High-intensity focused ultrasound ablation of muscle in an anticoagulated swine model.

INTRODUCTION: Image-guided non-invasive high-intensity focused ultrasound (HIFU) has been gaining recognition in treating musculoskeletal tumors and desmoids. However, there is no consensus on the appropriate perioperative management for patients on ongoing anticoagulation who undergo HIFU ablation. MATERIAL AND METHODS: Image-guided HIFU treatment was performed in swine on an ongoing oral anticoagulation protocol (N = 5) in two treatment sessions seven days apart. On day one, a total of twenty locations were ablated, and on day eight, ten more muscle ablations were performed, and the animals were euthanized. Imaging, clinical examination, and histopathology were performed to investigate treated tissue for bleeding. RESULTS: Imaging, clinical examination, and histopathology revealed either no bleeding or, in some samples, only small scattered cavities (0.2-2 mm in diameter) filled with blood. CONCLUSION: Noninvasive HIFU ablation of muscle may not require a coagulation profile within normal limits.

The Mouse Round-window Approach for Ototoxic Agent Delivery: A Rapid and Reliable Technique for Inducing Cochlear Cell Degeneration.

Investigators have utilized a wide array of animal models and investigative techniques to study the mammalian auditory system. Much of the basic research involving the cochlea and its associated neural pathways entails exposure of model cochleae to a variety of ototoxic agents. This allows investigators to study the effects of targeted damage to cochlear structures, and in some cases, the self-repair or regeneration of those structures. Various techniques exist for delivery of ototoxic agents to the cochlea. When selecting a particular technique, investigators must consider a number of factors, including the induction of inadvertent systemic toxicity, the amount of cochlear damage produced by the surgical procedure itself, the type of lesion desired, animal survivability, and reproducibility/reliability of results. Currently established techniques include parenteral injection, intra-peritoneal injection, trans-tympanic injection, endolymphatic sac injection, and cochleostomy with perilymphatic perfusion. Each of these methods has been successfully utilized and is well described in the literature; yet, each has various shortcomings. Here, we present a technique for topical application of ototoxic agents directly to the round window niche. This technique is non-invasive to inner ear structures, produces rapid onset of reliably targeted lesions, avoids systemic toxicity, and allows for an intra-animal control (the contra-lateral ear). Results stemming from this approach have helped deeper understanding of auditory pathophysiology, cochlear cell degeneration, and regenerative capacity in response to an acute injury. Future investigations may use this method to conduct interventional studies involving gene therapy and stem cell transplantation to combat hearing loss.

Feasibility Study on MR-Guided High-Intensity Focused Ultrasound Ablation of Sciatic Nerve in a Swine Model: Preliminary Results.

INTRODUCTION: Spastic patients often seek neurolysis, the permanent destruction of the sciatic nerve, for better pain management. MRI-guided high-intensity focused ultrasound (MRgHIFU) may serve as a noninvasive alternative to the prevailing, more intrusive techniques. This in vivo acute study is aimed at performing sciatic nerve neurolysis using a clinical MRgHIFU system. METHODS: The HIFU ablation of sciatic nerves was performed in swine (n = 5) using a HIFU system integrated with a 3 T MRI scanner. Acute lesions were confirmed using T1-weighted contrast-enhanced (CE) MRI and histopathology using hematoxylin and eosin staining. The animals were euthanized immediately following post-ablation imaging. RESULTS: Reddening and mild thickening of the nerve and pallor of the adjacent muscle were seen in all animals. The HIFU-treated sections of the nerves displayed nuclear pyknosis of Schwann cells, vascular hyperemia, perineural edema, hyalinization of the collagenous stroma of the nerve, myelin sheet swelling, and loss of axons. Ablations were visible on CE MRI. Non-perfused volume of the lesions (5.8-64.6 cc) linearly correlated with estimated lethal thermal dose volume (4.7-34.2 cc). Skin burn adjacent to the largest ablated zone was observed in the first animal. Bilateral treatment time ranged from 55 to 138 min, and preparation time required 2 h on average. CONCLUSION: The acute pilot study in swine demonstrated the feasibility of a noninvasive neurolysis of the sciatic nerve using a clinical MRgHIFU system. Results revealed that acute HIFU nerve lesions were detectable on CE MRI, gross pathology, and histology.

