Contribution of fibronectin and vitronectin to the adhesion and morphology of MC3T3-E1 osteoblastic cells to poly(NaSS) grafted Ti6Al4V.
Identifieur interne : 002549 ( PubMed/Curation ); précédent : 002548; suivant : 002550Contribution of fibronectin and vitronectin to the adhesion and morphology of MC3T3-E1 osteoblastic cells to poly(NaSS) grafted Ti6Al4V.
Auteurs : Helena P. Felgueiras [France] ; Margaret D M. Evans [Australie] ; Véronique Migonney [France]Source :
- Acta biomaterialia [ 1878-7568 ] ; 2015.
Descripteurs français
- KwdFr :
- MESH :
- cytologie : Ostéoblastes.
- métabolisme : Fibronectines, Vitronectine.
- physiologie : Intégrines.
- Adhérence cellulaire, Animaux, Cellules 3T3, Propriétés de surface, Souris, Titane.
English descriptors
- KwdEn :
- MESH :
- chemical , metabolism : Fibronectins, Vitronectin.
- chemical , physiology : Integrins.
- cytology : Osteoblasts.
- 3T3 Cells, Animals, