CT-based evaluation of tissue expansion in cryoablation of ex vivo kidney.

OBJECTIVES: To evaluate tissue expansion during cryoablation, the displacement of markers in ex vivo kidney tissue was determined using computed tomographic (CT) imaging. METHODS: CT-guided cryoablation was performed in nine porcine kidneys over a 10 min period. Markers and fiber optic temperature probes were positioned perpendicular to the cryoprobe shaft in an axial orientation. The temperature measurement was performed simultaneously with the acquisitions of the CT images in 5 s intervals. The distance change of the markers to the cryoprobe was determined in each CT image and equated to the measured temperature at the marker. RESULTS: The greatest increase in the distance between the markers and the cryoprobe was observed in the initial phase of cryoablation. The maximum displacement of the markers was determined to be 0.31±0.2 mm and 2.8±0.02 %, respectively. CONCLUSIONS: The mean expansion of ex vivo kidney tissue during cryoablation with a single cryoprobe is 0.31±0.2 mm. The results can be used for identification of basic parameters for optimization of therapy planning.

Distance between the descending aorta and the left inferior pulmonary vein as a determinant of biophysical parameters during paroxysmal atrial fibrillation cryoablation.

INTRODUCTION: The distance from the descending aorta (DA) to the posterior wall of the left atrium (LA) is variable. We aimed to determine whether the proximity between the DA and the left inferior pulmonary vein (LIPV) ostium has an impact on biophysical parameters and cryoballoon (CB) ablation efficacy during LIPV freezing. METHODS: Patients referred for CB-ablation of atrial fibrillation (AF) in two high-volume centers were included. Cryoablation data were collected prospectively for each patient. The anatomical relationships between the LIPV and the DA (distance LIPV ostium-DA, presence of an aortic imprint on the posterior aspect of the LIPV) were then retrospectively analysed on the LA computed tomography scans realized before AF ablation. RESULTS: A total of 350 patients were included (70% men, 59.7 ± 11.5 years). The decrease in the Ostium-DA distance was significantly correlated to the increase in the time-to-isolation (TTI) (r = -.31; p = .036), with less negative temperature (r = -.11; p = .045). Similarly, the presence of an aortic imprint on the LIPV was associated with a longer TTI (p < .001). The analysis of redo procedures data shows a trend toward the presence of shorter ostium-DA distances (15.3 ± 3.29 vs. 18.1 ± 4.99, p = .15) and more frequent aortic imprints (63.6% vs. 47.5%, p = .34) in patients with LIPV reconnection as opposed to patients without reconnection in the LIPV. CONCLUSION: Our findings indicated that the DA seems to have a "radiator" effect influencing LIPV cryoablation parameters during CB-ablation. Additional studies will be needed to elucidate whether this biophysical influence has a clinical impact in LIPVs reconnections.

Microglial cell-derived interleukin-6 influences behavior and inflammatory response in the brain following traumatic brain injury.

Traumatic brain injury (TBI) is a major health problem with high rates of mortality and morbidity worldwide. The response of the brain to TBI is orchestrated by a number of cytokines, including interleukin-6 (IL-6). IL-6 is a major cytokine in the central nervous system and it is produced by different cells, such as neurons, glial cells, and endothelial cells. Since glial cells are one of the most important sources and targets of IL-6, we have examined the role of microglia-derived IL-6 in normal conditions and following a model of TBI, cryolesion of the somatosensorial cortex. To this end, tamoxifen-inducible microglial IL-6-deficient (Il6ΔMic , using Cx3cr1 CreER model) mice and control (Il6lox/lox ) mice were used. In normal conditions, microglial IL-6 deficiency reduced deambulation and exploratory behavior and decreased anxiety in a sex-dependent manner. The transcriptome profile following cryolesion was dramatically altered 1 day post-lesion in Il6ΔMic compared with Il6lox/lox mice. However, the phenotype of Il6ΔMic mice was less compromised in the following days, suggesting that compensatory mechanisms are at play.

Evaluation of skeletal muscle regeneration in experimental model after malnutrition / Avaliação da regeneração do músculo esquelético em modelo experimental após desnutrição

Abstract The aim of this study was to analyze muscle regeneration after cryoinjury in the tibialis anterior muscle of young rats that were malnourished and then recovered. Forty Wistar rats were divided into a nourished group that received a normal protein diet (14% casein) for 90 days and a malnourished and recovered rats group (MR) that was submitted to 45 days of malnutrition with a hypoproteic diet (6% casein) followed by 45 days of a normal protein diet (14% casein). After the recovery period, all of the animals underwent cryoinjury in the right tibialis anterior muscle and euthanasia after 7, 14 and 21 days. The amount of connective tissue and the inflammation area was higher in the malnutrition recovered injury MR group (MRI) at 14 days post-injury (p < 0.05). Additionally, the cross-sectional area (CSA) of the regenerated fibers was decreased in the MRI (p < 0.05). The MyoD and myogenin protein levels were higher in the nourished injury group. Similar levels of TGF-β1 were found between groups. The proposed malnutrition protocol was effective in showing delayed changes in the regeneration process of the tibialis anterior muscle of young rats. Furthermore, we observed a delay in muscle repair even after nutritional recovery.

