Electrical stimulation affects the differentiation of transplanted regionally specific human spinal neural progenitor cells (sNPCs) after chronic spinal cord injury.

BACKGROUND: There are currently no effective clinical therapies to ameliorate the loss of function that occurs after spinal cord injury. Electrical stimulation of the rat spinal cord through the rat tail has previously been described by our laboratory. We propose combinatorial treatment with human induced pluripotent stem cell-derived spinal neural progenitor cells (sNPCs) along with tail nerve electrical stimulation (TANES). The purpose of this study was to examine the influence of TANES on the differentiation of sNPCs with the hypothesis that the addition of TANES would affect incorporation of sNPCs into the injured spinal cord, which is our ultimate goal. METHODS: Chronically injured athymic nude rats were allocated to one of three treatment groups: injury only, sNPC only, or sNPC + TANES. Rats were sacrificed at 16 weeks post-transplantation, and tissue was processed and analyzed utilizing standard histological and tissue clearing techniques. Functional testing was performed. All quantitative data were presented as mean ± standard error of the mean. Statistics were conducted using GraphPad Prism. RESULTS: We found that sNPCs were multi-potent and retained the ability to differentiate into mainly neurons or oligodendrocytes after this transplantation paradigm. The addition of TANES resulted in more transplanted cells differentiating into oligodendrocytes compared with no TANES treatment, and more myelin was found. TANES not only promoted significantly higher numbers of sNPCs migrating away from the site of injection but also influenced long-distance axonal/dendritic projections especially in the rostral direction. Further, we observed localization of synaptophysin on SC121-positive cells, suggesting integration with host or surrounding neurons, and this finding was enhanced when TANES was applied. Also, rats that were transplanted with sNPCs in combination with TANES resulted in an increase in serotonergic fibers in the lumbar region. This suggests that TANES contributes to integration of sNPCs, as well as activity-dependent oligodendrocyte and myelin remodeling of the chronically injured spinal cord. CONCLUSIONS: Together, the data suggest that the added electrical stimulation promoted cellular integration and influenced the fate of human induced pluripotent stem cell-derived sNPCs transplanted into the injured spinal cord.

3D Printed Skin-Interfaced UV-Visible Hybrid Photodetectors.

Photodetectors that are intimately interfaced with human skin and measure real-time optical irradiance are appealing in the medical profiling of photosensitive diseases. Developing compliant devices for this purpose requires the fabrication of photodetectors with ultraviolet (UV)-enhanced broadband photoresponse and high mechanical flexibility, to ensure precise irradiance measurements across the spectral band critical to dermatological health when directly applied onto curved skin surfaces. Here, a fully 3D printed flexible UV-visible photodetector array is reported that incorporates a hybrid organic-inorganic material system and is integrated with a custom-built portable console to continuously monitor broadband irradiance in-situ. The active materials are formulated by doping polymeric photoactive materials with zinc oxide nanoparticles in order to improve the UV photoresponse and trigger a photomultiplication (PM) effect. The ability of a stand-alone skin-interfaced light intensity monitoring system to detect natural irradiance within the wavelength range of 310-650 nm for nearly 24 h is demonstrated.

Effects of solvent osmolarity and viscosity on cartilage energy dissipation under high-frequency loading.

Articular cartilage is a spatially heterogeneous, dissipative biological hydrogel with a high fluid volume fraction. Although energy dissipation is important in the context of delaying cartilage damage, the dynamic behavior of articular cartilage equilibrated in media of varied osmolarity and viscosity is not widely understood. This study investigated the mechanical behaviors of cartilage when equilibrated to media of varying osmolarity and viscosity. Dynamic moduli and phase shift were measured at both low (1 Hz) and high (75-300 Hz) frequency, with cartilage samples compressed to varied offset strain levels. Increasing solution osmolarity and viscosity both independently resulted in larger energy dissipation and decreased dynamic modulus of cartilage at both low and high frequency. Mechanical property alterations induced by varying osmolarity are likely due to the change in permeability and fluid volume fraction within the tissue. The effects of solution viscosity are likely due to frictional interactions at the solid-fluid interface, affecting energy dissipation. These findings highlight the significance of interstitial fluid on the energy dissipation capabilities of the tissue, which can influence the onset of cartilage damage.

Microindentation Technique to Create Localized Cartilage Microfractures.

