RESUMO
BACKGROUND AND OBJECTIVE: It is unknown if pro- and anti-inflammatory mediators in acute lung inflammation induced by intestinal ischemia and reperfusion (i-I/R) can be modulated by low-level laser therapy (LLLT). STUDY DESIGN/MATERIAL AND METHODS: A controlled ex vivo study was developed in which rats were irradiated (660 nm, 30 mW, 0.08 cm² of spot size) on the skin over the right upper bronchus 1 hour post-mesenteric artery occlusion and euthanized 4 hours later. For pretreatment with anti-tumor necrosis factor (TNF) or IL-10 antibodies, the rats received either one of the agents 15 minutes before the beginning of reperfusion. METHODS: Lung edema was measured by the Evans blue extravasation and pulmonary neutrophils influx was determined by myeloperoxidase (MPO) activity. Both TNF and IL-10 expression and protein in lung were evaluated by RT-PCR and ELISA, respectively. RESULTS: LLLT reduced the edema (80.1 ± 41.8 µg g⻹ dry weight), neutrophils influx (0.83 ± 0.02 × 106 cells ml⻹), MPO activity (2.91 ± 0.60), and TNF (153.0 ± 21.0 pg mg⻹ tissue) in lung when compared with respective control groups. Surprisingly, the LLLT increased the IL-10 (0.65 ± 0.13) in lung from animals subjected to i-I/R. Moreover, LLLT (0.32 ± 0.07 pg ml⻹) reduced the TNF-α level in RPAECs when compared with i-I/R group. The presence of anti-TNF or IL-10 antibodies did not alter the LLLT effect on IL-10 (465.1 ± 21.0 pg mg⻹ tissue) or TNF (223.5 ± 21.0 pg mg⻹ tissue) in lung from animals submitted to i-I/R. CONCLUSION: The results indicate that the LLLT attenuates the i-I/R-induced acute lung inflammation which favor the IL-10 production and reduce TNF generation.
Assuntos
Interleucina-10/biossíntese , Intestinos/irrigação sanguínea , Isquemia/complicações , Terapia com Luz de Baixa Intensidade , Pneumonia/radioterapia , Reperfusão , Fator de Necrose Tumoral alfa/biossíntese , Doença Aguda , Animais , Edema , Lasers Semicondutores/uso terapêutico , Pulmão/metabolismo , Pulmão/patologia , Pulmão/efeitos da radiação , Masculino , Neutrófilos/efeitos da radiação , Peroxidase/biossíntese , Pneumonia/etiologia , Pneumonia/metabolismo , Pneumonia/patologia , Distribuição Aleatória , Ratos , Ratos WistarRESUMO
It has been suggested that low intensity laser therapy (LILT) acts on pulmonary inflammation. Thus, we investigate in this work if LILT (650nm, 2.5mW, 31.2mW/cm(2), 1.3J/cm(2), laser spot size of 0.08cm(2) and irradiation time of 42s) can attenuate edema, neutrophil recruitment and inflammatory mediators in acute lung inflammation. Thirty-five male Wistar rats (n=7 per group) were distributed in the following experimental groups: control, laser, LPS, LPS+laser and dexamethasone+LPS. Airway inflammation was measured 4h post-LPS challenge. Pulmonary microvascular leakage was used for measuring pulmonary edema. Bronchoalveolar lavage fluid (BALF) cellularity and myeloperoxidase (MPO) were used for measuring neutrophil recruitment and activation. RT-PCR was performed in lung tissue to assess mRNA expression of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin (IL-10), cytokine-induced neutrophil chemoattractant-1 (CINC-1), macrophage inflammatory protein-2 (MIP-2) and intercellular adhesion molecule-1 (ICAM-1). Protein levels in both BALF and lung were determined by ELISA. LILT inhibited pulmonary edema and endothelial cytoskeleton damage, as well as neutrophil influx and activation. Similarly, the LILT reduced the TNF-α and IL-1ß, in lung and BALF. LILT prevented lung ICAM-1 up-regulation. The rise of CINC-1 and MIP-2 protein levels in both lung and BALF, and the lung mRNA expressions for IL-10, were unaffected. Data suggest that the LILT effect is due to the inhibition of ICAM-1 via the inhibition of TNF-α and IL-1ß.
