Photobiomodulation assay of muscle cells C2C12 after irradiation with LED device

Detalhes bibliográficos
Autor(a) principal: Lima, Elessandro Váguino de
Data de Publicação: 2022
Outros Autores: Pacheco-Soares, Cristina, Silva, Newton Soares da
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Research, Society and Development
Texto Completo: https://rsdjournal.org/index.php/rsd/article/view/28884
Resumo: Introduction: One of the ways that have been observed to reduce musculoskeletal fatigue is the use of protocols for the application of light sources (photobiomodulation) such as low-intensity laser and LED (Light Emitting Diode). Work involving photobiomodulation has shown promising results in strength performance or reduction of muscle fatigue. At the cellular level, photobiomodulation can modulate fibroblasts proliferation, the fixation and synthesis of collagen and procollagen, promote angiogenesis and improving energy metabolism in mitochondria. Compared with laser devices, LED has several advantages, such as being smaller, lighter, lower cost, and easier for operation. Objective: The present work objective is to verify if irradiation with LED device (650 nm and 860 nm) in muscle cells C2C12 modify the viability, morphology and cytoskeleton components. Methodology: C2C12 cells line (ATCC CRL - 1772) were cultured in 25 cm2 bottles at 37ºC under 5% CO2 in DMEM. The cells were irradiated with the light-emitting diodes (LED) device, Sportllux Ultra that consists of 84 LEDs, each individual LED has 8 mW of power, emitting in 660±20 nm (42 LEDs) and 850±20 nm (42 LEDs), and covering an area (A) of 120 cm2. The power density of delivered light was 5,6 mW/cm2, and the exposure time was 10 minutes, totalizing the fluence of 3,4 J/cm2. Viability assay was performed where the cells were incubated with 100 µL of Crystal Violet (CV) solution and mitochondrial activity assay was evaluated by the colorimetric MTT assay. Nucleus (DAPI) and Cytoskeleton (Rhodamine Phalloidin) fluorescence assay was performed to study the cytoskeleton based on the change in the actin filaments. Results: Our results demonstrate that the synergism of LED irradiation (660nm and 850nm) induced the proliferation of C2C12 cells. The light-emitting diode (LED) device, Sportllux Ultra has a significant effect on C2C12 cell culture. Mitochondrial activity and cell viability showed a significative increase in their activities after irradiation. The microscopy fluorescence observations showed an alignment of cytoskeletal components of C2C12 cells after irradiation. Conclusion: The application of irradiation with the Sportlux Ultra LED device stimulated an increase of energy by mitochondrial activity assay, number of cells by cell viability assay and alignment of cytoskeleton components by fluorescence assay in C2C12 line cells. Our results suggest that organizated cytoskeletal actin filaments normally contribute to cell survival and that induced major cell changes in the cytoskeleton that result in cell shape change. These results suggest that the Sport Lux Ultra LED device can help in the repair of tissue injuries and to collaborate to increase of performance in athletes in a faster way.
