Skeletal cell differentiation is enhanced by atmospheric dielectric barrier discharge plasma treatment
Enhancing chondrogenic and osteogenic differentiation is of paramount importance in providing effective regenerative therapies and improving the rate of fracture healing. This study investigated the potential of non-thermal atmospheric dielectric barrier discharge plasma (NT-plasma) to enhance chond...
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| creator | Steinbeck, Marla J Chernets, Natalie Zhang, Jun Kurpad, Deepa S Fridman, Gregory Fridman, Alexander Freeman, Theresa A |
| description | Enhancing chondrogenic and osteogenic differentiation is of paramount importance in providing effective regenerative therapies and improving the rate of fracture healing. This study investigated the potential of non-thermal atmospheric dielectric barrier discharge plasma (NT-plasma) to enhance chondrocyte and osteoblast proliferation and differentiation. Although the exact mechanism by which NT-plasma interacts with cells is undefined, it is known that during treatment the atmosphere is ionized generating extracellular reactive oxygen and nitrogen species (ROS and RNS) and an electric field. Appropriate NT-plasma conditions were determined using lactate-dehydrogenase release, flow cytometric live/dead assay, flow cytometric cell cycle analysis, and Western blots to evaluate DNA damage and mitochondrial integrity. We observed that specific NT-plasma conditions were required to prevent cell death, and that loss of pre-osteoblastic cell viability was dependent on intracellular ROS and RNS production. To further investigate the involvement of intracellular ROS, fluorescent intracellular dyes Mitosox (superoxide) and dihydrorhodamine (peroxide) were used to assess onset and duration after NT-plasma treatment. Both intracellular superoxide and peroxide were found to increase immediately post NT-plasma treatment. These increases were sustained for one hour but returned to control levels by 24 hr. Using the same treatment conditions, osteogenic differentiation by NT-plasma was assessed and compared to peroxide or osteogenic media containing β-glycerolphosphate. Although both NT-plasma and peroxide induced differentiation-specific gene expression, neither was as effective as the osteogenic media. However, treatment of cells with NT-plasma after 24 hr in osteogenic or chondrogenic media significantly enhanced differentiation as compared to differentiation media alone. The results of this study show that NT-plasma can selectively initiate and amplify ROS signaling to enhance differentiation, and suggest this technology could be used to enhance bone fusion and improve healing after skeletal injury. |
| doi_str_mv | 10.1371/journal.pone.0082143 |
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This study investigated the potential of non-thermal atmospheric dielectric barrier discharge plasma (NT-plasma) to enhance chondrocyte and osteoblast proliferation and differentiation. Although the exact mechanism by which NT-plasma interacts with cells is undefined, it is known that during treatment the atmosphere is ionized generating extracellular reactive oxygen and nitrogen species (ROS and RNS) and an electric field. Appropriate NT-plasma conditions were determined using lactate-dehydrogenase release, flow cytometric live/dead assay, flow cytometric cell cycle analysis, and Western blots to evaluate DNA damage and mitochondrial integrity. We observed that specific NT-plasma conditions were required to prevent cell death, and that loss of pre-osteoblastic cell viability was dependent on intracellular ROS and RNS production. To further investigate the involvement of intracellular ROS, fluorescent intracellular dyes Mitosox (superoxide) and dihydrorhodamine (peroxide) were used to assess onset and duration after NT-plasma treatment. Both intracellular superoxide and peroxide were found to increase immediately post NT-plasma treatment. These increases were sustained for one hour but returned to control levels by 24 hr. Using the same treatment conditions, osteogenic differentiation by NT-plasma was assessed and compared to peroxide or osteogenic media containing β-glycerolphosphate. Although both NT-plasma and peroxide induced differentiation-specific gene expression, neither was as effective as the osteogenic media. However, treatment of cells with NT-plasma after 24 hr in osteogenic or chondrogenic media significantly enhanced differentiation as compared to differentiation media alone. The results of this study show that NT-plasma can selectively initiate and amplify ROS signaling to enhance differentiation, and suggest this technology could be used to enhance bone fusion and improve healing after skeletal injury.