Resumo O objetivo do presente estudo foi analisar a regeneração muscular após criolesão no músculo tibial anterior de ratos jovens desnutridos e recuperados. Foram utilizados 40 ratos da linhagem Wistar, divididos em 2 grupos: ratos nutridos receberam dieta normoproteica (14% de caseína) por 90 dias; e ratos desnutridos e recuperado submetidos a duas fases nutricionais pós-desmame, correspondendo a 45 dias de desnutrição com dieta hipoproteica (6% caseína), seguida por 45 dias de dieta normoproteica (14% caseína). Ao completar a fase de recuperação, todos os animais foram submetidos à criolesão no músculo tibial anterior direito e a eutanasia ocorreu 7, 14 e 21 dias após a lesão. A quantidade de tecido conjuntivo e a área de inflamação 14 dias pós-lesão foi maior no grupo desnutrido, recuperado e lesado (MRI – malnourished, recovered and injured group) (p < 0,05). A área de secção transversa (AST) das fibras regeneradas do grupo MRI foi menor (p < 0,05). O conteúdo das proteínas MyoD e Miogenina foi maior no grupo nutridos e lesados. A citocina TGF-β1 não apresentou diferença entre os grupos. O protocolo proposto foi eficaz para demonstrar alterações no processo de regeneração do músculo tibial anterior de ratos jovens, atrasando o reparo muscular mesmo após a recuperação nutricional.

Zebrafish fin regeneration after cryoinjury-induced tissue damage.

Although fin regeneration following an amputation procedure has been well characterized, little is known about the impact of prolonged tissue damage on the execution of the regenerative programme in the zebrafish appendages. To induce histolytic processes in the caudal fin, we developed a new cryolesion model that combines the detrimental effects of freezing/thawing and ischemia. In contrast to the common transection model, the damaged part of the fin was spontaneously shed within two days after cryoinjury. The remaining stump contained a distorted margin with a mixture of dead material and healthy cells that concomitantly induced two opposing processes of tissue debris degradation and cellular proliferation, respectively. Between two and seven days after cryoinjury, this reparative/proliferative phase was morphologically featured by displaced fragments of broken bones. A blastemal marker msxB was induced in the intact mesenchyme below the damaged stump margin. Live imaging of epithelial and osteoblastic transgenic reporter lines revealed that the tissue-specific regenerative programmes were initiated after the clearance of damaged material. Despite histolytic perturbation during the first week after cryoinjury, the fin regeneration resumed and was completed without further alteration in comparison to the simple amputation model. This model reveals the powerful ability of the zebrafish to restore the original appendage architecture after the extended histolysis of the stump.

Neural tissue regeneration in experimental brain injury model with channeled scaffolds of acrylate copolymers.

The objective of the present study was to evaluate the biocompatibility and cell hosting ability of a copolymer scaffold based on ethyl acrylate (EA) and hydroxyl ethyl acrylate (HEA) in vivo after an experimental brain injury. Wistar rats were subjected to cryogenic traumatic brain injury. We evaluated the tissue response to the implanted materials after 8 weeks. The materials were implanted devoid of cells; they provoked a minimal scar response by the host tissue and permitted the invasion of neurons and glia inside them. We also found new blood vessels surrounding and inside the implant. Thus, the copolymer scaffold proves to offer a suitable environment producing a cellular network potentially useful in brain repair after brain injury.

Regenerated soleus muscle shows reduced creatine kinase efflux after contractile activity in vitro.

Regenerated skeletal muscles show less muscle damage after strenuous muscle exercise. The aim of the studies was to investigate if the regeneration is associated with reduced muscle creatine kinase (CK) efflux immediately after the exercise. Cryolesion was applied to the soleus muscle of 3-month-old C57BL/6J male mice. Then total CK efflux was assessed in vitro in the regenerated muscles without exercise or after 100 eccentric contractions. The same measurements were performed in the control muscles, which were not exposed to cryolesion. Regenerated muscles generated weaker (P < 0.05) twitches, but stronger (P < 0.05) 150-Hz and 300-Hz tetani with prolonged (P < 0.01) contraction times compared with the control muscles. There was no difference between regenerated and control muscles in the total CK efflux without exercise, but only control muscles showed an increase (P < 0.001) in the CK efflux after the exercise. Our results suggest that muscle regeneration is associated with modulation of contractile properties and improvement in muscle resistance to damage after eccentric exercise.