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spelling Photobiomodulation assay of muscle cells C2C12 after irradiation with LED deviceEnsayo de fotobiomodulación de células musculares C2C12 después de la irradiación con dispositivo LEDEnsaio de fotobiomodulação de células musculares C2C12 após irradiação com dispositivo LEDFotobiomodulaçãoCultura de célulasCélulas musculares.FotobiomodulaciónCultivo de célulasCélulas musculares.PhotobiomodulationCell cultureMuscle cells.Introduction: One of the ways that have been observed to reduce musculoskeletal fatigue is the use of protocols for the application of light sources (photobiomodulation) such as low-intensity laser and LED (Light Emitting Diode). Work involving photobiomodulation has shown promising results in strength performance or reduction of muscle fatigue. At the cellular level, photobiomodulation can modulate fibroblasts proliferation, the fixation and synthesis of collagen and procollagen, promote angiogenesis and improving energy metabolism in mitochondria. Compared with laser devices, LED has several advantages, such as being smaller, lighter, lower cost, and easier for operation. Objective: The present work objective is to verify if irradiation with LED device (650 nm and 860 nm) in muscle cells C2C12 modify the viability, morphology and cytoskeleton components. Methodology: C2C12 cells line (ATCC CRL - 1772) were cultured in 25 cm2 bottles at 37ºC under 5% CO2 in DMEM. The cells were irradiated with the light-emitting diodes (LED) device, Sportllux Ultra that consists of 84 LEDs, each individual LED has 8 mW of power, emitting in 660±20 nm (42 LEDs) and 850±20 nm (42 LEDs), and covering an area (A) of 120 cm2. The power density of delivered light was 5,6 mW/cm2, and the exposure time was 10 minutes, totalizing the fluence of 3,4 J/cm2. Viability assay was performed where the cells were incubated with 100 µL of Crystal Violet (CV) solution and mitochondrial activity assay was evaluated by the colorimetric MTT assay. Nucleus (DAPI) and Cytoskeleton (Rhodamine Phalloidin) fluorescence assay was performed to study the cytoskeleton based on the change in the actin filaments. Results: Our results demonstrate that the synergism of LED irradiation (660nm and 850nm) induced the proliferation of C2C12 cells. The light-emitting diode (LED) device, Sportllux Ultra has a significant effect on C2C12 cell culture. Mitochondrial activity and cell viability showed a significative increase in their activities after irradiation. The microscopy fluorescence observations showed an alignment of cytoskeletal components of C2C12 cells after irradiation. Conclusion: The application of irradiation with the Sportlux Ultra LED device stimulated an increase of energy by mitochondrial activity assay, number of cells by cell viability assay and alignment of cytoskeleton components by fluorescence assay in C2C12 line cells. Our results suggest that organizated cytoskeletal actin filaments normally contribute to cell survival and that induced major cell changes in the cytoskeleton that result in cell shape change. These results suggest that the Sport Lux Ultra LED device can help in the repair of tissue injuries and to collaborate to increase of performance in athletes in a faster way.Introducción: Una de las formas observadas para reducir la fatiga musculoesquelética es el uso de protocolos para la aplicación de fuentes de luz (fotobiomodulación) como láser de baja intensidad y LED (Light Emitting Diode). Los trabajos de fotobiomodulación han mostrado resultados prometedores en el rendimiento de la fuerza o la reducción de la fatiga muscular. A nivel celular, la fotobiomodulación puede modular la proliferación de fibroblastos, la unión y síntesis de colágeno y procolágeno, promover la angiogénesis y mejorar el metabolismo energético en las mitocondrias. En comparación con los dispositivos láser, el LED tiene varias ventajas, como ser más pequeño, más liviano, menos costoso y más fácil de operar. Objetivo: El objetivo del presente trabajo es verificar si la irradiación con