Peripheral denervation participates in heterotopic ossification in a spinal cord injury model
Identifieur interne : 002841 ( Pmc/Curation ); précédent : 002840; suivant : 002842Peripheral denervation participates in heterotopic ossification in a spinal cord injury model
Auteurs : Charlotte Debaud [France] ; Marjorie Salga [France] ; Laurent Begot [France] ; Xavier Holy [France] ; Malha Chedik [France] ; Nicolas De L Scalopier [France] ; Fréderic Torossian [France] ; Jean-Pierre Levesque [Australie] ; Jean-Jacques Lataillade [France] ; Marie-Caroline Le Bousse-Kerdilès [France] ; François Genêt [France]Source :
- PLoS ONE [ 1932-6203 ] ; 2017.
Abstract
We previously reported the development of a new acquired neurogenic HO (NHO) mouse model, combining spinal cord transection (SCI) and chemical muscle injury. Pathological mechanisms responsible for ectopic osteogenesis after central neurological damage are still to be elucidated. In this study, we first hypothesized that peripheral nervous system (PNS) might convey pathological signals from injured spinal cord to muscles in NHO mouse model. Secondly, we sought to determine whether SCI could lead to intramuscular modifications of BMP2 signaling pathways. Twenty one C57Bl6 mice were included in this protocol. Bilateral cardiotoxin (CTX) injection in hamstring muscles was associated with a two-stage surgical procedure, combining thoracic SCI with unilateral peripheral denervation. Volumes of HO (Bone Volume, BV) were measured 28 days after surgery using micro-computed tomography imaging techniques and histological analyses were made to confirm intramuscular osteogenesis. Volume comparisons were conducted between right and left hind limb of each animal, using a Wilcoxon signed rank test. Quantitative polymerase chain reaction (qPCR) was performed to explore intra muscular expression of BMP2, Alk3 and Id1. Nineteen mice survive the complete SCI and peripheral denervation procedure. When CTX injections were done right after surgery (n = 7), bilateral HO were detected in all animals after 28 days. Micro-CT measurements showed significantly increased BV in denervated paws (1.47 mm3 +/- 0.5) compared to contralateral sides (0.56 mm3 +/-0.4), p = 0.03. When peripheral denervation and CTX injections were performed after sham SCI surgery (n = 6), bilateral HO were present in three mice at day 28. Quantitative PCR analyses showed no changes in intra muscular BMP2 expression after SCI as compared to control mice (shamSCI). Peripheral denervation can be reliably added to spinal cord transection in NHO mouse model. This new experimental design confirms that neuro inflammatory mechanisms induced by central or peripheral nervous system injury plays a key role in triggering ectopic osteogenesis.
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DOI: 10.1371/journal.pone.0182454
PubMed: 28854256
PubMed Central: 5576715
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xml:lang="fr">France</country><wicri:regionArea>University of Versailles Saint Quentin en Yvelines, U1179 INSERM, UFR des Sciences de la Santé – Simone Veil, Montigny-le-Bretonneux</wicri:regionArea></affiliation></author><author><name sortKey="Salga, Marjorie" sort="Salga, Marjorie" uniqKey="Salga M" first="Marjorie" last="Salga">Marjorie Salga</name><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>University of Versailles Saint Quentin en Yvelines, U1179 INSERM, UFR des Sciences de la Santé – Simone Veil, Montigny-le-Bretonneux, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>University of Versailles Saint Quentin en Yvelines, U1179 INSERM, UFR des Sciences de la Santé – Simone Veil, Montigny-le-Bretonneux</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="aff003"><addr-line>Rehabilitation Service, Hôpital Raymond Poincaré, APHP, CIC-IT 1429, Garches, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Rehabilitation Service, Hôpital Raymond Poincaré, APHP, CIC-IT 1429, Garches</wicri:regionArea></affiliation></author><author><name sortKey="Begot, Laurent" sort="Begot, Laurent" uniqKey="Begot L" first="Laurent" last="Begot">Laurent Begot</name><affiliation wicri:level="1"><nlm:aff id="aff004"><addr-line>Institut de Recherche Biomédicale des Armées, Brétigny-sur-Orge, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Institut de Recherche Biomédicale des Armées, Brétigny-sur-Orge</wicri:regionArea></affiliation></author><author><name sortKey="Holy, Xavier" sort="Holy, Xavier" uniqKey="Holy X" first="Xavier" last="Holy">Xavier Holy</name><affiliation wicri:level="1"><nlm:aff id="aff004"><addr-line>Institut de Recherche