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0082143</identifier><identifier>PMID: 24349203</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Analysis ; Animals ; Apoptosis ; Atmosphere - chemistry ; Biocompatibility ; Biomedical engineering ; Biomedical materials ; Bone healing ; Cell cycle ; Cell death ; Cell Death - drug effects ; Cell differentiation ; Cell Differentiation - drug effects ; Cell Line ; Cell Proliferation - drug effects ; Chondrocytes ; Chondrocytes - cytology ; Chondrocytes - drug effects ; Chondrocytes - metabolism ; Chondrogenesis ; Damage assessment ; Deoxyribonucleic acid ; Dielectric barrier discharge ; Differentiation (biology) ; DNA ; DNA damage ; Electricity ; Electrodes ; Engineering ; Flow cytometry ; Fluorescence ; Gene expression ; Growth factors ; Healing ; High temperature ; Intracellular ; Intracellular Space - metabolism ; Kinases ; Lactic acid ; Mice ; Mitochondrial DNA ; Nitric oxide ; Osteoblastogenesis ; Osteoblasts ; Osteoblasts - cytology ; Osteoblasts - drug effects ; Osteoblasts - metabolism ; Osteogenesis - drug effects ; Oxygen ; Penicillin ; Peroxide ; Plasma ; Plasma Gases - pharmacology ; Plasma physics ; Polymerase Chain Reaction ; Reactive nitrogen species ; Reactive oxygen species ; Reactive Oxygen Species - metabolism ; Signaling ; Superoxide ; Superoxides ; Surgery ; Western blotting</subject><ispartof>PloS one, 2013-12, Vol.8 (12), p.e82143-e82143</ispartof><rights>COPYRIGHT 2013 Public Library of Science</rights><rights>2013 Steinbeck et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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This study investigated the potential of non-thermal atmospheric dielectric barrier discharge plasma (NT-plasma) to enhance chondrocyte and osteoblast proliferation and differentiation. Although the exact mechanism by which NT-plasma interacts with cells is undefined, it is known that during treatment the atmosphere is ionized generating extracellular reactive oxygen and nitrogen species (ROS and RNS) and an electric field. Appropriate NT-plasma conditions were determined using lactate-dehydrogenase release, flow cytometric live/dead assay, flow cytometric cell cycle analysis, and Western blots to evaluate DNA damage and mitochondrial integrity. We observed that specific NT-plasma conditions were required to prevent cell death, and that loss of pre-osteoblastic cell viability was dependent on intracellular ROS and RNS production. To further investigate the involvement of intracellular ROS, fluorescent intracellular dyes Mitosox (superoxide) and dihydrorhodamine (peroxide) were used to assess onset and duration after NT-plasma treatment. Both intracellular superoxide and peroxide were found to increase immediately post NT-plasma treatment. These increases were sustained for one hour but returned to control levels by 24 hr. Using the same treatment conditions, osteogenic differentiation by NT-plasma was assessed and compared to peroxide or osteogenic media containing β-glycerolphosphate. Although both NT-plasma and peroxide induced differentiation-specific gene expression, neither was as effective as the osteogenic media. However, treatment of cells with NT-plasma after 24 hr in osteogenic or chondrogenic media significantly enhanced differentiation as compared to differentiation media alone. 