Characterization of bone perfusion by dynamic contrast-enhanced magnetic resonance imaging and positron emission tomography in the Dunkin-Hartley guinea pig model of advanced osteoarthritis.

This study characterizes changes in subchondral bone circulation in OA and examines relationships to bone structure and cartilage degeneration in Dunkin-Hartley guinea pigs. We have used dynamic contrast-enhanced MRI (DCE-MRI) and PET, with pharmacokinetic modeling, to characterize subchondral bone perfusion. Assessments are made of perfusion kinetics and vascular permeability by MRI, and blood volume and flow, and radionuclide incorporation into bone, by PET. These parameters are compared to cartilage lesion severity and bone histomorphometry. Assessments of intraosseous thrombi are made morphologically. Prolonged signal enhancement during the clearance phase of MRI correlated with OA severity and suggested venous stasis. Vascular permeability was not increased indicating that transvascular migration of contrast agent was not responsible for signal enhancement. Intraosseous thrombi were not observed. Decreased perfusion associated with severe OA was confirmed by PET and was associated with reduced radionuclide incorporation and osteoporosis. MRI and PET can be used to characterize kinetic parameters of circulation in OA and correlate them with subchondral bone metabolism of interest to the pathophysiology of OA. The significance of these observations may lie in alterations induced in the expression of cytokines by OA osteoblasts that are related to bone remodeling and cartilage breakdown.

Planning irreversible electroporation in the porcine kidney: are numerical simulations reliable for predicting empiric ablation outcomes?

PURPOSE: Numerical simulations are used for treatment planning in clinical applications of irreversible electroporation (IRE) to determine ablation size and shape. To assess the reliability of simulations for treatment planning, we compared simulation results with empiric outcomes of renal IRE using computed tomography (CT) and histology in an animal model. METHODS: The ablation size and shape for six different IRE parameter sets (70-90 pulses, 2,000-2,700 V, 70-100 µs) for monopolar and bipolar electrodes was simulated using a numerical model. Employing these treatment parameters, 35 CT-guided IRE ablations were created in both kidneys of six pigs and followed up with CT immediately and after 24 h. Histopathology was analyzed from postablation day 1. RESULTS: Ablation zones on CT measured 81 ± 18 % (day 0, p ≤ 0.05) and 115 ± 18 % (day 1, p ≤ 0.09) of the simulated size for monopolar electrodes, and 190 ± 33 % (day 0, p ≤ 0.001) and 234 ± 12 % (day 1, p ≤ 0.0001) for bipolar electrodes. Histopathology indicated smaller ablation zones than simulated (71 ± 41 %, p ≤ 0.047) and measured on CT (47 ± 16 %, p ≤ 0.005) with complete ablation of kidney parenchyma within the central zone and incomplete ablation in the periphery. CONCLUSION: Both numerical simulations for planning renal IRE and CT measurements may overestimate the size of ablation compared to histology, and ablation effects may be incomplete in the periphery.

Feasibility and Safety of Laparoscopic Colon Surgery Performed Under Intravenous Sedation and Local Anesthesia Using Microinvasive (<3 mm) Instruments: An Acute and Survival Study on Porcine Model.