Biomédicale des Armées, Brétigny-sur-Orge, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Institut de Recherche Biomédicale des Armées, Brétigny-sur-Orge</wicri:regionArea></affiliation></author><author><name sortKey="Chedik, Malha" sort="Chedik, Malha" uniqKey="Chedik M" first="Malha" last="Chedik">Malha Chedik</name><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>University of Versailles Saint Quentin en Yvelines, U1179 INSERM, UFR des Sciences de la Santé – Simone Veil, Montigny-le-Bretonneux, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>University of Versailles Saint Quentin en Yvelines, U1179 INSERM, UFR des Sciences de la Santé – Simone Veil, Montigny-le-Bretonneux</wicri:regionArea></affiliation></author><author><name sortKey="De L Scalopier, Nicolas" sort="De L Scalopier, Nicolas" uniqKey="De L Scalopier N" first="Nicolas" last="De L Scalopier">Nicolas De L Scalopier</name><affiliation wicri:level="1"><nlm:aff id="aff005"><addr-line>Orthopaedic Surgery Department, Hôpital d’Instruction des Armées PERCY, Clamart, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Orthopaedic Surgery Department, Hôpital d’Instruction des Armées PERCY, Clamart</wicri:regionArea></affiliation></author><author><name sortKey="Torossian, Frederic" sort="Torossian, Frederic" uniqKey="Torossian F" first="Fréderic" last="Torossian">Fréderic Torossian</name><affiliation wicri:level="1"><nlm:aff id="aff006"><addr-line>University of Paris-Sud, INSERM UMR-S/MD 1197, Hôpital Paul Brousse, APHP, Villejuif, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>University of Paris-Sud, INSERM UMR-S/MD 1197, Hôpital Paul Brousse, APHP, Villejuif</wicri:regionArea></affiliation></author><author><name sortKey="Levesque, Jean Pierre" sort="Levesque, Jean Pierre" uniqKey="Levesque J" first="Jean-Pierre" last="Levesque">Jean-Pierre Levesque</name><affiliation wicri:level="1"><nlm:aff id="aff007"><addr-line>Blood and Bone Diseases Program, Mater Research Institute, University of Queensland, Woolloongabba and School of Medicine, University of Queensland, Herston, Queensland, Australia</addr-line></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>Blood and Bone Diseases Program, Mater Research Institute, University of Queensland, Woolloongabba and School of Medicine, University of Queensland, Herston, Queensland</wicri:regionArea></affiliation></author><author><name sortKey="Lataillade, Jean Jacques" sort="Lataillade, Jean Jacques" uniqKey="Lataillade J" first="Jean-Jacques" last="Lataillade">Jean-Jacques Lataillade</name><affiliation wicri:level="1"><nlm:aff id="aff008"><addr-line>University of Paris-Sud, Unité mixte Inserm/SSA 1197, IRBA/CTSA/HIA Percy, École du Val de Grâce, Clamart, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>University of Paris-Sud, Unité mixte Inserm/SSA 1197, IRBA/CTSA/HIA Percy, École du Val de Grâce, Clamart</wicri:regionArea></affiliation></author><author><name sortKey="Le Bousse Kerdiles, Marie Caroline" sort="Le Bousse Kerdiles, Marie Caroline" uniqKey="Le Bousse Kerdiles M" first="Marie-Caroline" last="Le Bousse-Kerdilès">Marie-Caroline Le Bousse-Kerdilès</name><affiliation wicri:level="1"><nlm:aff id="aff006"><addr-line>University of Paris-Sud, INSERM UMR-S/MD 1197, Hôpital Paul Brousse, APHP, Villejuif, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>University of Paris-Sud, INSERM UMR-S/MD 1197, Hôpital Paul Brousse, APHP, Villejuif</wicri:regionArea></affiliation></author><author><name sortKey="Genet, Francois" sort="Genet, Francois" uniqKey="Genet F" first="François" last="Genêt">François Genêt</name><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>University of Versailles Saint Quentin en Yvelines, U1179 INSERM, UFR des Sciences de la Santé – Simone Veil, Montigny-le-Bretonneux, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>University of Versailles Saint Quentin en Yvelines, U1179 INSERM, UFR des Sciences de la Santé – Simone Veil, Montigny-le-Bretonneux</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="aff003"><addr-line>Rehabilitation Service, Hôpital Raymond Poincaré, APHP, CIC-IT 1429, Garches, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Rehabilitation Service, Hôpital Raymond Poincaré, APHP, CIC-IT 1429, Garches</wicri:regionArea></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2017">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>We previously reported the development of a new acquired neurogenic HO (NHO) mouse model, combining spinal cord transection (SCI) and chemical muscle injury. Pathological mechanisms responsible for ectopic osteogenesis after central