Impact of prophylactic corticosteroids on systemic inflammation after extensive atrial ablation in pigs.

AIMS: Prophylactic corticosteroids have been reported to attenuate the increase in C-reactive protein (CRP) and the incidence of atrial fibrillation (AF) both after heart surgery and AF ablation. We tested the impact of a single prophylactic corticosteroid dose on ultrasensitive CRP 24 h and 14 days after extensive linear atrial ablation (8 mm or 3.5 irrigated tip) guided by electroanatomical mapping (NavX) in pigs with normal hearts. METHODS AND RESULTS: Pigs (n = 19; 35 kg) were divided into three groups: corticoid (n = 7), atrial ablation with administration of 500 mg methylprednisolone intravenous at anaesthetic induction; control (n = 7), atrial ablation only; and sham (n = 5), surgical procedure without ablation. Troponin and CRP were measured before, 24 h and 14 days after the procedure. After sacrifice, lesions were analysed macroscopically and histologically. Linear lesions were created in the right (n = 23) and left (n = 21) atrium of 14 animals, with no difference between groups. In all groups there was elevation of troponin and CRP 24 h after ablation, with a return to baseline values after 14 days. However, CRP levels of the control, corticoid, and sham groups were similar at all three time points analysed (baseline P = 0.52, 24 h P = 0.21, 14 days P = 0.66). Histological analysis did not show any difference between corticoid and control groups. CONCLUSION: In this model, extensive biatrial RF ablation, per se, does not promote systemic inflammation. The use of a prophylactic single corticoid dose before ablation did not prevent systemic inflammation or alter the healing of the lesions.

Radiofrequency ablation does not induce apoptosis in the rat myocardium.

BACKGROUND: The mechanisms implicated in the genesis of delayed radiofrequency (RF) effects remain unclear, but may be related to extension of the lesion beyond the region of coagulative necrosis. The role of apoptosis in this process has not been previously reported. We assessed whether RF promotes apoptosis in the region surrounding acute ablation lesions in a rat model. METHODS: Wistar rats (n=30; weight 300 g) were anesthesized, the chest was opened, and the heart was exposed. A modified unipolar RF ablation (custom catheter 4.5-mm-tip diameter, 12 Watts, 10 seconds) was undertaken on the left ventricular anterolateral epicardial surface and the chest was closed. After 2 hours, animals were killed for histological (hematoxylin and eosin, TdT-mediated dUTP Nick End-Labeling [TUNEL] assay) and immunohistochemical (anti-BAD and anti-caspase 3 antibodies) analysis (n=18). Additional animals (n=12) were sacrificed at 2 (n=3), 24 (n=3), 48 (n=3), and 72 hours (n=3) after ablation exclusively for anti-BAD Western Blotting analysis. RESULTS: Lesions were characterized by well-defined regions of coagulative necrosis. In 18/18 (100%) animals, TUNEL assay revealed positive luminescent reaction cells in the region surrounding the lesion, extending up to 2 mm from the border zone. However, microscopic evaluation of the nuclei and immunohistochemical and anti-BAD Western Blotting analysis were negative in all (100%) rats. Thus, positive TUNEL reaction in the periphery of the ablation lesion likely reflects nonspecific DNA damage. CONCLUSION: RF ablation does not promote apoptosis in the periphery of the myocardial lesion. This finding may have implications for the elucidation of late lesion extension following RF ablation.

Impact of corticosteroids on late growth of radiofrequency lesions in infant pigs: histopathological and electroanatomical findings.

