Mechanical behavior of full-thickness burn human skin is rate-independent.

Skin tissue is recognized to exhibit rate-dependent mechanical behavior under various loading conditions. Here, we report that the full-thickness burn human skin exhibits rate-independent behavior under uniaxial tensile loading conditions. Mechanical properties, namely, ultimate tensile stress, ultimate tensile strain, and toughness, and parameters of Veronda-Westmann hyperelastic material law were assessed via uniaxial tensile tests. Univariate hypothesis testing yielded no significant difference (p > 0.01) in the distributions of these properties for skin samples loaded at three different rates of 0.3 mm/s, 2 mm/s, and 8 mm/s. Multivariate multiclass classification, employing a logistic regression model, failed to effectively discriminate samples loaded at the aforementioned rates, with a classification accuracy of only 40%. The median values for ultimate tensile stress, ultimate tensile strain, and toughness are computed as 1.73 MPa, 1.69, and 1.38 MPa, respectively. The findings of this study hold considerable significance for the refinement of burn care training protocols and treatment planning, shedding new light on the unique, rate-independent behavior of burn skin.

Characterizing the Suture Pullout Force for Human Small Bowel.

Performing a small bowel anastomosis, or reconnecting small bowel segments, remains a core competency and critical step for the successful surgical management of numerous bowel and urinary conditions. As surgical education and technology moves toward improving patient outcomes through automation and increasing training opportunities, a detailed characterization of the interventional biomechanical properties of the human bowel is important. This is especially true due to the prevalence of anastomotic leakage as a frequent (3.02%) postoperative complication of small bowel anastomoses. This study aims to characterize the forces required for a suture to tear through human small bowel (suture pullout force, SPOF), while analyzing how these forces are affected by tissue orientation, suture material, suture size, and donor demographics. 803 tests were performed on 35 human small bowel specimens. A uni-axial test frame was used to tension sutures looped through 10 × 20 mm rectangular bowel samples to tissue failure. The mean SPOF of the small bowel was 4.62±1.40 N. We found no significant effect of tissue orientation (p = 0.083), suture material (p = 0.681), suture size (p = 0.131), age (p = 0.158), sex (p = .083), or body mass index (BMI) (p = 0.100) on SPOF. To our knowledge, this is the first study reporting human small bowel SPOF. Little research has been published about procedure-specific data on human small bowel. Filling this gap in research will inform the design of more accurate human bowel synthetic models and provide an accurate baseline for training and clinical applications.

Characterization of Puncture Forces of the Human Trachea and Cricothyroid Membrane.

Accurate human tissue biomechanical data represents a critical knowledge gap that will help facilitate the advancement of new medical devices, patient-specific predictive models, and training simulators. Tissues related to the human airway are a top priority, as airway medical procedures are common and critical. Placement of a surgical airway, though less common, is often done in an emergent (cricothyrotomy) or urgent (tracheotomy) fashion. This study is the first to report relevant puncture force data for the human cricothyroid membrane and tracheal annular ligaments. Puncture forces of the cricothyroid membrane and tracheal annular ligaments were collected from 39 and 42 excised human donor tracheas, respectively, with a mechanized load frame holding various surgical tools. The average puncture force of the cricothyroid membrane using an 11 blade scalpel was 1.01 ± 0.36 N, and the average puncture force of the tracheal annular ligaments using a 16 gauge needle was 0.98 ± 0.34 N. This data can be used to inform medical device and airway training simulator development as puncture data of these anatomies has not been previously reported.

Thermally damaged porcine skin is not a surrogate mechanical model of human skin.

Porcine skin is considered a de facto surrogate for human skin. However, this study shows that the mechanical characteristics of full thickness burned human skin are different from those of porcine skin. The study relies on five mechanical properties obtained from uniaxial tensile tests at loading rates relevant to surgery: two parameters of the Veronda-Westmann hyperelastic material model, ultimate tensile stress, ultimate tensile strain, and toughness of the skin samples. Univariate statistical analyses show that human and porcine skin properties are dissimilar (p < 0.01) for each loading rate. Multivariate classification involving the five mechanical properties using logistic regression can successfully separate the two skin types with a classification accuracy exceeding 95% for each loading rate individually as well as combined. The findings of this study are expected to guide the development of effective training protocols and high-fidelity simulators to train burn care providers.

Monolithic 3D printing of embeddable and highly stretchable strain sensors using conductive ionogels.

