is characterized and a microwell size criterion for the highest cell aggregation provided. Label-free magnetic-field-assisted assembly is relevant for on-demand scalable biofabrication of complex layered structures. Potential applications include drug discovery, developmental biology, lab-on-chip devices, and cancer research.

Gadopentatic acid affects in vitro proliferation and doxorubicin response in human breast adenocarcinoma cells.

Contrasting agents (CAs) that are administered to patients during magnetic resonance imaging to facilitate tumor identification are generally considered harmless. However, gadolinium (Gd) based contrast agents can be retained in the body, inflicting specific cell line cytotoxicity. We investigate the effect of Gadopentatic acid (Gd-DTPA) on human breast adenocarcinoma MCF-7 cells. These cells exhibit a toggle switch response: exposure to 0.1 and 1 mM concentrations of Gd-DTPA enhances proliferation, which is hindered at a higher 10 mM concentration. Proliferation is enhanced when cells transition to 3D morphologies in post confluent conditions. The proliferation dependence on the concentration of CA is absent for Hs 578T and MDA-MB-231 triple negative cell lines. MCF-7 cells reveal a double toggle switch related to the expression of VEGF, which goes through high-low-high downregulation when cells are exposed to 0.1, 1, and 10 mM Gd-DTPA, respectively. Finally, doxorubicin drug response is assessed, which also reveals a double toggle switch behavior, where drug cytotoxicity exhibits a nonlinear dependence on the CA concentration. A toggle switch in cell characteristics that are exposed to 1 mM of Gd-DTPA amplifies the importance of this threshold, affecting several cell behaviors if surpassed. This work emphasizes the important effects that CAs can have on cells, specifically Gd-DTPA on MCF-7 cells, and the implications for cell growth and drug response during clinical and synthetic biology procedures.

3D cellular structures and co-cultures formed through the contactless magnetic manipulation of cells on adherent surfaces.

A magnet array is employed to manipulate diamagnetic cells that are contained in paramagnetic medium to demonstrate for the first time the contactless bioprinting of three-dimensional (3D) cellular structures and co-cultures of breast cancer MCF-7 and endothelial HUVEC at prescribed locations on tissue culture treated well plates. Sequential seeding of different cell lines and the spatial displacement of the magnet array creates co-cultured cellular structures within a well without using physically intrusive well inserts. Both monotypic and co-culture experiments produce morphologically rich 3D cell structures that are otherwise absent in regular monolayer cell cultures. The magnetic contactless bioprinting of cells provides further insight into cell behaviour, invasion strategies and transformations that are useful for potential applications in drug screening, 3D cell culture formation and tissue engineering.

Swing-like pool boiling on nano-textured surfaces for microgravity applications related to cooling of high-power microelectronics.

Here, we demonstrate that heat removed in pool boiling from a heater mimicking high-power microelectronics could be used to facilitate a swing-like motion of the heater before being finally dissipated. This swing-like motion could be beneficial for shedding a large vapor bubble that encapsulates high-power heaters in microgravity where buoyancy force is unavailable for vapor bubble removal. The swing-like motion is propelled by vapor bubble recoil, the force which exists irrespective of gravity and buoyancy. We also demonstrate that this force could be significantly enhanced by depositing on the heater surface supersonically blown polymer nanofibers with cross-sectional diameters below 100 nm. These nanofibers provide additional nucleation sites, resulting in much more frequent bubble nucleation and departure, and thus a higher overall vapor recoil force propelling the heater motion. Such nanofibers strongly adhere to the heater surface and withstand prolonged harsh pool boiling. The measured velocity of the model swing-like heater in Novec 7300 fluid is about 1 cm/s.

Nickel oxide nanotube synthesis using multiwalled carbon nanotubes as sacrificial templates for supercapacitor application.

A novel approach for the fabrication of nickel oxide nanotubes based on multiwalled carbon nanotubes as a sacrificial template is described. Electroless deposition is employed to deposit nickel onto carbon nanotubes. The subsequent annealing of the product in the presence of air oxidizes nickel to nickel oxide, and carbon is released as gaseous carbon dioxide, leaving behind nickel oxide nanotubes. Electron microscopy and elemental mapping confirm the formation of nickel oxide nanotubes. New chelating polyelectrolytes are used as dispersing agents to achieve high colloidal stability for both the nickel-coated carbon nanotubes and the nickel oxide nanotubes. A gravimetric specific capacitance of 245.3 F g-1 and  an areal capacitance of 3.28 F cm-2 at a scan rate of 2 mV s-1 is achieved, with an electrode fabricated using nickel oxide nanotubes as the active element with a mass loading of 24.1 mg cm-2.

Dynamic Study of Liquid Drop Impact on Supercooled Cerium Dioxide: Anti-Icing Behavior.

This work deals with the anti-icing behavior at subfreezing temperatures of CeO2/polyurethane nanocomposite coatings with and without a stearic acid treatment on aluminum alloy substrates. The samples ranged from superhydrophilic to superhydrophobic depending on surface morphology and surface functionalization. X-ray photoelectron spectroscopy was used to determine the surface composition. The anti-icing behavior was studied both by importing fog into a chamber with controlled atmosphere at subzero temperatures and by conducting experiments with drop impact velocities of 1.98, 2.8, 3.83, and 4.95 m/s. It was found that the ice-phobicity of the ceramic/polymer nanocomposite coating was dependent on the surface roughness and surface energy. Water drops were observed to completely rebound from the surface at subfreezing temperatures from superhydrophobic surfaces with small contact angle hysteresis regardless of the impact velocity, thus revealing the anti-icing capability of such surfaces.

In Situ 3D Label-Free Contactless Bioprinting of Cells through Diamagnetophoresis.

Using whole blood, we demonstrate the first realization of a novel macroscale, contactless, label-free method to print in situ three-dimensional (3D) cell assemblies of different morphologies and sizes. This novel bioprinting method does not use nozzles that can contaminate the cell suspension, or to which cells can adhere. Instead, we utilize the intrinsic diamagnetic properties of whole blood cells to magnetically manipulate them in situ in a nontoxic paramagnetic medium, creating (a) rectangular bar, (b) three-pointed star, and (c) spheroids of varying sizes. We envision the technique to be transferable to other cell lines, with potential applications in tissue engineering and drug screening.