Assuntos
Quimiocinas/metabolismo , Citocinas/metabolismo , Terapia com Luz de Baixa Intensidade , Neutrófilos/efeitos da radiação , Pneumonia/radioterapia , Doença Aguda , Aerossóis/química , Animais , Anti-Inflamatórios/farmacologia , Líquido da Lavagem Broncoalveolar , Quimiocina CXCL1/genética , Quimiocina CXCL1/metabolismo , Quimiocina CXCL2/genética , Quimiocina CXCL2/metabolismo , Quimiocinas/genética , Citocinas/genética , Dexametasona/farmacologia , Modelos Animais de Doenças , Escherichia coli/metabolismo , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/metabolismo , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Lipopolissacarídeos/toxicidade , Pulmão/imunologia , Pulmão/metabolismo , Pulmão/patologia , Masculino , Neutrófilos/metabolismo , Peroxidase/metabolismo , Ratos , Ratos Wistar , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
The mechanism of regeneration does not start to restore the wound until its corresponding epimorphic phase. A bioestimulation of tissues and cells by laser radiation depends on the wavelength, on the dose, and on the intensity of the light. The goal of this work was to verify the effect of the low power laser at 660 nm on the regenerative process of Girardia tigrina. The specimens were maintained in the laboratory under a temperature ranging from 19 degrees up to 24 degrees C for 21 days. The planarians were anesthetized by placing them on ice and then cut them with a scalpel. The three treatments were as following: animals individually irradiated with 14 sessions with 1 minute duration (treatment 1), 14 sessions with 3 minutes duration (treatment 2), and without irradiation (control). The planarians were amputated and divided in three study treatments: a control group (without radiation), and two other treatments: irradiated for 1 minute, and irradiated for 3 minutes. The animals were irradiated with diode laser (660 nm) with 3.3 +/- 0.3 mW of power, using 0.94 mW.mm-2 power density for each irradiation procedure. During the experiment, 14 irradiation sessions were undertaken. The specimens were fixed in Bouin, and stained with hematoxyline and eosin. From observation and histological analysis, it was possible to assess the effects of interaction between laser and tissue. The head fragment after 1 minute of irradiation presented a better organized tissue scheme, when compared with the other treatments. Aspects of the body fragments submitted to 3 minutes of light treatment were very similar to fragments that had not been injured. It can be concluded that there are changes in the quality of regeneration when treated with low power laser under the conditions mentioned above.
Assuntos
Lasers Semicondutores , Terapia com Luz de Baixa Intensidade , Regeneração/efeitos da radiação , Turbelários/efeitos da radiação , Animais , Regeneração/fisiologiaRESUMO
The mechanism of regeneration does not start to restore the wound until its corresponding epimorphic phase. A bioestimulation of tissues and cells by laser radiation depends on the wavelength, on the dose, and on the intensity of the light. The goal of this work was to verify the effect of the low power laser at 660 nm on the regenerative process of Girardia tigrina. The specimens were maintained in the laboratory under a temperature ranging from 19° up to 24 °C for 21 days. The planarians were anesthetized by placing them on ice and then cut them with a scalpel. The three treatments were as following: animals individually irradiated with 14 sessions with 1 minute duration (treatment 1), 14 sessions with 3 minutes duration (treatment 2), and without irradiation (control). The planarians were amputated and divided in three study treatments: a control group (without radiation), and two other treatments: irradiated for 1 minute, and irradiated for 3 minutes. The animals were irradiated with diode laser (660 nm) with 3.3 ± 0.3 mW of power, using 0.94 mW.mm-2 power density for each irradiation procedure. During the experiment, 14 irradiation sessions were undertaken. The specimens were fixed in Bouin, and stained with hematoxyline and eosin. From observation and histological analysis, it was possible to assess the effects of interaction between laser and tissue. The head fragment after 1 minute of irradiation presented a better organized tissue scheme, when compared with the other treatments. Aspects of the body fragments submitted to 3 minutes of light treatment were very similar to fragments that had not been injured. It can be concluded that there are changes in the quality of regeneration when treated with low power laser under the conditions mentioned above.
O mecanismo de regeneração não restaura a ferida até sua fase epimórfica correspondente. A bioestimulação de tecidos e células por radiação laser depende do comprimento de onda, da dose e intensidade de luz. O objetivo deste trabalho foi verificar o efeito do laser de baixa potência (660 nm) no processo regenerativo de Girardia tigrina. Estes vermes têm uma elevada capacidade de regeneração. Os espécimes foram mantidos em laboratório sob uma temperatura de 19° a 24 °C por 21 dias. As planárias foram anestesiadas com gelo e amputadas com bisturi. Os 3 tratamentos foram compostos de: animais individualmente irradiados com 14 sessões de 1 minuto de duração (tratamento 1), com 14 sessões de três minutos de duração (tratamento 2) e não irradiados (controle). Elas foram irradiadas com laser diodo (660 nm) 3,3 ± 0,3 mW potência, usando 0.94 mW.mm-2 de densidade de potência para cada irradiação. Durante o experimento foram realizadas 14 sessões. Os espécimes foram fixados em Bouin e corados com hematoxilina e eosina. Com base nas observações e análise histológica, foi possível avaliar os efeitos da interação entre laser e tecido. O fragmento cabeça após 1 minuto de irradiação mostrou um tecido melhor organizado quando comparado com outros tratamentos. Os aspectos dos fragmentos corpo após 3 minutos de irradiação foram muito similares para os fragmentos que não sofreram injúria. Existem mudanças na qualidade de regeneração quando tratadas com laser nas condições acima mencionadas.