AIMS: Corticosteroids attenuate late growth of radiofrequency (RF) lesions in the thigh muscle of infant rats. We sought to assess the impact of these drugs on the late growth of RF lesions in immature swine myocardium and to determine the electroanatomical mapping (EAM) characteristics of these lesions. METHODS AND RESULTS: Radiofrequency (60°C; 60 s) lesions were created in the right atrium (n = 2) and ventricle (n = 2) of 14 piglets (age 65 days; weight 5 kg) and 3 adults. Piglets were divided into: controls (n = 7) and treated (n = 7), receiving hydrocortisone (10 mg/kg iv after RF) and prednisone (1 mg/kg/day) for 29 days. After 8 months, animals were sacrificed for histological analysis. In four piglets, endocardial and epicardial voltage EAM were performed. In infant groups, the dimensions of atrial (11 ± 5 vs. 13 ± 7 mm) and ventricular (12 ± 3 vs. 11 ± 3 mm) lesions were similar. In adults, atrial (6 ± 1 mm) and ventricular (6 ± 1 mm) lesions were smaller. In controls, ventricular lesions depicted dense fibrosis and multiple strands of fibrous tissue extending from the lesion into normal muscle. Treated piglets revealed scars exhibiting less dense fibrosis with predominance of fibroadipose tissue and less collagen proliferation. Large atrial and ventricular low-voltage areas corresponding to the macroscopic lesions were identified in all animals. CONCLUSION: Radiofrequency lesions in infant pigs reveal late growth and invasion of normal muscle by intense collagen proliferation. Corticosteroids do not prevent late enlargement of the lesions but modulate the fibrotic proliferation. The expressive growth of the lesion may generate low-voltage areas detectable by EAM.

Effects of corticosteroid therapy on the long-term outcome of radiofrequency lesions in the Swine caval veins.

BACKGROUND: We explored the angiographic and pathological effects of corticosteroids on the long-term outcome of radiofrequency (RF) ablation lesions in the swine caval veins. METHODS: Under fluoroscopy guidance, a single linear RF lesion (4-mm tip, 60 degrees C, 180 seconds) was created in each vena cava (from +/-2 cm into the vein to the venoatrial junction) of 20 anesthetized minipigs (35+/- 2 kg). Three groups were studied: acute (n = 4), killed 1 hour after RF; control (n = 8), sacrificed 83+/- 1 days after RF; and pigs (n = 8) receiving hydrocortisone (400 mg i.v. after RF) and prednisone (25 mg po for 30 days), killed 83+/- 1 days post-RF. Angiography was performed before, immediately after ablation, and at follow-up. Then, animals were sacrificed for histological analysis. RESULTS: Mild (<40%) or moderate (41-70%) acute luminal narrowing occurred in 19/20 (95%) inferior veins and in 13/20 (65%) superior veins. Severe (>70%) stenosis and occlusions were not noted. At follow-up, in both chronic groups, mean vessel diameters returned to baseline and progression of luminal narrowing did not occur in any vein. Of note, superior and inferior vena cava angiographic diameter for control and treated pigs did not differ. The same was observed for the cross-sectional luminal area. Acute lesions displayed transmural coagulative necrosis whereas chronic lesions revealed marked fibrosis. Histological findings were similar in controls and treated pigs. CONCLUSION: In this model, mild and moderate stenosis, occurring immediately after ablation, seems to resolve over time. Corticosteroids do not affect the long-term outcome of such RF lesions in the caval veins.

Effect of atenolol pre-treatment in heart damage in a model of intestinal ischemia-reperfusion.

PURPOSE: To investigate the effects of atenolol in inflammatory mediator and oxidative stress in a myocardial injury by intestinal ischemia/reperfusion in rat model. METHODS: Adult Wistar male rats were randomly (n=8), anesthetized and divided in: Sham: submitted to operation only; group SS+IR: intravenous saline infusion following superior mesenteric artery occlusion during 60 minutes (ischemia) and open for 120 minutes (reperfusion); group AT+IR: intravenous atenolol infusion (2 mg/kg) following superior mesenteric artery occlusion during 60 minutes (ischemia) and open for 120 minutes (reperfusion); and group AT+I+AT+R: intravenous atenolol infusion following superior mesenteric artery occlusion during 60 minutes (ischemia) and in the time 45 minutes other atenolol doses were administrated and the artery was open for 120 minutes (reperfusion), all animals were submitted to muscular relaxation for mechanical ventilation. In the end of experiment the animals were euthanized and the hearts tissue were morphology analyzed by histology and malondialdehyde by ELISA, and the plasma were analyzed for tumor necrosis factor-alpha by ELISA. RESULTS: The group SS+IR demonstrated the higher malondialdehyde levels when compared with the atenolol treated-groups (p=0.001) in the heart tissue. The tumor necrosis factor-alpha level in plasma decrease in the treated groups when compared with SS+IR group (p=0.001). Histology analyses demonstrate pyknosis, edema, cellular vacuolization, presence of inflammatory infiltrate and band contraction in the heart tissue of the rats. CONCLUSION: Atenolol significantly reduce the degree of cardiac damage after intestinal ischemia-reperfusion.

