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Research Experiences for Undergraduates (REU) sites widely serve as the first major research gateway for undergraduates seeking a structured research experience. Given their lack of prior research skills, and the highly compressed duration of the REU programs, these students frequently encounter barriers to a seamless transition into a new laboratory environment. We hypothesized that the design of a unified short course on laboratory and analysis techniques could serve as a pivotal orientation experience. Our goal was to rapidly align student expertise to their summer research goals while also integrating the student participants into a cohesive learning community. This article discusses the design and outcomes of a Cellular Bioengineering Boot Camp, which is offered at the outset of the 10-week REU site at Rutgers. The Boot Camp provides hands-on, supervised training for techniques and procedures that are common among projects. The training establishes a common language and baseline for the REU students and allows their first laboratory experiences to be with each other, and creates an immediate network of peers and mentors. Surveys before and after the Boot Camp and at the end of the summer indicated a significant improvement in student proficiency in the techniques that was retained throughout the summer. We believe that the Boot Camp approach can be tailored to the specifics of each REU site and its associated projects and research foci.

SARS-CoV-2 is the third pathogenic coronavirus to emerge since 2000. Experience from prior outbreaks of SARS-CoV and MERS-CoV has demonstrated the importance of both humoral and cellular immunity to clinical outcome, precepts that have been recapitulated for SARS-CoV-2. Despite the unprecedented rapid development and deployment of vaccines against SARS-CoV-2, more vaccines are needed to meet global demand and to guard against immune evasion by newly emerging SARS-CoV-2 variants. Here we describe the development of pGO-1002, a novel bi-cistronic synthetic DNA vaccine that encodes consensus sequences of two SARS-CoV-2 antigens, Spike and ORF3a. Mice immunized with pGO-1002 developed humoral and cellular responses to both antigens, including antibodies and capable of neutralizing infection by a clinical SARSCoV-2 isolate. Rats immunized with pGO-1002 by intradermal (ID) injection followed by application of suction with our GeneDerm device also developed humoral responses that included neutralizing antibodies and RBD-ACE2 blocking antibodies as well as robust cellular responses to both antigens. Significantly, in a Syrian hamster vaccination and challenge model, ID+GeneDerm-assisted vaccination prevented viral replication in the lungs and significantly reduced viral replication in the nares of hamsters challenged with either an ancestral SARS-CoV-2 strain or the B.1.351 (Beta) variant of concern. Furthermore, vaccinated immune sera inhibited virus-mediated cytopathic effects in vitro. These data establish the immunogenicity of the SARS-CoV-2 vaccine candidate pGO-1002 which induces potent humoral and cellular responses to the Spike and ORF3a antigens and may provide greater protection against emerging variants. [ABSTRACT FROM AUTHOR]

Abstract Background During traditional acupuncture therapy, soft tissues attach to and wind around the acupuncture needle. To study this phenomenon in a controlled and quantitative setting, we performed acupuncture needling in vitro. Methods Acupuncture was simulated in vitro in three-dimensional, type I collagen gels prepared at 1.5 mg/ml, 2.0 mg/ml, and 2.5 mg/ml collagen, and either crosslinked with formalin or left untreated. Acupuncture needles were inserted into the gels and rotated via a computer-controlled motor at 0.3 rev/sec for up to 10 revolutions while capturing the evolution of birefringence under cross-polarization. Results Simulated acupuncture produced circumferential alignment of collagen fibers close to the needle that evolved into radial alignment as the distance from the needle increased, which generally matched observations from published tissue explant studies. All gels failed prior to 10 revolutions, and the location of failure was near the transition between circumferential and radial alignment. Crosslinked collagen failed at a significantly lower number of revolutions than untreated collagen, whereas collagen concentration had no effect on gel failure. The strength of the alignment field increased with increasing collagen concentration and decreased with crosslinking. Separate studies were performed in which the gel thickness and depth of needle insertion were varied. As gel thickness increased, gels failed at fewer needle revolutions. For the same depth of insertion, alignment was greater in thinner gels. Alignment increased as the depth of insertion increased. Conclusion These results indicate that the mechanostructural properties of soft connective tissues may affect their response to acupuncture therapy. The in vitro model provides a platform to study mechanotransduction during acupuncture in a highly controlled and quantitative setting.