dispositivo LED (650 nm y 860 nm) en células musculares C2C12 modifica la viabilidad, morfología y componentes del citoesqueleto. Metodología: La línea celular C2C12 (ATCC CRL - 1772) se cultivó en matraces de 25 cm2 a 37°C bajo 5% de CO2 en DMEM. Las celdas fueron irradiadas con el dispositivo de diodo emisor de luz (LED), Sportllux Ultra, que consta de 84 LED, cada LED individual tiene 8 mW de potencia, emitiendo a 660±20 nm (42 LED) y 850±20 nm (42 LEDs), y cubriendo un área (A) de 120 cm2. La densidad de potencia de la luz emitida fue de 5,6 mW/cm2 y el tiempo de exposición de 10 minutos, totalizando una fluencia de 3,4 J/cm2. El ensayo de viabilidad se realizó donde las células se incubaron con 100 µL de solución Crystal Violet (CV) y el ensayo de actividad mitocondrial se evaluó mediante el ensayo colorimétrico MTT. Se realizó el ensayo de núcleo de fluorescencia (DAPI) y citoesqueleto (faloidina de rodamina) para estudiar el citoesqueleto en función del cambio en los filamentos de actina. Resultados: Nuestros resultados demuestran que el sinergismo de la irradiación LED (660nm y 850nm) indujo la proliferación de células C2C12. El dispositivo de diodo emisor de luz (LED), Sportllux Ultra tiene un efecto significativo en el cultivo de células C2C12. La actividad mitocondrial y la viabilidad celular mostraron un aumento significativo en sus actividades después de la irradiación. Las observaciones de microscopía de fluorescencia mostraron una alineación de los componentes del citoesqueleto de las células C2C12 después de la irradiación. Conclusión: La aplicación de irradiación con el dispositivo Sportlux Ultra LED estimuló un aumento de energía por ensayo de actividad mitocondrial, número de células por ensayo de viabilidad celular y alineación de componentes del citoesqueleto por ensayo de fluorescencia en células de la línea C2C12. Nuestros resultados sugieren que los filamentos de actina del citoesqueleto organizados normalmente contribuyen a la supervivencia celular y que indujeron cambios celulares importantes en el citoesqueleto que dan como resultado un cambio en la forma de la célula. Estos resultados sugieren que el dispositivo Sport Lux Ultra LED puede ayudar en la reparación de lesiones en los tejidos y colaborar para aumentar el rendimiento en los atletas de una manera más rápida.Introdução: Uma das formas observadas para a redução da fadiga musculoesquelética é a utilização de protocolos para aplicação de fontes de luz (fotobiomodulação) como laser de baixa intensidade e LED (Light Emitting Diode). Trabalhos envolvendo fotobiomodulação têm mostrado resultados promissores no desempenho de força ou redução da fadiga muscular. Em nível celular, a fotobiomodulação pode modular a proliferação de fibroblastos, a fixação e síntese de colágeno e procolágeno, promover a angiogênese e melhorar o metabolismo energético nas mitocôndrias. Comparado aos dispositivos a laser, o LED apresenta diversas vantagens, como ser menor, mais leve, de menor custo e de operação mais fácil. Objetivo: O objetivo do presente trabalho é verificar se a irradiação com dispositivo LED (650 nm e 860 nm) em células musculares C2C12 modifica a viabilidade, morfologia e componentes do citoesqueleto. Metodologia: A linhagem celular C2C12 (ATCC CRL - 1772) foi cultivada em frascos de 25 cm2 a 37ºC sob 5% CO2 em DMEM. As células foram irradiadas com o dispositivo de diodos emissores de luz (LED), Sportllux Ultra que consiste em 84 LEDs, cada LED individual possui 8 mW de potência, emitindo em 660±20 nm (42 LEDs) e 850±20 nm (42 LEDs), e cobrindo uma área (A) de 120 cm2. A densidade de potência da luz emitida foi de 5,6 mW/cm2, e o tempo de exposição foi de 10 minutos, totalizando a fluência de 3,4 J/cm2. O ensaio de viabilidade foi realizado onde as células foram incubadas com 100 µL de solução de Cristal Violeta (CV) e o ensaio de atividade mitocondrial foi avaliado pelo ensaio colorimétrico MTT. O ensaio de fluorescência de núcleo (DAPI) e citoesqueleto (rodamina faloidina) foi realizado para estudar o citoesqueleto com base na alteração nos