Are the virtual lines created with the EnSite electroanatomical mapping system really continuous?

BACKGROUND: EnSiteNavx electroanatomic mapping system is widely used in radiofrequency (RF) atrial fibrillation ablation, helping the creation of linear lesions. However, the correspondence of the virtual line created by EnSite with the pathological lesion has not yet been evaluated. OBJECTIVE: to assess the continuousness of Ensite-guided virtual lines in a swine model. METHODS: we performed RF ablation linear lesions (8mm and irrigated catheters tips) in both atria of 14 pigs (35 Kg) guided by the EnSite. The animals were sacrificed 14 days post-ablation for macroscopic and histological analysis. RESULTS: a total of 23 lines in the right atrium and 21 lines in the left atrium were created in 14 animals. The medium power, impedance and temperature applications were 56 W, 54 ºC and 231 Ω for the 8mm tip, and 39 W, 37 ºC, 194 Ω for the irrigated tip catheter, respectively. All (100%) lines were identified on the epicardial and endocardial surfaces, denoting transmurality. At macroscopic examination, lesions were extensive and pale, continuous, with 3.61 cm long and 0.71 cm deep. The transmurality of the lesions was confirmed by microscopy. There was a correlation in the location of the lines at the virtual map and the anatomical lesions in 21 of 23 (91.3%) of the right atrium and 19/21 (90.4%) of the left atrium. CONCLUSION: In this model, the lines created in the virtual map by EnSiteNavX system correspond to continuous transmural linear lesions in anatomical specimen, suggesting that this method is suitable for linear ablation of atrial fibrillation.

Thermotolerance does not reduce the size or remodeling of radiofrequency lesions in the rat myocardium.

PURPOSE: Late lesion extension may be involved in the genesis of delayed radiofrequency (RF) effects. Because RF lesion is thermally mediated, we hypothesized that induction of heat shock response (thermotolerance) would modulate lesion healing. We evaluated the effects of thermotolerance on the dimensions and remodeling of RF lesions in a rat model of heart failure. METHODS: Wistar rats (weight 300 g) subjected to heat stress (n = 22, internal temperature of 42 °C for 10 min) were compared to controls (n = 22, internal temperature of 37 °C for 10 min). After 48 h (peak of HSP70 myocardial concentration), a modified unipolar RF lesion (customized catheter, tip 4.5 mm in diameter; 12 W; 10 s) was created on the left ventricular free wall. Animals were sacrificed 2 h (n = 10 per group) and 4 weeks (n = 12 per group) after ablation for lesion analysis. An echocardiogram was obtained at 4 weeks. RESULTS: There was no difference between groups regarding the size of acute (controls 27 ± 2 vs. treated 27 ± 3 mm(2)) and chronic lesions (controls 17 ± 1 vs. treated 19 ± 1 mm(2)). Histology of lesions did not differ between groups. The echocardiogram revealed dilation of the cavities and moderate systolic dysfunction without difference between groups. Acute lesion dimensions were similar between control and treated animals over time (ablation undertaken 3, 12, 24, 48, and 72 h after hyperthermia) and also using a conventional ablation catheter (50 °C; 15 W; 10 s). CONCLUSION: Thermotolerance does not reduce the size or remodeling of RF lesions in the rat myocardium.