PURPOSE: The purpose of the study was to evaluate the feasibility and safety of performing laparoscopic intestinal surgery using local anesthesia and intravenous sedation with instruments <3 mm in diameter. METHODS: Porcine model with acute (n = 2) and the survival studies (n = 8): all female pigs, weight (median 36.4 kg, range 33.2-38.4 kg). Surgeries were performed using only intravenous sedation with ketamine-midazolam and local anesthetic infiltration at the sites of trocar insertion, with airway protection. CO2 pneumoperitoneum was maintained using pressure of 3 to 5 mm Hg. Commercially available instruments, sizes <3 mm in diameter were used. Surgical steps were as follows: (a) exploration of all quadrants of the abdomen and pelvis, (b) "running" the entire length of small bowel, (c) dissection of bowel attachments to the peritoneal sidewall, and (d) creating a 2.5 cm enterotomy in the colon and suture repair of this defect. RESULTS: All 10 surgeries were completed successfully. Animals tolerated the procedure well, with no requirement of intubation. There were no decrements in vital signs during pneumoperitoneum or surgery. Despite spontaneous respiration movements, all planned surgical maneuvers were feasible. The median length of operations was 74 minutes (range 56-165 minutes). All survival animals had an uneventful recovery; there were no infectious complications, oral intake and bowel function returned within 24 hours. CONCLUSIONS: It appears feasible and safe to perform simple laparoscopic intestinal procedures using instruments <3 mm in diameter and low CO2 insufflation pressure under local anesthesia and intravenous sedation. This methodology holds promise in the development of new approaches to intestinal surgery and disease diagnosis.

MRI-safe robot for targeted transrectal prostate biopsy: animal experiments.

OBJECTIVES: To study the feasibility and safety of using a magnetic resonance imaging (MRI)-safe robot for assisting MRI-guided transrectal needle placement and biopsy in the prostate, using a canine model. To determine the accuracy and precision afforded by the use of the robot while targeting a desired location in the organ. MATERIALS AND METHODS: In a study approved by the Institutional Animal Care and Use Committee, six healthy adult male beagles with prostates of at least 15 × 15 mm in size at the largest transverse section were chosen for the procedure. The probe portion of the robot was placed into the rectum of the dog, images were acquired and image-to-robot registration was performed. Images acquired after placement of the robot were reviewed and a radiologist selected targets for needle placement in the gland. Depending on the size of the prostate, up to a maximum of six needle placements were performed on each dog. After needle placement, robot-assisted core biopsies were performed on four dogs that had larger prostate volumes and extracted cores were analysed for potential diagnostic value. RESULTS: Robot-assisted MRI-guided needle placements were performed to target a total of 30 locations in six dogs, achieving a targeting accuracy of 2.58 mm (mean) and precision of 1.31 mm (SD). All needle placements were successfully completed on the first attempt. The mean time required to select a desired target location in the prostate, align the needle guide to that point, insert the needle and perform the biopsy was ∼ 3 min. For this targeting accuracy study, the inserted needle was also imaged after its placement in the prostate, which took an additional 6-8 min. Signal-to-noise ratio analysis indicated that the presence of the robot within the scanner bore had minimal impact on the quality of the images acquired. Analysis of intact biopsy core samples indicated that the samples contained prostatic tissues, appropriate for making a potential diagnosis. Dogs used in the study did not experience device- or procedure-related complications. CONCLUSIONS: Results from this preclinical pilot animal study suggest that MRI-targeted transrectal biopsies are feasible to perform and this procedure may be safely assisted by an MRI-safe robotic device.

Intraoperative ultrasound and tissue elastography measurements do not predict the size of hepatic microwave ablations.