Single-unit recording neural probes have significant advantages towards improving signal-to-noise ratio and specificity for signal acquisition in brain-to-computer interface devices. Long-term effectiveness is unfortunately limited by the chronic injury response, which has been linked to the mechanical mismatch between rigid probes and compliant brain tissue. Small, flexible microelectrodes may overcome this limitation, but insertion of these probes without buckling requires supporting elements such as a stiff coating with a biodegradable polymer. For these coated probes, there is a design trade-off between the potential for successful insertion into brain tissue and the degree of trauma generated by the insertion. The objective of this study was to develop and validate a finite element model (FEM) to simulate insertion of coated neural probes of varying dimensions and material properties into brain tissue. Simulations were performed to predict the buckling and insertion forces during insertion of coated probes into a tissue phantom with material properties of brain. The simulations were validated with parallel experimental studies where probes were inserted into agarose tissue phantom, ex vivo chick embryonic brain tissue, and ex vivo rat brain tissue. Experiments were performed with uncoated copper wire and both uncoated and coated SU-8 photoresist and Parylene C probes. Model predictions were found to strongly agree with experimental results (<10% error). The ratio of the predicted buckling force-to-predicted insertion force, where a value greater than one would ideally be expected to result in successful insertion, was plotted against the actual success rate from experiments. A sigmoidal relationship was observed, with a ratio of 1.35 corresponding to equal probability of insertion and failure, and a ratio of 3.5 corresponding to a 100% success rate. This ratio was dubbed the "safety factor", as it indicated the degree to which the coating should be over-designed to ensure successful insertion. Probability color maps were generated to visually compare the influence of design parameters. Statistical metrics derived from the color maps and multi-variable regression analysis confirmed that coating thickness and probe length were the most important features in influencing insertion potential. The model also revealed the effects of manufacturing flaws on insertion potential. [ABSTRACT FROM AUTHOR]

Mechanical damage to axons is a proximal cause of deficits following traumatic brain injury and spinal cord injury. Axons are injured predominantly by tensile strain, and identifying the strain experienced by axons is a critical step toward injury prevention. White matter demonstrates complex nonlinear mechanical behavior at the continuum level that evolves from even more complex, dynamic, and composite behavior between axons and the “glial matrix” at the microlevel. In situ, axons maintain an undulated state that depends on the location of the white matter and the stage of neurodevelopment. When exposed to tissue strain, axons do not demonstrate pure affine or non-affine behavior, but instead transition from non-affine-dominated kinematics at low stretch levels to affine kinematics at high stretch levels. This transitional and predominant kinematic behavior has been linked to the natural coupling of axons to each other via the glial matrix. In this paper, a transitional kinematic model is applied to a micromechanics finite element model to simulate the axonal behavior within a white matter tissue subjected to uniaxial tensile stretch. The effects of the transition parameters and the volume fraction of axons on axonal behavior is evaluated and compared to previous experimental data and numerical simulations. [ABSTRACT FROM AUTHOR]

Extracellular matrix provides critical signaling context to resident cells through mechanical and bioactive properties. To realize the potential of tissue engineering and regenerative medicine, biomaterials should allow for the independent control of these features. This study investigates a hydrogel system based on thiol-modified hyaluronic acid (HA-S) and polyethylene glycol diacrylate (PEGDA). The mechanical properties of HAS-PEGDA are dictated by two cytocompatible crosslinking reactions that occur at distinct time points: a rapid, Michael-type nucleophilic addition reaction between HA-thiols and PEG-acrylates and a prolonged maturation of disulfide crosslinks from remaining thiols. It is hypothesized that these reactions would enable the independent tuning of the mechanical and bioactive features of HAS-PEGDA. Rheological studies confirmed that initial gelation reached completion by 1 day, at which point the shear modulus was proportional to the concentration of PEGDA. Over time, the shear modulus evolved dramatically, and final stiffness depended on the availability of HA-thiols. The addition of PEG-monoacrylate (PEGMA) after the initial gelation occupied a percentage of remaining thiols to prevent disulfide crosslinking, decreasing the steady-state stiffness in a dose-dependent manner. A fraction of the PEGMA was then replaced with acrylated peptide ligands to introduce specific bioactivity to the otherwise non-cell-adhesive network. The degree of latent stiffening was controlled by the total amount of peptide-PEGMA, while adhesivity was tuned with the balance of bioactive and inactive peptides. The functional effects of the tunable mechanical and bioadhesive ligand properties were confirmed with assays of cell adhesion and morphology.