Effect of atenolol pre-treatment in heart damage in a model of intestinal ischemia-reperfusion

Abstract Purpose: To investigate the effects of atenolol in inflammatory mediator and oxidative stress in a myocardial injury by intestinal ischemia/reperfusion in rat model. Methods: Adult Wistar male rats were randomly (n=8), anesthetized and divided in: Sham: submitted to operation only; group SS+IR: intravenous saline infusion following superior mesenteric artery occlusion during 60 minutes (ischemia) and open for 120 minutes (reperfusion); group AT+IR: intravenous atenolol infusion (2 mg/kg) following superior mesenteric artery occlusion during 60 minutes (ischemia) and open for 120 minutes (reperfusion); and group AT+I+AT+R: intravenous atenolol infusion following superior mesenteric artery occlusion during 60 minutes (ischemia) and in the time 45 minutes other atenolol doses were administrated and the artery was open for 120 minutes (reperfusion), all animals were submitted to muscular relaxation for mechanical ventilation. In the end of experiment the animals were euthanized and the hearts tissue were morphology analyzed by histology and malondialdehyde by ELISA, and the plasma were analyzed for tumor necrosis factor-alpha by ELISA. Results: The group SS+IR demonstrated the higher malondialdehyde levels when compared with the atenolol treated-groups (p=0.001) in the heart tissue. The tumor necrosis factor-alpha level in plasma decrease in the treated groups when compared with SS+IR group (p=0.001). Histology analyses demonstrate pyknosis, edema, cellular vacuolization, presence of inflammatory infiltrate and band contraction in the heart tissue of the rats. Conclusion: Atenolol significantly reduce the degree of cardiac damage after intestinal ischemia-reperfusion.

Effects of the corticosteroids in the lesions by radiofrequency on rats' thigh in different age groups.

BACKGROUND: The corticosteroids limit the late growth of the lesions by radiofrequency (RF) on the rats' infants' thighs, but the effects on the pubescent and adult rats are unknown. OBJECTIVE: Evaluate the effects of the corticosteroids in the healing of the lesions by RF on the rats' thighs muscles in different age groups. METHODS: Ablation was performed on the thigh muscle of 30 rats (1 lesion per animal): infants (30 days old, weight 73 g, n = 10), pubescents (60 days old, weight 230 g, n = 10) and 10 adults (90 days old, 310 g, n = 10), subdivided in control and treated groups, that received Hydrocortisone (10 mg/kg IM post-RF) and Betametasone (3.5 mg/kg IM, twice a week, for 29 days). The rats were sacrificed 60 days after the ablation for hystopathological and planimetric analysis with specified software (ImageJ). RESULTS: In the infant, pubescent and adult groups, the weight gain in the follow up did not differ between the control and the treated ones. In the control group, the lesions of the infants and pubescents were superior (p = 0.01) to the adults'. The treatment reduced the size of the lesion in the infants (5.58+0.61 mm² vs 4.02+0.23 mm(2); p < 0.01) and pubescents (5.20+0.47 mm(2) vs 4.16+0.48 mm(2); p < 0.01), but not in the adults (4.44+0.50 mm(2) vs 4.79+0.53 mm(2), p = NS). Infant and pubescent treated groups presented lower collagen deposition and less fibrotic bands invading the healthy tissue from the central fibrosis area, and forming lesions with remarkably more reduced dimensions than their controls. There were no differences in the adult groups. CONCLUSION: The corticosteroids seem to reduce the late growth of lesion, in addition to attenuate the fibrotic proliferation in the infant and pubescent rats.

As linhas virtuais criadas com o sistema de mapeamento eletroanatômico ensite são de fato contínuas? / Are the virtual lines created with the ensite electroanatomical mapping system really continuous?