Medical training simulations that utilize 3D-printed, patient-specific tissue models improve practitioner and patient understanding of individualized procedures and capacitate pre-operative, patient-specific rehearsals. The impact of these novel constructs in medical training and pre-procedure rehearsals has been limited, however, by the lack of effectively embedded sensors that detect the location, direction, and amplitude of strains applied by the practitioner on the simulated structures. The monolithic fabrication of strain sensors embedded into lifelike tissue models with customizable orientation and placement could address this limitation. The demonstration of 3D printing of an ionogel as a stretchable, piezoresistive strain sensor embedded in an elastomer is presented as a proof-of-concept of this integrated fabrication for the first time. The significant hysteresis and drift inherent to solid-phase piezoresistive composites and the dimensional instability of low-hysteresis piezoresistive liquids inspired the adoption of a 3D-printable piezoresistive ionogel composed of reduced graphene oxide and an ionic liquid. The shear-thinning rheology of the ionogel obviates the need to fabricate additional structures that define or contain the geometry of the sensing channel. Sensors are printed on and subsequently encapsulated in polydimethylsiloxane (PDMS), a thermoset elastomer commonly used for analog tissue models, to demonstrate seamless fabrication. Strain sensors demonstrate geometry- and strain-dependent gauge factors of 0.54-2.41, a high dynamic strain range of 350% that surpasses the failure strain of most dermal and viscus tissue, low hysteresis (<3.5% degree of hysteresis up to 300% strain) and baseline drift, a single-value response, and excellent fatigue stability (5000 stretching cycles). In addition, we fabricate sensors with stencil-printed silver/PDMS electrodes in place of wires to highlight the potential of seamless integration with printed electrodes. The compositional tunability of ionic liquid/graphene-based composites and the shear-thinning rheology of this class of conductive gels endows an expansive combination of customized sensor geometry and performance that can be tailored to patient-specific, high-fidelity, monolithically fabricated tissue models.

Strong stimulation triggers full fusion exocytosis and very slow endocytosis of the small dense core granules in carotid glomus cells.

Chemosensory glomus cells of the carotid bodies release transmitters, including ATP and dopamine mainly via the exocytosis of small dense core granules (SDCGs, vesicular diameter of ∼100 nm). Using carbon-fiber amperometry, we showed previously that with a modest uniform elevation in cytosolic Ca2+ concentration ([Ca2+]i of ∼0.5 µM), SDCGs of rat glomus cells predominantly underwent a "kiss-and-run" mode of exocytosis. Here, we examined whether a larger [Ca2+]i rise influenced the mode of exocytosis. Activation of voltage-gated Ca2+ channels by a train of voltage-clamped depolarizations which elevated [Ca2+]i to ∼1.6 µM increased the cell membrane capacitance by ∼2.5%. At 30 s after such a stimulus, only 5% of the added membrane was retrieved. Flash photolysis of caged-Ca2+ (which elevated [Ca2+]i to ∼16 µM) increased cell membrane capacitance by ∼13%, and only ∼30% of the added membrane was retrieved at 30 s after the UV flash. When exocytosis and endocytosis were monitored using the two-photon excitation and extracellular polar tracer (TEP) imaging of FM1-43 fluorescence in conjunction with photolysis of caged Ca2+, almost uniform exocytosis was detected over the cell's entire surface and it was followed by slow endocytosis. Immunocytochemistry showed that the cytoplasmic densities of dynamin I, II and clathrin (key proteins that mediate endocytosis) in glomus cells were less than half of those in adrenal chromaffin cells, suggesting that a lower expression of endocytotic machinery may underlie the slow endocytosis in glomus cells. An analysis of the relative change in the signals from two fluorescent dyes that simultaneously monitored the addition of vesicular volume and plasma membrane surface area, suggested that with an intense stimulus, SDCGs of glomus cells underwent full fusion without any significant "compound" exocytosis. Therefore, during a severe hypoxic challenge, glomus granules undergo full fusion for a more complete release of transmitters.

High-throughput and high-yield fabrication of uniaxially-aligned chitosan-based nanofibers by centrifugal electrospinning.

The inability to produce large quantities of nanofibers has been a primary obstacle in advancement and commercialization of electrospinning technologies, especially when aligned nanofibers are desired. Here, we present a high-throughput centrifugal electrospinning (HTP-CES) system capable of producing a large number of highly-aligned nanofiber samples with high-yield and tunable diameters. The versatility of the design was revealed when bead-less nanofibers were produced from copolymer chitosan/polycaprolactone (C-PCL) solutions despite variations in polymer blend composition or spinneret needle gauge. Compared to conventional electrospinning techniques, fibers spun with the HTP-CES not only exhibited superior alignment, but also better diameter uniformity. Nanofiber alignment was quantified using Fast Fourier Transform (FFT) analysis. In addition, a concave correlation between the needle diameter and resultant fiber diameter was identified. This system can be easily scaled up for industrial production of highly-aligned nanofibers with tunable diameters that can potentially meet the requirements for various engineering and biomedical applications.