Assuntos
Animais , Lasers Semicondutores , Terapia com Luz de Baixa Intensidade , Regeneração/efeitos da radiação , Turbelários/efeitos da radiação , Regeneração/fisiologiaRESUMO
The mechanism of regeneration does not start to restore the wound until its corresponding epimorphic phase. A bioestimulation of tissues and cells by laser radiation depends on the wavelength, on the dose, and on the intensity of the light. The goal of this work was to verify the effect of the low power laser at 660 nm on the regenerative process of Girardia tigrina. The specimens were maintained in the laboratory under a temperature ranging from 19° up to 24 °C for 21 days. The planarians were anesthetized by placing them on ice and then cut them with a scalpel. The three treatments were as following: animals individually irradiated with 14 sessions with 1 minute duration (treatment 1), 14 sessions with 3 minutes duration (treatment 2), and without irradiation (control). The planarians were amputated and divided in three study treatments: a control group (without radiation), and two other treatments: irradiated for 1 minute, and irradiated for 3 minutes. The animals were irradiated with diode laser (660 nm) with 3.3 ± 0.3 mW of power, using 0.94 mW.mm-2 power density for each irradiation procedure. During the experiment, 14 irradiation sessions were undertaken. The specimens were fixed in Bouin, and stained with hematoxyline and eosin. From observation and histological analysis, it was possible to assess the effects of interaction between laser and tissue. The head fragment after 1 minute of irradiation presented a better organized tissue scheme, when compared with the other treatments. Aspects of the body fragments submitted to 3 minutes of light treatment were very similar to fragments that had not been injured. It can be concluded that there are changes in the quality of regeneration when treated with low power laser under the conditions mentioned above.
O mecanismo de regeneração não restaura a ferida até sua fase epimórfica correspondente. A bioestimulação de tecidos e células por radiação laser depende do comprimento de onda, da dose e intensidade de luz. O objetivo deste trabalho foi verificar o efeito do laser de baixa potência (660 nm) no processo regenerativo de Girardia tigrina. Estes vermes têm uma elevada capacidade de regeneração. Os espécimes foram mantidos em laboratório sob uma temperatura de 19° a 24 °C por 21 dias. As planárias foram anestesiadas com gelo e amputadas com bisturi. Os 3 tratamentos foram compostos de: animais individualmente irradiados com 14 sessões de 1 minuto de duração (tratamento 1), com 14 sessões de três minutos de duração (tratamento 2) e não irradiados (controle). Elas foram irradiadas com laser diodo (660 nm) 3,3 ± 0,3 mW potência, usando 0.94 mW.mm-2 de densidade de potência para cada irradiação. Durante o experimento foram realizadas 14 sessões. Os espécimes foram fixados em Bouin e corados com hematoxilina e eosina. Com base nas observações e análise histológica, foi possível avaliar os efeitos da interação entre laser e tecido. O fragmento cabeça após 1 minuto de irradiação mostrou um tecido melhor organizado quando comparado com outros tratamentos. Os aspectos dos fragmentos corpo após 3 minutos de irradiação foram muito similares para os fragmentos que não sofreram injúria. Existem mudanças na qualidade de regeneração quando tratadas com laser nas condições acima mencionadas.
RESUMO
It has been suggested that low-level laser therapy (LLLT) can modulate inflammatory processes. The aim of this experiment was to investigate what effects red laser irradiation with two different wavelengths (660 nm and 684 nm) on carrageenan-induced rat paw edema and histology. Thirty two male Wistar rats were randomly divided into four groups. One group received a sterile saline injection, while inflammation was induced by a sub-plantar injection of carrageenan (1 mg/paw) in the three other groups. After 1 h, LLLT was administered to the paw in two of the carrageenan-injected groups. Continuous wave 660 nm and 684 nm red lasers respectively with mean optical outputs of 30 mW and doses of 7.5 J/cm(2) were used. The 660 nm and 684 nm laser groups developed significantly (p<0.01) less edema (0.58 ml [SE+/-0.17] ml and 0.76 ml [SE+/-0.10] respectively) than the control group (1.67 ml [SE+/-0.19]) at 4h after injections. Similarly, both laser groups showed a significantly lower number of inflammatory cells in the muscular and conjunctive sub-plantar tissues than the control group. We conclude that both 660 nm and 684 nm red wavelengths of LLLT are effective in reducing edema formation and inflammatory cell migration when a dose of 7.5 J/cm(2) is used.