neurological damage are still to be elucidated. In this study, we first hypothesized that peripheral nervous system (PNS) might convey pathological signals from injured spinal cord to muscles in NHO mouse model. Secondly, we sought to determine whether SCI could lead to intramuscular modifications of BMP2 signaling pathways. Twenty one C57Bl6 mice were included in this protocol. Bilateral cardiotoxin (CTX) injection in hamstring muscles was associated with a two-stage surgical procedure, combining thoracic SCI with unilateral peripheral denervation. Volumes of HO (Bone Volume, BV) were measured 28 days after surgery using micro-computed tomography imaging techniques and histological analyses were made to confirm intramuscular osteogenesis. Volume comparisons were conducted between right and left hind limb of each animal, using a Wilcoxon signed rank test. Quantitative polymerase chain reaction (qPCR) was performed to explore intra muscular expression of BMP2, Alk3 and Id1. Nineteen mice survive the complete SCI and peripheral denervation procedure. When CTX injections were done right after surgery (n = 7), bilateral HO were detected in all animals after 28 days. Micro-CT measurements showed significantly increased BV in denervated paws (1.47 mm3 +/- 0.5) compared to contralateral sides (0.56 mm3 +/-0.4), p = 0.03. When peripheral denervation and CTX injections were performed after sham SCI surgery (n = 6), bilateral HO were present in three mice at day 28. Quantitative PCR analyses showed no changes in intra muscular BMP2 expression after SCI as compared to control mice (shamSCI). Peripheral denervation can be reliably added to spinal cord transection in NHO mouse model. This new experimental design confirms that neuro inflammatory mechanisms induced by central or peripheral nervous system injury plays a key role in triggering ectopic osteogenesis.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Aubut, J Al" uniqKey="Aubut J">J-AL Aubut</name></author><author><name sortKey="Mehta, S" uniqKey="Mehta S">S Mehta</name></author><author><name sortKey="Cullen, N" uniqKey="Cullen N">N Cullen</name></author><author><name sortKey="Teasell, Rw" uniqKey="Teasell R">RW Teasell</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Haran, M" uniqKey="Haran M">M Haran</name></author><author><name sortKey="Bhuta, T" uniqKey="Bhuta T">T Bhuta</name></author><author><name sortKey="Lee, B" uniqKey="Lee B">B Lee</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shehab, D" uniqKey="Shehab D">D Shehab</name></author><author><name sortKey="Elgazzar, Ah" 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<front><journal-meta><journal-id journal-id-type="nlm-ta">PLoS One</journal-id><journal-id journal-id-type="iso-abbrev">PLoS ONE</journal-id><journal-id journal-id-type="publisher-id">plos</journal-id><journal-id journal-id-type="pmc">plosone</journal-id><journal-title-group><journal-title>PLoS ONE</journal-title></journal-title-group><issn pub-type="epub">1932-6203</issn><publisher><publisher-name>Public Library of Science</publisher-name><publisher-loc>San Francisco, CA USA</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">28854256</article-id><article-id pub-id-type="pmc">5576715</article-id><article-id pub-id-type="doi">10.1371/journal.pone.0182454</article-id><article-id pub-id-type="publisher-id">PONE-D-17-11807</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and Health Sciences</subject><subj-group><subject>Critical Care and Emergency Medicine</subject><subj-group><subject>Trauma Medicine</subject><subj-group><subject>Traumatic Injury</subject><subj-group><subject>Neurotrauma</subject><subj-group><subject>Spinal Cord Injury</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and Health Sciences</subject><subj-group><subject>Neurology</subject><subj-group><subject>Spinal Cord Injury</subject></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Research and Analysis Methods</subject><subj-group><subject>Experimental Organism Systems</subject><subj-group><subject>Model Organisms</subject><subj-group><subject>Mouse Models</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Research and Analysis Methods</subject><subj-group><subject>Model Organisms</subject><subj-group><subject>Mouse Models</subject></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Research and Analysis Methods</subject><subj-group><subject>Experimental Organism Systems</subject><subj-group><subject>Animal Models</subject><subj-group><subject>Mouse Models</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and Health Sciences</subject><subj-group><subject>Surgical