RATIONALE AND OBJECTIVES: Image-guided thermal ablation is used to treat primary and secondary liver cancers. Evaluating completeness of ablation is difficult with standard intraoperative B-mode ultrasound. This study evaluates the ability of B-mode ultrasound (US) and tissue elastography to adequately measure the extent of ablation compared to pathologic assessment. MATERIALS AND METHODS: An in vivo porcine model was used to compare B-mode ultrasonography and elastography to pathologic assessment of the microwave ablation zone area. In parallel, intraoperative ablations in patients were used to assess the ability of B-mode US and elastographic measures of tissue strain immediately after ablation to predict ablation size, compared to postprocedural computed tomography (CT). RESULTS: In the animal model, ablation zones appeared to decrease in size when monitored with ultrasound over a 10-minute span with both B-mode US and elastography. Both techniques estimated smaller zones than gross pathology, however, the differences did not reach statistical significance. Biopsies from the edges of the ablation zone, as assessed by US, contained viable tissue in 75% of the cases. In the human model, B-mode US and elastography estimated similar ablation sizes; however, they underestimate the final size of the ablation defect as measured on postprocedure CT scan (median area [interquartile range]: CT, 7.3 cm(2) [5.2-9.5] vs. US 3.6 cm(2) [1.7-6.3] and elastography 4.1 cm(2) [1.4-5.1]; P = .005). CONCLUSIONS: Ultrasound and elastography provide an accurate gross estimation of ablation zone size but are unable to predict the degree of cellular injury and significantly underestimate the ultimate size of the ablation.

Novel use of noninvasive high-intensity focused ultrasonography for intercostal nerve neurolysis in a swine model.

BACKGROUND: High-intensity focused ultrasound (HIFU) is a noninvasive thermal ablation technique. High-intensity focused ultrasound has been used in small-animal models to lesion neural tissue selectively. This study aimed to evaluate the efficacy of HIFU in a large-animal model for ablation of nerves similar in size to human nerves. METHODS: Twelve acute magnetic resonance-guided HIFU ablation lesions were created in intercostal nerves in a swine model. In a second pig, as a control, 4 radiofrequency ablation and 4 alcohol lesions were performed on intercostal nerves under ultrasound guidance. Preprocedural and postprocedural magnetic resonance imaging was then performed to evaluate radiologically the lesion size created by HIFU. Animals were euthanized 1 hour postprocedure, and necropsy was performed to collect tissue samples for histopathologic analysis. RESULTS: On gross and histological examination of the intercostal nerve, acute HIFU nerve lesions showed evidence of well-demarcated, acute, focally extensive thermal necrosis. Four intercostal nerves ablated with HIFU were sent for histopathologic analysis, with 2 of 4 lesions showing pathologic damage to the intercostal nerve. Similar results were shown with radiofrequency ablation technique, whereas the intercostal nerves appeared histologically intact with alcohol ablation. CONCLUSIONS: High-intensity focused ultrasound may be used as a noninvasive neurolytic technique in swine. High-intensity focused ultrasound may have potential as a neuroablation technique for patients with chronic and cancer pain.

Irreversible electroporation facilitates gene transfer of a GM-CSF plasmid with a local and systemic response.

BACKGROUND: Electroporation uses an electric field to induce pores in the cell membrane that can transfer macromolecules into target cells. Modulation of electrical parameters leads to irreversible electroporation (IRE), which is being developed for tissue ablation. We sought to evaluate whether the application of IRE may induce a lesser electric field in the periphery where reversible electroporation may occur, facilitating gene transfer of a granulocyte macrophage colony-stimulating factor (GM-CSF) plasmid to produce its biologic response. METHODS: Yorkshire pigs underwent laparotomy, and IRE of the liver was performed during hepatic arterial infusion of 1 or 7 mg of a naked human GM-CSF plasmid. The serum, liver, lymph nodes, and bone marrow were harvested for analysis. RESULTS: Human GM-CSF level rose from undetectable to 131 pg/mL in the serum at 24 hours after IRE and plasmid infusion. The liver demonstrated an ablation zone surrounded by an immune infiltrate that had greater macrophage intensity than when treated with IRE or plasmid infusion alone. This dominance of macrophages was dose dependent. Distant effects of GM-CSF were found in the bone marrow, where proliferating myeloid cells increased from 14% to 25%. CONCLUSION: IRE facilitated gene transfer of the GM-CSF plasmid and brought about a local and systemic biologic response. This technique holds potential for tumor eradication and immunotherapy of residual cancer.