Articular cartilage is a multiphasic, anisotropic, and heterogeneous material. Although cartilage possesses excellent mechanical and biological properties, it can undergo mechanical damage, resulting in osteoarthritis. Thus, it is important to understand the microscale failure behavior of cartilage in both basic science and clinical contexts. Determining cartilage failure behavior and mechanisms provides insight for improving treatment strategies to delay osteoarthritis initiation or progression and can also enhance the value of cartilage as bioinspiration for material fabrication. To investigate microscale failure behavior, we developed a protocol to initiate fractures by applying a microindentation technique using a well-defined tip geometry that creates localized cracks across a range of loading rates. The protocol includes extracting the tissue from the joint, preparing samples, and microfracture. Various aspects of the experiment, such as loading profile and solvent, can be adjusted to mimic physiological or pathological conditions and thereby further clarify phenomena underlying articular cartilage failure. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Harvesting and dissection of the joint surfaces Basic Protocol 2: Preparation of samples for microindentation and fatigue testing Basic Protocol 3: Microfracture using microindentation Basic Protocol 4: Crack propagation under cyclic loading.

Glycosaminoglycan depletion increases energy dissipation in articular cartilage under high-frequency loading.

High-frequency material behavior of cartilage at macroscopic lengths is not widely understood, despite a wide range of frequencies and contact lengths experienced in vivo. For example, cartilage at different stages of matrix integrity can experience high-frequency loading during traumatic impact, making high-frequency behavior relevant in the context of structural failure. Therefore, this study examined macroscopic dissipative and mechanical responses of intact and glycosaminoglycan (GAG)-depleted cartilage under previously unexplored high-frequency loading. These dynamic responses were complemented with the evaluation of quasi-static responses. A custom dynamic mechanical analyzer was used to obtain dynamic behavior, and stress relaxation testing was performed to obtain quasi-static behavior. Under high-frequency loading, cartilage energy dissipation increased with GAG depletion and decreased with strain; dynamic modulus exhibited opposite trends. Similarly, under quasi-static loading, equilibrium modulus and relaxation time of cartilage decreased with GAG depletion. The increased energy dissipation after GAG depletion under high-frequency loading was likely due to increased viscoelastic dissipation. These findings broaden our understanding of fundamental properties of cartilage as a function of solid matrix integrity in an unprecedented loading regime. They also provide a foundation for analyzing energy dissipation associated with cartilage failure induced by traumatic impact.

Rate-dependent adhesion of cartilage and its relation to relaxation mechanisms.

Cartilage adhesion has been found to play an important role in friction responses in the boundary lubrication regime, but its underlying mechanisms have only been partially understood. This study investigates the rate dependence of adhesion from pre-to post-relaxation timescales of cartilage and its possible relation to relaxation responses of the tissue. Adhesion tests on cartilage were performed to obtain rate-dependent cartilage adhesion from relaxed to unrelaxed states and corresponding relaxation responses. The rate dependence of cartilage adhesion was analyzed based on experimental relaxation responses. Cartilage adhesion increased about 20 times from relaxed to unrelaxed states. This rate-dependent enhancement correlated well with the load relaxation responses in a characteristic time domain. These experimental results indicated that the degree of recovery (or relaxation) in the vicinity of contact during unloading governed the rate dependence of cartilage adhesion. In addition, the experimentally measured enhancement of adhesion was interpreted with the aid of computationally and analytically predicted adhesion trends in viscoelastic, poroviscoelastic, and cohesive contact models. Agreement between the experimental and predicted trends implied that the enhancement of cartilage adhesion originated from complex combinations of interfacial peeling and negative fluid pressure generated within the contact area during unloading. These findings enhance the current understanding of rate-dependent adhesion mechanisms explored within short time scales and thus could provide new insight into friction responses and stick-induced damage in cartilage.

Rate-dependent crack nucleation in cartilage under microindentation.

This study investigates rate-dependent crack nucleation in cartilage under microindentation using a poroviscoelastic framework and nano/microscopic images. Localized crack failure was induced at known locations and at different loading rates via microindentation with an axisymmetric sphero-conical indenter. Finite element (FE) modeling was used to reproduce results of microindentation tests within a poroviscoelastic framework. Scanning electron microscopy (SEM) was used to examine nano- and microscale structural features of crack surfaces. Microindentation results showed rate-dependent crack nucleation in cartilage. In particular, critical total work required for crack nucleation was larger at the slow loading rate compared to the fast loading rate. FE results suggested that viscoelastic relaxation of cartilage was a major contributor to the rate dependency and that tensile stresses localized at the indenter tip was a governing factor in crack nucleation. SEM images combined with microindentation and FE results suggested that the solid matrix in the vicinity of the tip experienced relatively large relaxation and kinematic fiber rearrangement at the slow loading rate in comparison to the fast loading rate. These findings extend current understanding of rate-dependent failure mechanisms in cartilage.