and Invasive Medical Procedures</subject><subj-group><subject>Denervation</subject></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Physiology</subject><subj-group><subject>Physiological Processes</subject><subj-group><subject>Bone Remodeling</subject><subj-group><subject>Ossification</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and Health Sciences</subject><subj-group><subject>Physiology</subject><subj-group><subject>Physiological Processes</subject><subj-group><subject>Bone Remodeling</subject><subj-group><subject>Ossification</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Anatomy</subject><subj-group><subject>Nervous System</subject><subj-group><subject>Nerves</subject><subj-group><subject>Sciatic Nerves</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and Health Sciences</subject><subj-group><subject>Anatomy</subject><subj-group><subject>Nervous System</subject><subj-group><subject>Nerves</subject><subj-group><subject>Sciatic Nerves</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and Health Sciences</subject><subj-group><subject>Surgical and Invasive Medical Procedures</subject></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Anatomy</subject><subj-group><subject>Nervous System</subject><subj-group><subject>Neuroanatomy</subject><subj-group><subject>Spinal Cord</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and Health Sciences</subject><subj-group><subject>Anatomy</subject><subj-group><subject>Nervous System</subject><subj-group><subject>Neuroanatomy</subject><subj-group><subject>Spinal Cord</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Neuroscience</subject><subj-group><subject>Neuroanatomy</subject><subj-group><subject>Spinal Cord</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and life sciences</subject><subj-group><subject>Cell biology</subject><subj-group><subject>Signal transduction</subject><subj-group><subject>Cell signaling</subject><subj-group><subject>BMP signaling</subject></subj-group></subj-group></subj-group></subj-group></subj-group></article-categories><title-group><article-title>Peripheral denervation participates in heterotopic ossification in a spinal cord injury model</article-title><alt-title alt-title-type="running-head">Heterotopic ossification and nerves</alt-title></title-group><contrib-group><contrib contrib-type="author"><contrib-id authenticated="true" contrib-id-type="orcid">http://orcid.org/0000-0002-7859-8415</contrib-id><name><surname>Debaud</surname><given-names>Charlotte</given-names></name><role content-type="http://credit.casrai.org/">Conceptualization</role><role content-type="http://credit.casrai.org/">Formal analysis</role><role content-type="http://credit.casrai.org/">Investigation</role><role content-type="http://credit.casrai.org/">Methodology</role><role content-type="http://credit.casrai.org/">Project administration</role><role content-type="http://credit.casrai.org/">Resources</role><role content-type="http://credit.casrai.org/">Software</role><role content-type="http://credit.casrai.org/">Validation</role><role content-type="http://credit.casrai.org/">Visualization</role><role content-type="http://credit.casrai.org/">Writing – original draft</role><role content-type="http://credit.casrai.org/">Writing – review & editing</role><xref ref-type="aff" rid="aff001"><sup>1</sup></xref><xref ref-type="aff" rid="aff002"><sup>2</sup></xref><xref ref-type="corresp" rid="cor001">*</xref></contrib><contrib contrib-type="author"><name><surname>Salga</surname><given-names>Marjorie</given-names></name><role content-type="http://credit.casrai.org/">Formal analysis</role><role content-type="http://credit.casrai.org/">Investigation</role><role content-type="http://credit.casrai.org/">Methodology</role><xref ref-type="aff" rid="aff002"><sup>2</sup></xref><xref ref-type="aff" rid="aff003"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Begot</surname><given-names>Laurent</given-names></name><role content-type="http://credit.casrai.org/">Formal analysis</role><role content-type="http://credit.casrai.org/">Investigation</role><role content-type="http://credit.casrai.org/">Methodology</role><role content-type="http://credit.casrai.org/">Resources</role><role content-type="http://credit.casrai.org/">Software</role><xref ref-type="aff" rid="aff004"><sup>4</sup></xref></contrib><contrib contrib-type="author"><name><surname>Holy</surname><given-names>Xavier</given-names></name><role content-type="http://credit.casrai.org/">Formal analysis</role><role content-type="http://credit.casrai.org/">Methodology</role><role content-type="http://credit.casrai.org/">Resources</role><xref