Comparison of simulation-based treatment planning with imaging and pathology outcomes for percutaneous CT-guided irreversible electroporation of the porcine pancreas: a pilot study.

PURPOSE: To investigate the reliability of simulations for planning pancreatic irreversible electroporation (IRE) ablations compared with computed tomography (CT) and pathology outcomes in an animal model. MATERIALS AND METHODS: Simulations were performed varying treatment parameters, including field strength (1.5-2.5 kV/cm), pulse number (70-90 pulses), and pulse length (70-100 µs). Pancreatic IRE was performed in six pigs under CT guidance. Two animals each were sacrificed for histology after 1 day, 14 days, and 28 days. Follow-up CT scans were performed on day 0, day 1, day 14, and day 28. Biochemical markers were collected before the procedure, 1 day after the procedure, and 14 days after the procedure. RESULTS: All ablation zones could be visualized on CT scan immediately after the procedure and on day 1 follow-up CT scan, and all animals survived until the designated endpoints. Histopathology revealed necrosis and edema on day 1 and fibrosis and glandular atrophy after 28 days. Blood vessels close to the ablation zone appeared normal. Laboratory analysis indicated mild to moderate amylasemia and lipasemia with normalization after 14 days. The ablation size on CT scan measured a mean (± SD) 146% ± 18 (day 0, P < .126) and 168% ± 18 (day 1, P < .026) of the simulation and on pathology measured 119% ± 10 (day 1, not significant) of the simulation. CONCLUSIONS: Results from simulations for planning IRE ablations, CT, and pathology may differ from each other. Ablation zones on CT and pathology appear larger than simulated, suggesting that clinically used treatment planning may underestimate the ablation size in the pancreas.

Evaluation of an endorectal electrode for performing focused irreversible electroporation ablations in the Swine rectum.

PURPOSE: To study the feasibility of a novel endorectal electrode for the creation of focal ablations of the rectal wall with the use of irreversible electroporation (IRE). MATERIALS AND METHODS: A monopolar electrode with a grounding pad (10 ablations in five pigs) and a bipolar electrode (two ablations in one pig) were evaluated in healthy swine rectum. A two-dimensional model of the electrode in the rectum was created and used to solve the Laplace equation to determine field strength. Simulation was used to identify treatment settings for superficial ablation (mucosal layers) or transmural ablation of rectal wall. Animals were euthanized within 4 hours after treatment. RESULTS: Treatment was successfully completed without treatment-related complications. Eleven of 12 lesions were successfully located and extracted for pathologic analysis. All lesions were characterized by necrotic cell death with mild inflammation and hyperemia, with a sharp demarcation between ablated and adjacent normal tissue. Depth of lesions corresponded with numeric simulation. Histologic analysis and measurements indicated that lesion creation with the superficial treatment setting resulted in ablation of mucosal and submucosal layers with superficial or no injury to the muscularis propria (9.97 mm ± 0.31 length, 3.3 mm ± 2.92 depth), and that lesion creation with the transmural treatment setting resulted in full-thickness ablation (12.43 mm ± 3.85 length, 4.97 mm ± 2.89 depth) of the rectal wall. CONCLUSIONS: An endorectal electrode can be used to deliver IRE and create limited focal ablations in the rectal wall. Treatment parameters can be determined through numeric modeling to control the depth of penetration of ablation.

Endoscopic spray cryotherapy for genitourinary malignancies: safety and efficacy in a porcine model.

