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Investigating the effects of low intensity visible light on human keratinocytes using a customized LED exposure system
Photobiomodulation (PBM) refers to the use of light to modulate cellular processes, and has demonstrated utility in improving wound healing outcomes, and reducing pain and inflammation. Despite the potential benefits of PBM, the precise molecular mechanisms through which it influences cell behavior...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9640685/ https://www.ncbi.nlm.nih.gov/pubmed/36344673 http://dx.doi.org/10.1038/s41598-022-23751-3 |
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author | Sutterby, Emily Chheang, Chanly Thurgood, Peter Khoshmanesh, Khashayar Baratchi, Sara Pirogova, Elena |
author_facet | Sutterby, Emily Chheang, Chanly Thurgood, Peter Khoshmanesh, Khashayar Baratchi, Sara Pirogova, Elena |
author_sort | Sutterby, Emily |
collection | PubMed |
description | Photobiomodulation (PBM) refers to the use of light to modulate cellular processes, and has demonstrated utility in improving wound healing outcomes, and reducing pain and inflammation. Despite the potential benefits of PBM, the precise molecular mechanisms through which it influences cell behavior are not yet well understood. Inconsistent reporting of key light parameters has created uncertainty around optimal exposure profiles. In addition, very low intensities of light, < 0.1 J/cm(2), have not been thoroughly examined for their use in PBM. Here, we present a custom-made compact, and modular LED-based exposure system for studying the effects of very low-intensity visible light (cell proliferation, migration, ROS production, and mitochondrial membrane potential) of three different wavelengths in a parallel manner. The device allows for six repeats of three different exposure conditions plus a non-irradiated control on a single 24-well plate. The immortalised human keratinocyte cell line, HaCaT, was selected as a major cellular component of the skin epidermal barrier. Furthermore, an in vitro wound model was developed by allowing the HaCaT to form a confluent monolayer, then scratching the cells with a pipette tip to form a wound. Cells were exposed to yellow (585 nm, 0.09 mW, ~ 3.7 mJ/cm(2)), orange (610 nm, 0.8 mW, ~ 31 mJ/cm(2)), and red (660 nm, 0.8 mW, ~ 31 mJ/cm(2)) light for 10 min. 48 h post-irradiation, immunohistochemistry was performed to evaluate cell viability, proliferation, ROS production, and mitochondrial membrane potential. The results demonstrate increased proliferation and decreased scratch area for all exposure conditions, however only red light increased the mitochondrial activity. Oxidative stress levels did not increase for any of the exposures. The present exposure system provides opportunities to better understand the complex cellular mechanisms driven by the irradiation of skin cells with visible light. |
format | Online Article Text |
id | pubmed-9640685 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-96406852022-11-15 Investigating the effects of low intensity visible light on human keratinocytes using a customized LED exposure system Sutterby, Emily Chheang, Chanly Thurgood, Peter Khoshmanesh, Khashayar Baratchi, Sara Pirogova, Elena Sci Rep Article Photobiomodulation (PBM) refers to the use of light to modulate cellular processes, and has demonstrated utility in improving wound healing outcomes, and reducing pain and inflammation. Despite the potential benefits of PBM, the precise molecular mechanisms through which it influences cell behavior are not yet well understood. Inconsistent reporting of key light parameters has created uncertainty around optimal exposure profiles. In addition, very low intensities of light, < 0.1 J/cm(2), have not been thoroughly examined for their use in PBM. Here, we present a custom-made compact, and modular LED-based exposure system for studying the effects of very low-intensity visible light (cell proliferation, migration, ROS production, and mitochondrial membrane potential) of three different wavelengths in a parallel manner. The device allows for six repeats of three different exposure conditions plus a non-irradiated control on a single 24-well plate. The immortalised human keratinocyte cell line, HaCaT, was selected as a major cellular component of the skin epidermal barrier. Furthermore, an in vitro wound model was developed by allowing the HaCaT to form a confluent monolayer, then scratching the cells with a pipette tip to form a wound. Cells were exposed to yellow (585 nm, 0.09 mW, ~ 3.7 mJ/cm(2)), orange (610 nm, 0.8 mW, ~ 31 mJ/cm(2)), and red (660 nm, 0.8 mW, ~ 31 mJ/cm(2)) light for 10 min. 48 h post-irradiation, immunohistochemistry was performed to evaluate cell viability, proliferation, ROS production, and mitochondrial membrane potential. The results demonstrate increased proliferation and decreased scratch area for all exposure conditions, however only red light increased the mitochondrial activity. Oxidative stress levels did not increase for any of the exposures. The present exposure system provides opportunities to better understand the complex cellular mechanisms driven by the irradiation of skin cells with visible light. Nature Publishing Group UK 2022-11-07 /pmc/articles/PMC9640685/ /pubmed/36344673 http://dx.doi.org/10.1038/s41598-022-23751-3 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Sutterby, Emily Chheang, Chanly Thurgood, Peter Khoshmanesh, Khashayar Baratchi, Sara Pirogova, Elena Investigating the effects of low intensity visible light on human keratinocytes using a customized LED exposure system |
title | Investigating the effects of low intensity visible light on human keratinocytes using a customized LED exposure system |
title_full | Investigating the effects of low intensity visible light on human keratinocytes using a customized LED exposure system |
title_fullStr | Investigating the effects of low intensity visible light on human keratinocytes using a customized LED exposure system |
title_full_unstemmed | Investigating the effects of low intensity visible light on human keratinocytes using a customized LED exposure system |
title_short | Investigating the effects of low intensity visible light on human keratinocytes using a customized LED exposure system |
title_sort | investigating the effects of low intensity visible light on human keratinocytes using a customized led exposure system |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9640685/ https://www.ncbi.nlm.nih.gov/pubmed/36344673 http://dx.doi.org/10.1038/s41598-022-23751-3 |
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