FUNDAMENTO: O sistema de mapeamento eletro anatômico Ensite Navx é muito usado na ablação por radiofrequência (RF) da fibrilação atrial, ajudando na confecção de lesões lineares. Contudo, a correspondência da linha virtual criada pelo Ensite com a lesão patológica ainda não foi avaliada. OBJETIVO: Avaliar a continuidade da linha virtual criada pelo Ensite em modelo suíno. MÉTODOS: Realizamos ablação linear por RF (cateter de 8 mm e irrigado) em ambos os átrios de 14 suínos (35 kg) guiada pelo EnSite. Os animais foram sacrificados 14 dias pós-ablação para análise macroscópica e histológica. RESULTADOS: Foram confeccionadas 23 lesões lineares em átrio direito e 21 em átrio esquerdo dos 14 animais. A potência, temperatura e impedância médias das aplicações foram de 56 W, 54 ºC e 231 Ω para o cateter de 8mm, e de 39 W, 37 ºC e 194 Ω para o cateter irrigado. Todas (100%) as linhas foram identificadas nas faces epicárdica e endocárdica, denotando transmuralidade. À macroscopia, as lesões eram extensas e pálidas, com 3,61 cm de comprimento e 0,71 cm de profundidade e contínuas. A transmuralidade das lesões foi confirmada pela microscopia. Houve correspondência na localização das linhas do mapa virtual com as da peça anatômica em 21 das 23 (91,3%) das linhas do átrio direito e 19/21 (90,4%) do átrio esquerdo. CONCLUSÃO: Nesse modelo, as linhas criadas no mapa virtual pelo sistema EnSiteNavX se correlacionam a lesões lineares transmurais contínuas na peça anatômica, sugerindo que esse método é adequado para a ablação linear da fibrilação atrial.

BACKGROUND: EnSiteNavx electroanatomic mapping system is widely used in radiofrequency (RF) atrial fibrillation ablation, helping the creation of linear lesions. However, the correspondence of the virtual line created by EnSite with the pathological lesion has not yet been evaluated. OBJECTIVE: to assess the continuousness of Ensite-guided virtual lines in a swine model. METHODS: we performed RF ablation linear lesions (8mm and irrigated catheters tips) in both atria of 14 pigs (35Kg) guided by the EnSite. The animals were sacrificed 14 days post-ablation for macroscopic and histological analysis. RESULTS: a total of 23 lines in the right atrium and 21 lines in the left atrium were created in 14 animals. The medium power, impedance and temperature applications were 56 W, 54 ºC and 231 Ω for the 8mm tip, and 39W, 37ºC, 194 Ω for the irrigated tip catheter, respectively. All (100%) lines were identified on the epicardial and endocardial surfaces, denoting transmurality. At macroscopic examination, lesions were extensive and pale, continuous, with 3.61 cm long and 0.71 cm deep. The transmurality of the lesions was confirmed by microscopy. There was a correlation in the location of the lines at the virtual map and the anatomical lesions in 21 of 23 (91.3%) of the right atrium and 19/21 (90.4%) of the left atrium. CONCLUSION: In this model, the lines created in the virtual map by EnSiteNavX system correspond to continuous transmural linear lesions in anatomical specimen, suggesting that this method is suitable for linear ablation of atrial fibrillation.

Acute and chronic effects of extensive radiofrequency lesions in the canine caval veins: implications for ablation of atrial arrhythmias.

BACKGROUND: Although radiofrequency (RF) ablation within the caval veins has been increasingly used to treat a variety of atrial tachyarrhythmias, the consequences of RF ablation in the caval veins are unknown. We explored the acute and chronic angiographic and pathological effects of extensive RF ablation in the caval veins. METHODS: Under fluoroscopy guidance, conventional (4 mm tip, 60 degrees C, 60 seconds) RF applications (n = 6-7) were delivered in each vena cava (from +/-2 cm into the vein to the veno-atrial junction) of 15 dogs (10 +/- 3 kg). Animals were killed 1 hour and 5 weeks after ablation for histological analysis. Angiography was performed before ablation (acute dogs only) and at sacrifice to assess the degree of vascular stenosis. RESULTS: In acute dogs (n = 5), luminal narrowing was noted in 10/10 (100%) targeted veins (mild in two; moderate in three and severe in five, including two total occlusions). In the six chronic animals that completed the protocol (four died during follow-up), stenosis was also observed in 12/12 (100%) ablated veins (mild in six; moderate in four and severe in two). Of these, one superior vena cava was suboccluded with development of extensive collateral circulation. Histologically, acute lesions displayed typical transmural coagulative necrosis, whereas chronic lesions revealed intimal proliferation, necrotic muscle replaced with collagen, endovascular contraction, and disruption and thickening of the internal elastic lamina. CONCLUSION: In this model, extensive RF ablation in the caval veins may result in significant vascular stenosis. These findings may have implications for catheter ablation of arrhythmias originating within the caval veins.