filamentos de actina. Resultados: Nossos resultados demonstram que o sinergismo da irradiação LED (660nm e 850nm) induziu a proliferação de células C2C12. O dispositivo de diodo emissor de luz (LED), Sportllux Ultra tem um efeito significativo na cultura de células C2C12. A atividade mitocondrial e a viabilidade celular mostraram um aumento significativo em suas atividades após a irradiação. As observações de microscopia de fluorescência mostraram um alinhamento dos componentes do citoesqueleto das células C2C12 após a irradiação. Conclusão: A aplicação da irradiação com o aparelho Sportlux Ultra LED estimulou aumento de energia por ensaio de atividade mitocondrial, número de células por ensaio de viabilidade celular e alinhamento de componentes do citoesqueleto por ensaio de fluorescência em células da linhagem C2C12. Nossos resultados sugerem que os filamentos organizados de actina do citoesqueleto normalmente contribuem para a sobrevivência da célula e que induzem grandes mudanças celulares no citoesqueleto que resultam na mudança da forma celular. Esses resultados sugerem que o aparelho Sport Lux Ultra LED pode auxiliar no reparo de lesões teciduais e colaborar para aumentar o desempenho em atletas de forma mais rápida.Research, Society and Development2022-05-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://rsdjournal.org/index.php/rsd/article/view/2888410.33448/rsd-v11i6.28884Research, Society and Development; Vol. 11 No. 6; e41511628884Research, Society and Development; Vol. 11 Núm. 6; e41511628884Research, Society and Development; v. 11 n. 6; e415116288842525-3409reponame:Research, Society and Developmentinstname:Universidade Federal de Itajubá (UNIFEI)instacron:UNIFEIenghttps://rsdjournal.org/index.php/rsd/article/view/28884/25353Copyright (c) 2022 Elessandro Váguino de Lima; Cristina Pacheco-Soares; Newton Soares da Silvahttps://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessLima, Elessandro Váguino de Pacheco-Soares, CristinaSilva, Newton Soares da 2022-05-13T18:04:10Zoai:ojs.pkp.sfu.ca:article/28884Revistahttps://rsdjournal.org/index.php/rsd/indexPUBhttps://rsdjournal.org/index.php/rsd/oairsd.articles@gmail.com2525-34092525-3409opendoar:2024-01-17T09:46:08.320503Research, Society and Development - Universidade Federal de Itajubá (UNIFEI)false
dc.title.none.fl_str_mv Photobiomodulation assay of muscle cells C2C12 after irradiation with LED device
Ensayo de fotobiomodulación de células musculares C2C12 después de la irradiación con dispositivo LED
Ensaio de fotobiomodulação de células musculares C2C12 após irradiação com dispositivo LED
title Photobiomodulation assay of muscle cells C2C12 after irradiation with LED device
spellingShingle Photobiomodulation assay of muscle cells C2C12 after irradiation with LED device
Lima, Elessandro Váguino de
Fotobiomodulação
Cultura de células
Células musculares.
Fotobiomodulación
Cultivo de células
Células musculares.
Photobiomodulation
Cell culture
Muscle cells.
title_short Photobiomodulation assay of muscle cells C2C12 after irradiation with LED device
title_full Photobiomodulation assay of muscle cells C2C12 after irradiation with LED device
title_fullStr Photobiomodulation assay of muscle cells C2C12 after irradiation with LED device
title_full_unstemmed Photobiomodulation assay of muscle cells C2C12 after irradiation with LED device
title_sort Photobiomodulation assay of muscle cells C2C12 after irradiation with LED device
author Lima, Elessandro Váguino de
author_facet Lima, Elessandro Váguino de
Pacheco-Soares, Cristina
Silva, Newton Soares da
author_role author
author2 Pacheco-Soares, Cristina
Silva, Newton Soares da
author2_role author
author
dc.contributor.author.fl_str_mv Lima, Elessandro Váguino de
Pacheco-Soares, Cristina
Silva, Newton Soares da
dc.subject.por.fl_str_mv Fotobiomodulação
Cultura de células
Células musculares.
Fotobiomodulación
Cultivo de células
Células musculares.
Photobiomodulation
Cell culture
Muscle cells.
topic Fotobiomodulação
Cultura de células
Células musculares.
Fotobiomodulación
Cultivo de células
Células musculares.