Effect of relaxation-dependent adhesion on pre-sliding response of cartilage.

Possible links between adhesive properties and the pre-sliding (static) friction response of cartilage are not fully understood in the literature. The aims of this study are to investigate the relation between adhesion and relaxation time in articular cartilage, and the effect of relaxation-dependent adhesion on the pre-sliding response of cartilage. Adhesion tests were performed to evaluate the work of adhesion of cartilage at different relaxation times. Friction tests were conducted to identify the pre-sliding friction response of cartilage at relaxation times corresponding to adhesion tests. The pre-sliding friction response of cartilage was systematically linked to the work of adhesion and contact conditions by a slip-based failure model. It was found that the work of adhesion increases with relaxation time. Also, the work of adhesion is linearly correlated to the resistance to slip-based failure. In addition, as the work of adhesion increases, the adhered (stick) area at the moment of failure increases, and the propagation rate of the annular slip (crack) area towards its centre increases. These findings offer a mechanistic explanation of the pre-sliding friction behaviour and stick-slip response of soft hydrated interfaces such as articular cartilage and hydrogels. In addition, the linear correlation between adhesion and threshold to slip-based failure enables estimation of the adhesive strength of such interfaces directly from the pre-sliding friction response (e.g. shear wave elastography).

Uncoupled poroelastic and intrinsic viscoelastic dissipation in cartilage.

This paper studies uncoupled poroelastic (flow-dependent) and intrinsic viscoelastic (flow-independent) energy dissipation mechanisms via their dependence on characteristic lengths to understand the root of cartilage's broadband dissipation behavior. Phase shift and dynamic modulus were measured from dynamic microindentation tests conducted on hydrated cartilage at different contact radii, as well as on dehydrated cartilage. Cartilage weight and thickness were recorded during dehydration. Phase shifts revealed poroelastic- and viscoelastic-dominant dissipation regimes in hydrated cartilage. Specifically, phase shift at a relatively small radius was governed by poroviscoelasticity, while phase shift at a relatively large radius was dominantly governed by intrinsic viscoelasticity. The uncoupled dissipation mechanisms demonstrated that intrinsic viscoelastic dissipation provided sustained broadband dissipation for all length scales, and additional poroelastic dissipation increased total dissipation at small length scales. Dehydration decreased intrinsic viscoelastic dissipation of cartilage. The findings demonstrated a possibility to measure poroelastic and intrinsic viscoelastic properties of cartilage at similar microscale lengths. Also they encouraged development of broadband cartilage like-dampers and provided important design parameters to maximize their performance.

Fabrication of tungsten probe for hard tapping operation in atomic force microscopy.

We propose a method of producing a tungsten probe with high stiffness for atomic force microscopy (AFM) in order to acquire enhanced phase contrast images and efficiently perform lithography. A tungsten probe with a tip radius between 20nm and 50nm was fabricated using electrochemical etching optimized by applying pulse waves at different voltages. The spring constant of the tungsten probe was determined by finite element analysis (FEA), and its applicability as an AFM probe was evaluated by obtaining topography and phase contrast images of a Si wafer sample partly coated with Au. Enhanced hard tapping performance of the tungsten probe compared with a commercial Si probe was confirmed by conducting hard tapping tests at five different oscillation amplitudes on single layer graphene grown by chemical vapor deposition (CVD). To analyze the damaged graphene sample, the test areas were investigated using tip-enhanced Raman spectroscopy (TERS). The test results demonstrate that the tungsten probe with high stiffness was capable of inducing sufficient elastic and plastic deformation to enable obtaining enhanced phase contrast images and performing lithography, respectively.

High mechanical and tribological stability of an elastic ultrathin overcoating layer for flexible silver nanowire films.

A novel, nanoscale, thickness-controlled, elastic graphene oxide-polydiallyldimethylammonium chloride (GO-PDDA) film using a layer-by-layer technique on silver nanowires and a flexible substrate is reported. Micro- and nanoscale wear and flexibility depending on the thickness and/or elastic nature of the overcoating layer demonstrate high mechanical stability with the PDDA inserted overcoating layer.