OBJECTIVE: To examine the effects and safety of using endoscopic spray cryotherapy (ESC) on bladder, ureteral, and renal pelvis urothelium in a live porcine model. SUBJECTS AND METHODS: ESC treatments were systematically applied to urothelial sites in the bladder, ureter, and renal pelvis of eight female Yorkshire swine in a prospective trial. Freeze-thaw cycles ranged from 5 to 60 s/cycle for one to six cycles using a 7 French cryotherapy catheter. Tissue was evaluated histologically for treatment-related effects. Acute physiologic effects were evaluated with pulse oximetry, Doppler sonography, and postmortem findings. RESULTS: In bladder, treatment depth was inconsistent regardless of dose, demonstrating urothelial necrosis in one, muscularis propria depth necrosis in two, and full thickness necrosis in all remaining samples. In ureter, full thickness necrosis was seen in all samples, even with the shortest spray duration (5 s/cycle for six cycles or 30 s/cycle for one cycle). Treatment to the renal pelvis was complicated by adiabatic gas expansion of liquid nitrogen to its gaseous state, resulting in high intraluminal pressures requiring venting to avoid organ perforation, even at the lowest treatment settings. At a planned dose of 5 s/cycle for six cycles of the first renal pelvis animal, treatment was interrupted by sudden and unrecoverable cardiopulmonary failure after three cycles. Repeated studies replicated this event. Ultrasound and immediate necropsy confirmed the creation of a large gaseous embolism and reproducible cardiopulmonary effects. CONCLUSION: ESC in a porcine urothelial treatment model results in full-thickness tissue necrosis in bladder, ureter, and renal pelvis at a minimal treatment settings of 5 s/cycle for six cycles. Adiabatic gas expansion may result in fatal pyelovenous gas embolism and collateral organ injury, as seen in both animals receiving treatment to the renal pelvis in this study. These results raise safety concerns for use of ESC as a treatment modality in urothelial tissues with current device settings.

Neutrophil gelatinase-associated lipocalin (NGAL) levels in response to unilateral renal ischaemia in a novel pilot two-kidney porcine model.

OBJECTIVES: To test a novel porcine two-kidney model for evaluating the effect of controlled acute kidney injury (AKI) related to induced unilateral ischaemia on both renal units (RUs) To use neutrophil gelatinase-associated lipocalin (NGAL) and physiological serum and urinary markers to assess AKI and renal function. METHODS: Twelve female Yorkshire pigs had bilateral cutaneous ureterostomies placed laparoscopically with identical duration of pneumoperitoneum for all cases. An experimental group (n = 9) underwent induced unilateral renal ischaemia with left hilar clamping of timed duration (15, 30, 60 min) and a control group (n = 3) had no induced renal ischaemia. Urine was collected and analysed from each RU to assess creatinine and NGAL concentration preoperatively and at multiple postoperative time points. Serum was collected and analysed daily for creatinine and NGAL levels. Statistical comparisons were made using the rank-sum and sign-rank tests. RESULTS: Three pigs were excluded because of intra-operative and postoperative complications. In the RUs that experienced renal ischaemia (n = 7),the median urine volume was lower (P = 0.04) at 6, 12, 24 and 48 h and the median NGAL concentration was higher (P = 0.04) at 12 and 48 h compared with the RUs of control pigs that experienced no renal ischaemia (n = 2). When comparing the ischaemic (left) RU of the pigs in the experimental group with their contralateral non-ischaemic (right) RU, ischaemic RUs had a lower median cumulative urine volume at 6, 12, 24 and 48 h (P = 0.05) and a higher median NGAL concentration at 12, 24 and 48 h (P < 0.05). At 48 h, no significant increase was found in serum NGAL in pigs in the experimental group compared with controls (P = 0.2). Creatinine clearance (CC) was lower in ischaemic RUs compared with non-ischaemic RUs 1 day after surgery (P = 0.04) with decreasing CC as the duration of ischaemia increased. CONCLUSIONS: We have developed a promising novel small-scale pilot surgical model that allowed the evaluation of bilateral RU function separately during and after unilateral renal ischaemia. The induction of unilateral renal ischaemia corresponds with physiological changes in both the ischaemic and contralateral RU. AKI as measured by increases in NGAL and decreased renal function as measured by decreases in CC, are specific to the RU exposed to ischaemia.