Morphological aspects and Cox-2 expression after exposure to 780-nm laser therapy in injured skeletal muscle: an in vivo study

Background: The effectiveness of low-level laser therapy in muscle regeneration is still not well known. Objective: To investigate the effects of laser irradiation during muscle healing. Method: For this purpose, 63 rats were distributed to 3 groups: non-irradiated control group (CG); group irradiated at 10 J/cm² (G10); and group irradiated at 50 J/cm² (G50). Each group was divided into 3 different subgroups (n=7), and on days 7, 14 and 21 post-injury the rats were sacrificed. Results: Seven days post-surgery, the CG showed destroyed zones and extensive myofibrillar degeneration. For both treated groups, the necrosis area was smaller compared to the CG. On day 14 post-injury, treated groups demonstrated better tissue organization, with newly formed muscle fibers compared to the CG. On the 21st day, the irradiated groups showed similar patterns of tissue repair, with improved muscle structure at the site of the injury, resembling uninjured muscle tissue organization. Regarding collagen deposition, the G10 showed an increase in collagen synthesis. In the last period evaluated, both treated groups showed statistically higher values in comparison with the CG. Furthermore, laser irradiation at 10 J/cm2 produced a down-regulation of cyclooxygenase 2 (Cox-2) immunoexpression on day 7 post-injury. Moreover, Cox-2 immunoexpression was decreased in both treated groups on day 14. Conclusions: Laser therapy at both fluencies stimulated muscle repair through the formation of new muscle fiber, increase in collagen synthesis, and down-regulation of Cox-2 expression. .

Effects of 660 nm low-level laser therapy on muscle healing process after cryolesion.

The aim of this study was to evaluate the effects of 660 nm low-level laser therapy (LLLT) on muscle regeneration after cryolesion in rat tibialis anterior muscle. Sixty-three Wistar rats were divided into a control group, 10 J/cm(2) laser-treated group, and 50 J/cm(2) laser-treated group. Each group formed three subgroups (n = 7 per group), and the animals were sacrificed 7, 14, or 21 d after lesion. Histopathological findings revealed a lower inflammatory process in the laser-treated groups after 7 d. After 14 d, irradiated animals at both fluences showed higher granulation tissue, new muscle fibers, and organized muscle structure. After 21 d, full tissue repair was observed in all groups. Moreover, irradiated animals at both fluences showed smaller necrosis area in the first experimental period evaluated. MyoD immunoexpression was observed in both treated groups 7 d postinjury. Myogenin immunoexpression was detected after 7 and 14 d. The higher fluence increased the number of blood vessels after 14 and 21 d. These results suggest that LLLT, at both fluences, positively affects injured skeletal muscle in rats, accelerating the muscle-regeneration process.

Effects of low-intensity pulsed ultrasound on injured skeletal muscle

BACKGROUND: Low-intensity pulsed ultrasound (LIPUS) has been shown to stimulate tissue metabolism and accelerate muscle healing. However, the optimal parameters in the use of LIPUS are still not clear. OBJECTIVE: The aim of this study was to analyze the effects of LIPUS on muscle healing in rats subjected to a cryolesion. METHOD: Twenty rats were divided into the following groups: an injured control group (CG) and an injured treated group (TG). Both groups were divided into 2 sub-groups (n=5 each) that were sacrificed 7 and 13 days post-surgery. Treatments were started 24 hours after the surgical procedure and consisted of 3 or 6 sessions. After euthanasia, the muscles were submitted to standard histological procedures. RESULTS: Qualitative analyses were based on morphological assessments of the muscle. The histopathological analysis on day 7 revealed that the muscles in the CG and the TG presented an intense inflammatory infiltrate, a large necrotic area and a disorganized tissue structure. After 13 days, both the CG and the TG had granulation tissue and newly formed fibers. The TG presented a more organized tissue structure. The quantitative analysis of collagen indicated similar findings among the groups, although the qualitative analysis revealed a better organization of collagen fibers in the TG at 13 days. The immunohistochemical analysis indicated that, at both time points, the expression of cyclooxygenase-2 was upregulated in the TG compared to the CG. CONCLUSIONS: LIPUS used as a treatment for muscle injury induced a more organized tissue structure at the site of the injury and stimulated the expression of COX-2 and the formation of new muscle fibers. .