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Steinbeck, Marla J</au><au>Chernets, Natalie</au><au>Zhang, Jun</au><au>Kurpad, Deepa S</au><au>Fridman, Gregory</au><au>Fridman, Alexander</au><au>Freeman, Theresa A</au><au>Yamamoto, Masaya</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Skeletal cell differentiation is enhanced by atmospheric dielectric barrier discharge plasma treatment</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2013-12-12</date><risdate>2013</risdate><volume>8</volume><issue>12</issue><spage>e82143</spage><epage>e82143</epage><pages>e82143-e82143</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Enhancing chondrogenic and osteogenic differentiation is of paramount importance in providing effective regenerative therapies and improving the rate of fracture healing. This study investigated the potential of non-thermal atmospheric dielectric barrier discharge plasma (NT-plasma) to enhance chondrocyte and osteoblast proliferation and differentiation. Although the exact mechanism by which NT-plasma interacts with cells is undefined, it is known that during treatment the atmosphere is ionized generating extracellular reactive oxygen and nitrogen species (ROS and RNS) and an electric field. Appropriate NT-plasma conditions were determined using lactate-dehydrogenase release, flow cytometric live/dead assay, flow cytometric cell cycle analysis, and Western blots to evaluate DNA damage and mitochondrial integrity. We observed that specific NT-plasma conditions were required to prevent cell death, and that loss of pre-osteoblastic cell viability was dependent on intracellular ROS and RNS production. To further investigate the involvement of intracellular ROS, fluorescent intracellular dyes Mitosox (superoxide) and dihydrorhodamine (peroxide) were used to assess onset and duration after NT-plasma treatment. Both intracellular superoxide and peroxide were found to increase immediately post NT-plasma treatment. These increases were sustained for one hour but returned to control levels by 24 hr. Using the same treatment conditions, osteogenic differentiation by NT-plasma was assessed and compared to peroxide or osteogenic media containing β-glycerolphosphate. Although both NT-plasma and peroxide induced differentiation-specific gene expression, neither was as effective as the osteogenic media. However, treatment of cells with NT-plasma after 24 hr in osteogenic or chondrogenic media significantly enhanced differentiation as compared to differentiation media alone. The results of this study show that NT-plasma can selectively initiate and amplify ROS signaling to enhance differentiation, and suggest this technology could be used to enhance bone fusion and improve healing after skeletal injury.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24349203</pmid><doi>10.1371/journal.pone.0082143</doi><tpages>e82143</tpages><oa>free_for_read</oa></addata></record> |
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| recordid | cdi_plos_journals_1467678314 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Analysis Animals Apoptosis Atmosphere - chemistry Biocompatibility Biomedical engineering Biomedical materials Bone healing Cell cycle Cell death Cell Death - drug effects Cell differentiation Cell Differentiation - drug effects Cell Line Cell Proliferation - drug effects Chondrocytes Chondrocytes - cytology Chondrocytes - drug effects Chondrocytes - metabolism Chondrogenesis Damage assessment Deoxyribonucleic acid Dielectric barrier discharge Differentiation (biology) DNA DNA damage Electricity Electrodes Engineering Flow cytometry Fluorescence Gene expression Growth factors Healing High temperature Intracellular Intracellular Space - metabolism Kinases Lactic acid Mice Mitochondrial DNA Nitric oxide Osteoblastogenesis Osteoblasts Osteoblasts - cytology Osteoblasts - drug effects Osteoblasts - metabolism Osteogenesis - drug effects Oxygen Penicillin Peroxide Plasma Plasma Gases - pharmacology Plasma physics Polymerase Chain Reaction Reactive nitrogen species Reactive oxygen species Reactive Oxygen Species - metabolism Signaling Superoxide Superoxides Surgery Western blotting |
| title | Skeletal cell differentiation is enhanced by atmospheric dielectric barrier discharge plasma treatment |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-27T08%3A20%3A28IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Skeletal%20cell%20differentiation%20is%20enhanced%20by%20atmospheric%20dielectric%20barrier%20discharge%20plasma%20treatment&rft.jtitle=PloS%20one&rft.au=Steinbeck,%20Marla%20J&rft.date=2013-12-12&rft.volume=8&rft.issue=12&rft.spage=e82143&rft.epage=e82143&rft.pages=e82143-e82143&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0082143&rft_dat=%3Cgale_plos_%3EA478310754%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1467678314&rft_id=info:pmid/24349203&rft_galeid=A478310754&rft_doaj_id=oai_doaj_org_article_c325b30baa1c42d6b50388db63c9c342&rfr_iscdi=true |