Irreversible electroporation ablation of the liver can be detected with ultrasound B-mode and elastography.

BACKGROUND: Irreversible electroporation (IRE) is a novel ablation technique that induces permanent membrane permeability and cell death. We are interested in ultrasound B-mode and elastography to monitor IRE ablation in the liver. METHODS: Yorkshire pigs underwent IRE ablation of the liver and were imaged with ultrasound B-mode and elastography. Histologic evaluation of cell death by triphenyltetrazolium chloride and hematoxylin and eosin staining was performed. RESULTS: Elastography showed that liver ablated by IRE exhibited increased tissue stiffness with a peak strain ratio of 2.22. The IRE lesion had a discrete border without bubble artifact, and the lesion size significantly correlated with area of cell death on histology. IRE ablation was unaffected by presence of large blood vessels or bile ducts. CONCLUSION: IRE ablation led to increased tissue stiffness that was detectable by elastography and indicative of cell death. Elastography may complement B-mode ultrasonography to monitor IRE ablation of the liver.

Interstitial laser-induced thermotherapy of the lung: evaluation of the influence of ablation continuity on ablation size in a swine model.

The purpose of this study was to assess the relationship between size and the continuity of energy application in interstitial laser-induced thermotherapy. Percutaneous computed tomography-guided laser ablation (30 W, 600 nm diode) of the lung was performed in 7 Yorkshire pigs; a total of 42 ablation zones were created. Twenty ablations were performed using a continuous cycle of 2 min (protocol A) and 22 ablations were performed using 4 intermittent cycles with a duration of 1 min for each cycle interrupted by a 10-s stop between the cycles (protocol B). The lung was harvested immediately after euthanasia for gross pathology and histopathologic evaluation. Statistical analysis was performed using the Student t test and the Spearman correlation coefficient. Laser ablation resulted in complete necrosis of variable size of lung. The mean ablation zone dimensions (±SD) were 1.9 (±0.4) cm × 1.4 (±0.3) cm for protocol A and 2.2 (±0.5) cm × 1.4 (±0.4) cm for protocol B. The size of the necrosis is not significantly different when comparing a continuous 2-min ablation to a 4-cycle intermittent ablation for 1 min each cycle interrupted by a 10-s stop between the cycles (P = 0.98 and 0.53, respectively).

Irreversible electroporation adjacent to the rectum: evaluation of pathological effects in a pig model.

PURPOSE: To evaluate the effects of irreversible electroporation (IRE) on the rectum wall after IRE applied adjacent to the rectum. MATERIAL AND METHODS: CT-guided IRE adjacent to the rectum wall was performed in 11 pigs; a total of 44 lesions were created. In five pigs, ablations were performed without a water-filled endorectal coil (group A); in six pigs, ablation was performed with the coil to avoid displacement of the rectum wall (group B). The pigs were killed after 7-15 days and the rectums were harvested for pathological evaluation. RESULTS: There was no evidence of perforation on gross postmortem examination. Perirectal muscle lesions were observed in 18 of 20 ablations in group A and in 21 of 24 ablations in group B. Inflammation and fibrosis of the muscularis propria was observed in ten of 18 lesions in group A and in ten of 21 lesions in group B. In group A, findings were limited to the external layer of the muscularis propria except for one lesion; in group B, findings were transmural in all cases. Transmural necrosis with marked suppurative mucosal inflammation was observed in seven of 21 lesions in group B and in no lesion in group A. CONCLUSION: IRE-ablation adjacent to the rectum may be uneventful if the rectum wall is mobile and able to contract. IRE-ablation of the rectum may be harmful if the rectum wall is fixed adjacent to the IRE-probe.

The delayed effects of irreversible electroporation ablation on nerves.