Efeitos dos corticoesteroides nas lesões por radiofrequência na coxa de ratos em diferentes faixas etárias / Effects of the corticosteroids in the lesions by radiofrequency on rats' thigh in different age groups

FUNDAMENTO: Os corticosteroides limitam o crescimento tardio das lesões por radiofrequência (RF) no músculo da coxa de ratos filhotes, mas seus efeitos em ratos púberes e adultos são desconhecidos. OBJETIVO: Avaliar os efeitos dos corticoesteroides na cicatrização das lesões por RF no músculo da coxa de ratos em diferentes faixas etárias. MÉTODOS: Realizou-se ablação no músculo da coxa de 30 ratos (1 lesão por animal): infantis (30 dias de idade, peso 73 g, n = 10), púberes (60 dias de idade, peso 230 g, n = 10) e 10 adultos (90 dias de idade, 310 g, n = 10), subdivididos em controles e tratados, que receberam Hidrocortisona (10 mg/kg IM pós-RF) e Betametasona (3,5 mg/kg IM, duas vezes por semana, por 29 dias). Os ratos foram sacrificados 60 dias após a ablação para análise histopatológica e planimétrica com software específico (ImageJ®). RESULTADOS: Nos infantis, púberes e adultos o ganho de peso no seguimento não diferiu entre controles e tratados. Nos controles, as lesões dos infantis e púberes eram maiores (p = 0,01) que as dos adultos. O tratamento reduziu o tamanho da lesão nos infantis (5,58+0,61 mm² vs 4,02+0,23 mm²; p < 0,01) e nos púberes (5,20+0,47 mm² vs 4,16+0,48 mm²; p < 0,01), mas não nos adultos (4,44+0,50 mm² vs 4,79+0,53 mm², p = NS). Tratados Infantil e Púbere apresentaram menor deposição de colágeno e menos traves fibróticas invadindo o tecido saudável a partir da área de fibrose central, e compondo lesão de dimensões notadamente mais reduzidas que seus controles. Não houve diferenças nos grupos adultos. CONCLUSÃO: Os corticoesteroides parecem reduzir o crescimento tardio da lesão, além de atenuar a proliferação fibrótica nos ratos infantis e púberes.

BACKGROUND: The corticosteroids limit the late growth of the lesions by radiofrequency (RF) on the rats' infants' thighs, but the effects on the pubescent and adult rats are unknown. OBJECTIVE: Evaluate the effects of the corticosteroids in the healing of the lesions by RF on the rats' thighs muscles in different age groups. METHODS: Ablation was performed on the thigh muscle of 30 rats (1 lesion per animal): infants (30 days old, weight 73 g, n = 10), pubescents (60 days old, weight 230 g, n = 10) and 10 adults (90 days old, 310 g, n = 10), subdivided in control and treated groups, that received Hydrocortisone (10 mg/kg IM post-RF) and Betametasone (3.5 mg/kg IM, twice a week, for 29 days). The rats were sacrificed 60 days after the ablation for hystopathological and planimetric analysis with specified software (ImageJ®). RESULTS: In the infant, pubescent and adult groups, the weight gain in the follow up did not differ between the control and the treated ones. In the control group, the lesions of the infants and pubescents were superior (p = 0.01) to the adults'. The treatment reduced the size of the lesion in the infants (5.58+0.61 mm² vs 4.02+0.23 mm²; p < 0.01) and pubescents (5.20+0.47 mm² vs 4.16+0.48 mm²; p < 0.01), but not in the adults (4.44+0.50 mm² vs 4.79+0.53 mm², p = NS). Infant and pubescent treated groups presented lower collagen deposition and less fibrotic bands invading the healthy tissue from the central fibrosis area, and forming lesions with remarkably more reduced dimensions than their controls. There were no differences in the adult groups. CONCLUSION: The corticosteroids seem to reduce the late growth of lesion, in addition to attenuate the fibrotic proliferation in the infant and pubescent rats.