Effects of convection-enhanced delivery of bevacizumab on survival of glioma-bearing animals.

OBJECT Bevacizumab (Avastin), an antibody to vascular endothelial growth factor (VEGF), alone or in combination with irinotecan (Camptosar [CPT-11]), is a promising treatment for recurrent glioblastoma. However, the intravenous (IV) administration of bevacizumab produces a number of systemic side effects, and the increase in survival it provides for patients with recurrent glioblastoma is still only a few months. Because bevacizumab is an antibody against VEGF, which is secreted into the extracellular milieu by glioma cells, the authors hypothesized that direct chronic intratumoral delivery techniques (i.e., convection-enhanced delivery [CED]) can be more effective than IV administration. To test this hypothesis, the authors compared outcomes for these routes of bevacizumab application with respect to animal survival, microvessel density (MVD), and inflammatory cell distribution. METHODS Two human glioma cell lines, U87 and U251, were used as sources of intracranial tumor cells. The glioma cell lines were implanted into the brains of mice in an orthotopic xenograft mouse tumor model. After 7 days, the mice were treated with one of the following: 1) vehicle, 2) CED bevacizumab, 3) IV bevacizumab, 4) intraperitoneal (IP) irinotecan, 5) CED bevacizumab plus IP irinotecan, or 6) IV bevacizumab plus IP irinotecan. Alzet micro-osmotic pumps were used to introduce bevacizumab directly into the tumor. Survival was monitored. Excised tumor tissue samples were immunostained to measure MVD and inflammatory cell and growth factor levels. RESULTS The results demonstrate that mice treated with CED of bevacizumab alone or in combination with irinotecan survived longer than those treated systemically; CED-treated animals survived 30% longer than IV-treated animals. In combination studies, CED bevacizumab plus CPT-11 increased survival by more than 90%, whereas IV bevacizumab plus CPT-11 increased survival by 40%. Furthermore, CED bevacizumab-treated tissues exhibited decreased MVD compared with that of IV-treated tissues. In additional studies, the infiltration of macrophages and dendritic cells into CED-treated animals were increased compared with those in IV-treated animals, suggesting a highly active inflammatory response taking place in CED-treated mice. CONCLUSIONS The administration of bevacizumab via CED increases survival over that of treatment with IV bevacizumab. Thus, CED of bevacizumab alone or in combination with chemotherapy can be an effective protocol for treating gliomas.

Argonaute-2 promotes miR-18a entry in human brain endothelial cells.

BACKGROUND: Cerebral arteriovenous malformation (AVM) is a vascular disease exhibiting abnormal blood vessel morphology and function. miR-18a ameliorates the abnormal characteristics of AVM-derived brain endothelial cells (AVM-BEC) without the use of transfection reagents. Hence, our aim was to identify the mechanisms by which miR-18a is internalized by AVM-BEC. Since AVM-BEC overexpress RNA-binding protein Argonaute-2 (Ago-2) we explored the clinical potential of Ago-2 as a systemic miRNA carrier. METHODS AND RESULTS: Primary cultures of AVM-BEC were isolated from surgical specimens and tested for endogenous miR-18a levels using qPCR. Conditioned media (CM) was derived from AVM-BEC cultures (AVM-BEC-CM). AVM-BEC-CM significantly enhanced miR-18a internalization. Ago-2 was detected using western blotting and immunostaining techniques. Ago-2 was highly expressed in AVM-BEC; and siAgo-2 decreased miR-18a entry into brain-derived endothelial cells. Only brain-derived endothelial cells were responsive to the Ago-2/miR-18a complex and not other cell types tested. Secreted products (eg, thrombospondin-1 [TSP-1]) were tested using ELISA. Brain endothelial cells treated with the Ago-2/miR-18a complex in vitro increased TSP-1 secretion. In the in vivo angiogenesis glioma model, animals were treated with miR-18a in combination with Ago-2. Plasma was obtained and tested for TSP-1 and vascular endothelial growth factor (VEGF)-A. In this angiogenesis model, the Ago-2/miR-18a complex caused a significant increase in TSP-1 and decrease in VEGF-A secretion in the plasma. CONCLUSIONS: Ago-2 facilitates miR-18a entry into brain endothelial cells in vitro and in vivo. This study highlights the clinical potential of Ago-2 as a miRNA delivery platform for the treatment of brain vascular diseases.