ref-type="aff" rid="aff004"><sup>4</sup></xref></contrib><contrib contrib-type="author"><name><surname>Chedik</surname><given-names>Malha</given-names></name><role content-type="http://credit.casrai.org/">Resources</role><xref ref-type="aff" rid="aff002"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>de l’Escalopier</surname><given-names>Nicolas</given-names></name><role content-type="http://credit.casrai.org/">Resources</role><xref ref-type="aff" rid="aff005"><sup>5</sup></xref></contrib><contrib contrib-type="author"><name><surname>Torossian</surname><given-names>Fréderic</given-names></name><role content-type="http://credit.casrai.org/">Formal analysis</role><role content-type="http://credit.casrai.org/">Methodology</role><role content-type="http://credit.casrai.org/">Resources</role><role content-type="http://credit.casrai.org/">Software</role><role content-type="http://credit.casrai.org/">Visualization</role><role content-type="http://credit.casrai.org/">Writing – review & editing</role><xref ref-type="aff" rid="aff006"><sup>6</sup></xref></contrib><contrib contrib-type="author"><name><surname>Levesque</surname><given-names>Jean-Pierre</given-names></name><role content-type="http://credit.casrai.org/">Funding acquisition</role><role content-type="http://credit.casrai.org/">Methodology</role><role content-type="http://credit.casrai.org/">Supervision</role><xref ref-type="aff" rid="aff007"><sup>7</sup></xref></contrib><contrib contrib-type="author"><name><surname>Lataillade</surname><given-names>Jean-Jacques</given-names></name><role content-type="http://credit.casrai.org/">Funding acquisition</role><role content-type="http://credit.casrai.org/">Methodology</role><role content-type="http://credit.casrai.org/">Supervision</role><xref ref-type="aff" rid="aff008"><sup>8</sup></xref></contrib><contrib contrib-type="author"><name><surname>Le Bousse-Kerdilès</surname><given-names>Marie-Caroline</given-names></name><role content-type="http://credit.casrai.org/">Formal analysis</role><role content-type="http://credit.casrai.org/">Funding acquisition</role><role content-type="http://credit.casrai.org/">Methodology</role><role content-type="http://credit.casrai.org/">Resources</role><role content-type="http://credit.casrai.org/">Supervision</role><role content-type="http://credit.casrai.org/">Validation</role><role content-type="http://credit.casrai.org/">Visualization</role><role content-type="http://credit.casrai.org/">Writing – review & editing</role><xref ref-type="aff" rid="aff006"><sup>6</sup></xref></contrib><contrib contrib-type="author"><name><surname>Genêt</surname><given-names>François</given-names></name><role content-type="http://credit.casrai.org/">Conceptualization</role><role content-type="http://credit.casrai.org/">Funding acquisition</role><role content-type="http://credit.casrai.org/">Methodology</role><role content-type="http://credit.casrai.org/">Project administration</role><role content-type="http://credit.casrai.org/">Resources</role><role content-type="http://credit.casrai.org/">Supervision</role><role content-type="http://credit.casrai.org/">Validation</role><role content-type="http://credit.casrai.org/">Visualization</role><role content-type="http://credit.casrai.org/">Writing – original draft</role><role content-type="http://credit.casrai.org/">Writing – review & editing</role><xref ref-type="aff" rid="aff002"><sup>2</sup></xref><xref ref-type="aff" rid="aff003"><sup>3</sup></xref></contrib></contrib-group><aff id="aff001"><label>1</label><addr-line>Spine Division Orthopaedic Surgery Department, Hôpital Européen Georges Pompidou, APHP, Paris, France</addr-line></aff><aff id="aff002"><label>2</label><addr-line>University of Versailles Saint Quentin en Yvelines, U1179 INSERM, UFR des Sciences de la Santé – Simone Veil, Montigny-le-Bretonneux, France</addr-line></aff><aff id="aff003"><label>3</label><addr-line>Rehabilitation Service, Hôpital Raymond Poincaré, APHP, CIC-IT 1429, Garches, France</addr-line></aff><aff id="aff004"><label>4</label><addr-line>Institut de Recherche Biomédicale des Armées, Brétigny-sur-Orge, France</addr-line></aff><aff id="aff005"><label>5</label><addr-line>Orthopaedic Surgery Department, Hôpital d’Instruction des Armées PERCY, Clamart, France</addr-line></aff><aff id="aff006"><label>6</label><addr-line>University of Paris-Sud, INSERM UMR-S/MD 1197, Hôpital Paul Brousse, APHP, Villejuif, France</addr-line></aff><aff id="aff007"><label>7</label><addr-line>Blood and Bone Diseases Program, Mater Research Institute, University of Queensland, Woolloongabba and School