Effects of internal electrode cooling on irreversible electroporation using a perfused organ model.

PURPOSE: This study evaluates the effects of active electrode cooling, via internal fluid circulation, on the irreversible electroporation (IRE) lesion, deployed electric current and temperature changes using a perfused porcine liver model. MATERIALS AND METHODS: A bipolar electrode delivered IRE electric pulses with or without activation of internal cooling to nine porcine mechanically perfused livers. Pulse schemes included a constant voltage, and a preconditioned delivery combined with an arc-mitigation algorithm. After treatment, organs were dissected, and treatment zones were stained using triphenyl-tetrazolium chloride (TTC) to demonstrate viability. RESULTS: Thirty-nine treatments were performed with an internally cooled applicator and 21 with a non-cooled applicator. For the constant voltage scenario, the average final electrical current measured was 26.37 and 29.20 A for the cooled and uncooled electrodes respectively ([Formula: see text]). The average final temperature measured was 33.01 and 42.43 °C for the cooled and uncooled electrodes respectively ([Formula: see text]). The average measured ablations (fixed lesion) were 3.88-by-2.08 cm and 3.86-by-2.12 cm for the cooled and uncooled electrode respectively ([Formula: see text], [Formula: see text]). Similarly, the preconditioned/arc-mitigation scenario yielded an average final electrical current measurement of a 41.07 and 47.20 A for the cooled and uncooled electrodes respectively ([Formula: see text]). The average final temperature measured was 34.93 and 44.90 °C for the cooled and uncooled electrodes respectively ([Formula: see text]). The average measured ablations (fixed lesion) were 3.67-by-2.27 cm and 3.58-by-2.09 cm for the cooled and uncooled applicators ([Formula: see text]). CONCLUSIONS: The internally-cooled bipolar applicator offers advantages that could improve clinical outcomes. Thermally mitigating internal perfusion technology reduced tissue temperatures and electric current while maintaining similar lesion sizes.

3D printed conformal microfluidics for isolation and profiling of biomarkers from whole organs.

The ability to interface microfluidic devices with native complex biological architectures, such as whole organs, has the potential to shift the paradigm for the study and analysis of biological tissue. Here, we show 3D printing can be used to fabricate bio-inspired conformal microfluidic devices that directly interface with the surface of whole organs. Structured-light scanning techniques enabled the 3D topographical matching of microfluidic device geometry to porcine kidney anatomy. Our studies show molecular species are spontaneously transferred from the organ cortex to the conformal microfluidic device in the presence of fluid flow through the organ-conforming microchannel. Large animal studies using porcine kidneys (n = 32 organs) revealed the profile of molecular species in the organ-conforming microfluidic stream was dependent on the organ preservation conditions. Enzyme-linked immunosorbent assay (ELISA) studies revealed conformal microfluidic devices isolate clinically relevant metabolic and pathophysiological biomarkers from whole organs, including heat shock protein 70 (HSP-70) and kidney injury molecule-1 (KIM-1), which were detected in the microfluidic device as high as 409 and 12 pg mL-1, respectively. Overall, these results show conformal microfluidic devices enable a novel minimally invasive 'microfluidic biopsy' technique for isolation and profiling of biomarkers from whole organs within a clinically relevant interval. This achievement could shift the paradigm for whole organ preservation and assessment, thereby helping to relieve the organ shortage crisis through increased availability and quality of donor organs. Ultimately, this work provides a major advance in microfluidics through the design and manufacturing of organ-conforming microfluidic devices and a novel technique for microfluidic-based analysis of whole organs.

The Development of a Thin-Filmed Noninvasive Tissue Perfusion Sensor to Quantify Capillary Pressure Occlusion of Explanted Organs.

A new thin-filmed perfusion sensor was developed using a heat flux gauge, thin-film thermocouple, and a heating element. This sensor, termed "CHFT+," is an enhancement of the previously established combined heat flux-temperature (CHFT) sensor technology predominately used to quantify the severity of burns [1]. The CHFT+ sensor was uniquely designed to measure tissue perfusion on explanted organs destined for transplantation, but could be functionalized and used in a wide variety of other biomedical applications. Exploiting the thin and semiflexible nature of the new CHFT+ sensor assembly, perfusion measurements can be made from the underside of the organ-providing a quantitative indirect measure of capillary pressure occlusion. Results from a live tissue test demonstrated, for the first time, the effects of pressure occlusion on an explanted porcine kidney. CHFT+ sensors were placed on top of and underneath 18 kidneys to measure and compare perfusion at perfusate temperatures of 5 and 20 °C. The data collected show a greater perfusion on the topside than the underside of the specimen for the length of the experiment. This indicates that the pressure occlusion is truly affecting the perfusion, and, thus, the overall preservation of explanted organs. Moreover, the results demonstrate the effect of preservation temperature on the tissue vasculature. Focusing on the topside perfusion only, the 20 °C perfusion was greater than the 5 °C perfusion, likely due to the vasoconstrictive response at the lower perfusion temperatures.

Assessment of the accuracy of computed tomography for measurement of normal equine pituitary glands.

OBJECTIVE: To describe the anatomic features of the pituitary gland region in horses via computed tomography (CT) and determine the accuracy of CT for estimating normal equine pituitary gland dimensions. ANIMALS: 25 adult horses with no clinical signs of pituitary disease. PROCEDURE: Transverse CT images and gross transverse tissue sections were compared in 2 horses. Contrast-enhanced CT of the pituitary gland region was performed postmortem in 23 horses with 4 slice thickness and interval settings (10-mm contiguous or overlapping slices and 4-mm contiguous or overlapping slices). Gross and CT estimates of pituitary gland dimensions were compared via ANOVA. Accuracy of CT estimates was calculated with gross pituitary gland measurements as the known value. RESULTS: Pituitary glands were located between the temporomandibular joints and had contrast enhancement. Mean gross dimensions were length, 2.11 cm; width, 2.16 cm; height, 0.98 cm; and volume, 2.66 cm3. Gross measurements and CT estimates of pituitary gland length from 10-mm contiguous and overlapping slices did not differ. Gross measurements and CT estimates of pituitary gland width from 4-mm contiguous and overlapping slices did not differ. Estimates of height and volume from all CT techniques differed from gross measurements. Accuracies for CT estimates were length, 88 to 99%; width, 81 to 92%; height, 58 to 71%; and volume, 43 to 55%. CONCLUSIONS AND CLINICAL RELEVANCE: Accuracy of estimates of pituitary gland dimension in horses varied with CT scanning technique; via CT estimates of length and width of glands were more accurate than estimates of height or volume.