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Triplett, Michael

Publications and source records attributed to Triplett, Michael.

Rapid curing dynamics of PEG-thiol-ene resins allow facile 3D bioprinting and in-air cell-laden microgel fabrication

Thiol-norbornene photoclick hydrogels are highly efficient in tissue engineering applications due to their fast gelation, cytocompatibility, and tunability. In this work, we utilized the advantageous features of polyethylene glycol (PEG)-thiol-ene resins to enable fabrication of complex and heterogeneous tissue scaffolds using 3D bioprinting and in-air drop encapsulation techniques. We demonstrated that photoclickable PEG-thiol-ene resins could be tuned by varying the ratio of PEG-dithiol to PEG norbornene to generate a wide range of mechanical stiffness (0.5–12 kPa) and swelling ratios. Importantly, all formulations maintained a constant, rapid gelation time (<0.5 s). We used this resin in biological projection microstereolithography (BioPµSL) to print complex structures with geometric fidelity and demonstrated biocompatibility by printing cell-laden microgrids. Moreover, the rapid gelling kinetics of this resin permitted high-throughput fabrication of tunable, cell-laden microgels in air using a biological in-air drop encapsulation apparatus (BioIDEA). We demonstrated that these microgels could support cell viability and be assembled into a gradient structure. This PEG-thiol-ene resin, along with BioPµSL and BioIDEA technology, will allow rapid fabrication of complex and heterogeneous tissues that mimic native tissues with cellular and mechanical gradients. The engineered tissue scaffolds with a controlled microscale porosity could be utilized in applications including gradient tissue engineering, biosensing, and in vitro tissue models.

36 MATERIALS SCIENCE↗

Waveguides for neurostimulation in the cochlea

Introduction Optical stimulation has been suggested for neural stimulation to improve cochlear implants. Light allows for more spatially selective activation of neuron populations than electrical current, offering more independent frequency bands along the spiral ganglion. These bands are available to encode acoustic information with anticipated better frequency resolution, improving cochlear implant user performance in noisy listening environments, tonal languages, and music perception. Methods Optical cochlear implants (oCIs) can deliver light either directly via small emitters within the cochlea or via waveguides from external optical sources. We investigated three waveguide designs made from OrmoComp ® , a polymer that cures through ultraviolet (UV) radiation. Waveguides were fabricated via injection molding and coated using dip-coating or thermal reflow, or through aspiration of OrmoComp ® into polyimide tubing that served as the cladding of the waveguide. The choice of fabrication technique directly determined the waveguides' total diameter: thermal reflow yielded ≈940 μm, dip-coating produced ≈306 μm, and aspiration resulted in ≈132 μm core diameter waveguides. Given the human cochlea's small size, we focused on analyzing the 306-μm and 132-μm waveguides, evaluating their optical performance (propagation and bending losses) and mechanical properties (bending stiffness and insertion forces). Furthermore, we evaluated some of these designs in in-vivo guinea pigs experiments. Results For the 100-μm core diameter waveguides, the propagation losses were 12.34 ± 1.26, 1.18 ± 0.88, 1.49 ± 0.58, and 3.43 ± 0.68 dB/cm at 534, 1,375, 1,460, and 1,550 nm, respectively. The respective bending losses at a 2 mm radius of curvature were 5.50 ± 1.32, 0.56 ± 0.26, 0.79 ± 0.18, and 0.64 ± 0.23 dB, and at 1 mm 8.54 ± 1.30, 2.05 ± 0.84, 2.11 ± 0.50, and 1.44 ± 0.37 dB. The bending stiffness of a 1 mm segment of the 100-μm-diameter waveguides was 18.9 ± 2.2 N/m. Insertion forces for the 100-μm-diameter waveguides into an acrylic human-size scala tympani model were < 25 mN. For the waveguides with 306 and 940 μm total diameter, the propagation losses ranged between 0.43 and 2.40 dB/cm at 534, 680, 1,375, and 1,550 nm, between 2.19 and 3.78 dB/cm at 450 and 1,460 nm. Bending losses for 360 degrees at 1,375 nm were 5.0, 2.4, and 0.46 for a bending radius of 2.5-, 3-, and 4-mm. Discussion Our study demonstrated that the polymer OrmoComp ® is suitable for fabricating waveguides to transmit near-infrared radiation. In-vivo experiments showed optically evoked auditory responses originating from optical stimulation in the guinea pigs' first cochlear turn while radiation was delivered. Incoming experiments will focus on evaluating long-term performance of these waveguides in guinea pigs and cats. This study will be designed to provide insights into the waveguides' performance and biocompatibility over extended periods, essential for their potential clinical application in future oCIs.

Kim, Matthew↗

Spatiotemporal analysis of 3D human iPSC-derived neural networks using a 3D multi-electrode array

While there is a growing appreciation of three-dimensional (3D) neural tissues (i.e., hydrogel-based, organoids, and spheroids), shown to improve cellular health and network activity to mirror brain-like activity in vivo , functional assessment using current electrophysiology techniques (e.g., planar multi-electrode arrays or patch clamp) has been technically challenging and limited to surface measurements at the bottom or top of the 3D tissue. As next-generation MEAs, specifically 3D MEAs, are being developed to increase the spatial precision across all three dimensions (X, Y, Z), development of improved computational analytical tools to discern region-specific changes within the Z dimension of the 3D tissue is needed. In the present study, we introduce a novel computational analytical pipeline to analyze 3D neural network activity recorded from a “bottom-up” 3D MEA integrated with a 3D hydrogel-based tissue containing human iPSC-derived neurons and primary astrocytes. Over a period of ~6.5 weeks, we describe the development and maturation of 3D neural activity (i.e., features of spiking and bursting activity) within cross sections of the 3D tissue, based on the vertical position of the electrode on the 3D MEA probe, in addition to network activity (identified using synchrony analysis) within and between cross sections. Then, using the sequential addition of postsynaptic receptor antagonists, bicuculline (BIC), 2-amino-5-phosphonovaleric acid (AP-5), and 6-cyano-5-nitroquinoxaline-2,3-dione (CNQX), we demonstrate that networks within and between cross sections of the 3D hydrogel-based tissue show a preference for GABA and/or glutamate synaptic transmission, suggesting differences in the network composition throughout the neural tissue. The ability to monitor the functional dynamics of the entire 3D reconstructed neural tissue is a critical bottleneck; here we demonstrate a computational pipeline that can be implemented in studies to better interpret network activity within an engineered 3D neural tissue and have a better understanding of the modeled organ tissue.

3D culture↗

System and method for three-dimensional in vitro flexible microelectrode array

The present disclosure relates to a biocompatible, in vitro probe system. The probe system may have a substrate and a culture well supported on the substrate. The culture well defines a three-dimensional volume for containing in vitro cultures of electroactive cells. The probe system has at least one probe subsystem supported on the substrate. The probe subsystem has at least one probe having an array of electrodes, with the probe being disposed within the culture well for in vitro electrically communicating with the electroactive cells. The probe subsystem is adapted to be interfaced to an external instrumentation/recording device.

Soscia, David↗