of Medicine, University of Queensland, Herston, Queensland, Australia</addr-line></aff><aff id="aff008"><label>8</label><addr-line>University of Paris-Sud, Unité mixte Inserm/SSA 1197, IRBA/CTSA/HIA Percy, École du Val de Grâce, Clamart, France</addr-line></aff><contrib-group><contrib contrib-type="editor"><name><surname>Asakura</surname><given-names>Atsushi</given-names></name><role>Editor</role><xref ref-type="aff" rid="edit1"></xref></contrib></contrib-group><aff id="edit1"><addr-line>University of Minnesota Medical Center, UNITED STATES</addr-line></aff><author-notes><fn fn-type="COI-statement" id="coi001"><p><bold>Competing Interests: </bold>The authors have declared that no competing interests exist.</p></fn><corresp id="cor001">* E-mail: <email>charlotte.debaud@aphp.fr</email></corresp></author-notes><pub-date pub-type="epub"><day>30</day><month>8</month><year>2017</year></pub-date><pub-date pub-type="collection"><year>2017</year></pub-date><volume>12</volume><issue>8</issue><elocation-id>e0182454</elocation-id><history><date date-type="received"><day>28</day><month>3</month><year>2017</year></date><date date-type="accepted"><day>18</day><month>7</month><year>2017</year></date></history><permissions><copyright-statement>© 2017 Debaud et al</copyright-statement><copyright-year>2017</copyright-year><copyright-holder>Debaud et al</copyright-holder><license xlink:href="http://creativecommons.org/licenses/by/4.0/"><license-p>This is an open access article distributed under the terms of the <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License</ext-link>, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</license-p></license></permissions><self-uri content-type="pdf" xlink:href="pone.0182454.pdf"></self-uri><abstract><p>We previously reported the development of a new acquired neurogenic HO (NHO) mouse model, combining spinal cord transection (SCI) and chemical muscle injury. Pathological mechanisms responsible for ectopic osteogenesis after central neurological damage are still to be elucidated. In this study, we first hypothesized that peripheral nervous system (PNS) might convey pathological signals from injured spinal cord to muscles in NHO mouse model. Secondly, we sought to determine whether SCI could lead to intramuscular modifications of BMP2 signaling pathways. Twenty one C57Bl6 mice were included in this protocol. Bilateral cardiotoxin (CTX) injection in hamstring muscles was associated with a two-stage surgical procedure, combining thoracic SCI with unilateral peripheral denervation. Volumes of HO (Bone Volume, BV) were measured 28 days after surgery using micro-computed tomography imaging techniques and histological analyses were made to confirm intramuscular osteogenesis. Volume comparisons were conducted between right and left hind limb of each animal, using a Wilcoxon signed rank test. Quantitative polymerase chain reaction (qPCR) was performed to explore intra muscular expression of BMP2, Alk3 and Id1. Nineteen mice survive the complete SCI and peripheral denervation procedure. When CTX injections were done right after surgery (n = 7), bilateral HO were detected in all animals after 28 days. Micro-CT measurements showed significantly increased BV in denervated paws (1.47 mm3 +/- 0.5) compared to contralateral sides (0.56 mm3 +/-0.4), p = 0.03. When peripheral denervation and CTX injections were performed after sham SCI surgery (n = 6), bilateral HO were present in three mice at day 28. Quantitative PCR analyses showed no changes in intra muscular BMP2 expression after SCI as compared to control mice (shamSCI). Peripheral denervation can be reliably added to spinal cord transection in NHO mouse model. This new experimental design confirms that neuro inflammatory mechanisms induced by central or peripheral nervous system injury plays a key role in triggering ectopic osteogenesis.</p></abstract><funding-group><funding-statement>The authors received no specific funding for this work.</funding-statement></funding-group><counts><fig-count count="5"></fig-count><table-count count="1"></table-count><page-count count="14"></page-count></counts><custom-meta-group><custom-meta id="data-availability"><meta-name>Data Availability</meta-name><meta-value>All relevant data are within the paper and its Supporting Information files.</meta-value></custom-meta></custom-meta-group></article-meta><notes><title>Data Availability</title><p>All relevant data are within the paper and its Supporting Information files.</p></notes></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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