Photobiomodulation
Cell culture
Muscle cells.
description Introduction: One of the ways that have been observed to reduce musculoskeletal fatigue is the use of protocols for the application of light sources (photobiomodulation) such as low-intensity laser and LED (Light Emitting Diode). Work involving photobiomodulation has shown promising results in strength performance or reduction of muscle fatigue. At the cellular level, photobiomodulation can modulate fibroblasts proliferation, the fixation and synthesis of collagen and procollagen, promote angiogenesis and improving energy metabolism in mitochondria. Compared with laser devices, LED has several advantages, such as being smaller, lighter, lower cost, and easier for operation. Objective: The present work objective is to verify if irradiation with LED device (650 nm and 860 nm) in muscle cells C2C12 modify the viability, morphology and cytoskeleton components. Methodology: C2C12 cells line (ATCC CRL - 1772) were cultured in 25 cm2 bottles at 37ºC under 5% CO2 in DMEM. The cells were irradiated with the light-emitting diodes (LED) device, Sportllux Ultra that consists of 84 LEDs, each individual LED has 8 mW of power, emitting in 660±20 nm (42 LEDs) and 850±20 nm (42 LEDs), and covering an area (A) of 120 cm2. The power density of delivered light was 5,6 mW/cm2, and the exposure time was 10 minutes, totalizing the fluence of 3,4 J/cm2. Viability assay was performed where the cells were incubated with 100 µL of Crystal Violet (CV) solution and mitochondrial activity assay was evaluated by the colorimetric MTT assay. Nucleus (DAPI) and Cytoskeleton (Rhodamine Phalloidin) fluorescence assay was performed to study the cytoskeleton based on the change in the actin filaments. Results: Our results demonstrate that the synergism of LED irradiation (660nm and 850nm) induced the proliferation of C2C12 cells. The light-emitting diode (LED) device, Sportllux Ultra has a significant effect on C2C12 cell culture. Mitochondrial activity and cell viability showed a significative increase in their activities after irradiation. The microscopy fluorescence observations showed an alignment of cytoskeletal components of C2C12 cells after irradiation. Conclusion: The application of irradiation with the Sportlux Ultra LED device stimulated an increase of energy by mitochondrial activity assay, number of cells by cell viability assay and alignment of cytoskeleton components by fluorescence assay in C2C12 line cells. Our results suggest that organizated cytoskeletal actin filaments normally contribute to cell survival and that induced major cell changes in the cytoskeleton that result in cell shape change. These results suggest that the Sport Lux Ultra LED device can help in the repair of tissue injuries and to collaborate to increase of performance in athletes in a faster way.
publishDate 2022
dc.date.none.fl_str_mv 2022-05-02
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
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dc.identifier.uri.fl_str_mv https://rsdjournal.org/index.php/rsd/article/view/28884
10.33448/rsd-v11i6.28884
url https://rsdjournal.org/index.php/rsd/article/view/28884
identifier_str_mv 10.33448/rsd-v11i6.28884
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv https://rsdjournal.org/index.php/rsd/article/view/28884/25353
dc.rights.driver.fl_str_mv Copyright (c) 2022 Elessandro Váguino de Lima; Cristina Pacheco-Soares; Newton Soares da Silva
https://creativecommons.org/licenses/by/4.0
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Copyright (c) 2022 Elessandro Váguino de Lima; Cristina Pacheco-Soares; Newton Soares da Silva
https://creativecommons.org/licenses/by/4.0
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Research, Society and Development
publisher.none.fl_str_mv Research, Society and Development
dc.source.none.fl_str_mv Research, Society and Development; Vol. 11 No. 6; e41511628884
Research, Society and Development; Vol. 11 Núm. 6; e41511628884
Research, Society and Development; v. 11 n. 6; e41511628884
2525-3409
reponame:Research, Society and Development
instname:Universidade Federal de Itajubá (UNIFEI)
instacron:UNIFEI
instname_str Universidade Federal de Itajubá (UNIFEI)
instacron_str UNIFEI
institution UNIFEI
reponame_str Research, Society and Development
collection Research, Society and Development
repository.name.fl_str_mv Research, Society and Development - Universidade Federal de Itajubá (UNIFEI)
repository.mail.fl_str_mv rsd.articles@gmail.com
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