OBJECTIVE: To evaluate the delayed effects of irreversible electroporation (IRE) ablation on nerves. METHODS: The study was approved by the institutional animal care and use committee. CT-guided IRE-ablation (electric field per distance, 1,500 V/cm; pulse length, 70 µs; number of pulses, 90) of 6 sciatic nerves was performed in 6 pigs that were euthanized 2 months after ablation. The sciatic nerves were harvested immediately after euthanasia for histopathological evaluation. Sections from selected specimens were stained with haematoxylin and eosin (H&E), Masson's trichrome (MT) method for collagen, and immunohistochemistry was performed for S100 and neurofilaments (markers for Schwann cells and axons, respectively). RESULTS: All nerves showed a preserved endoneural architecture and presence of numerous small calibre axons associated with Schwann cell hyperplasia, consistent with axonal regeneration. A fibrous scar was observed in the adjacent muscle tissue, confirming ablation at the site examined. CONCLUSION: After IRE-ablation of nerves, the preservation of the architecture of the endoneurium and the proliferation of Schwann cells may enable axonal regeneration as demonstrated after 2 months in this study.

Palatability and treatment efficacy of various ibuprofen formulations in C57BL/6 mice with ulcerative dermatitis.

Ibuprofen, a nonsteroidal antiinflammatory drug, is a nonselective inhibitor of cyclooxygenases 1 and 2 that commonly is used for its analgesic, antiinflammatory, and antipyretic properties. We compared the palatability and efficacy of medicated water containing ibuprofen from an oral pediatric suspension or liqui-gel capsules in C57BL/6 and genetically engineered mice of C57BL/6 background with ulcerative dermatitis. Mice (n = 14 or 15 per group) with ulcerated skin lesions of similar average size (capsule group, 6.71 mm(2); suspension group, 6.12 mm(2)) received ibuprofen in their drinking water at a concentration of 1 mg/mL. Water and food consumption, locomotor activity, grooming frequency, and reduction in pruritic behavior and lesion size were measured over a 9-d period. Compared with those treated with water containing the suspension, mice that received medicated water containing the liqui-gel formulation drank more (mean, 6.8 compared with 11.7 mL/d), consumed more food (4.02 compared with 2.73 g/d), and showed less pruritic behavior, greater healing (mean, 29.3% compare with 64.8%), and more locomotor activity over a 9-d period.

Flexible CO2 laser and submucosal gel injection for safe endoluminal resection in the intestines.

BACKGROUND: The CO(2) laser's unique wavelength of 10.6 µm has the advantage of being readily absorbed by water but historically limited it to line-of-sight procedures. Through recent technological advances, a flexible CO(2) laser fiber has been developed and holds promise for endoluminal surgery. We examined whether this laser, along with injection of a water-based gel in the submucosal space, will allow safe dissection of the intestines and enhance the potential of this tool for minimally invasive surgery. METHODS: Using an ex vivo model with porcine intestines, spot ablation was performed with the flexible CO(2) laser at different power settings until transmural perforation. Additionally, excisions of mucosal patches were performed by submucosal dissection with and without submucosal injection of a water-based gel. RESULTS: With spot ablation at 5 W, none of the specimens was perforated by 5 min, which was the maximum recorded time. The time to perforation was significantly shorter with increased laser power, and gel pretreatment protected the intestines against spot ablation, increasing the time to perforation from 6 to 37 s at 10 W and from 1 to 7 s at 15 W. During excision of mucosal patches, 56 and 83% of untreated intestines perforated at 5 and 10 W, respectively. Gel pretreatment prior to excision protected all intestines against perforation. These specimens were verified to be intact by inflation with air to over 100 mmHg. Furthermore, excision of the mucosal patch was complete in gel-pretreated specimens, whereas 22% of untreated specimens had residual islands of mucosa after excision. CONCLUSION: The flexible CO(2) laser holds promise as a precise dissection and cutting tool for endoluminal surgery of the intestines. Pretreatment with a submucosal injection of a water-based gel protects the intestines from perforation during ablation and mucosal dissection.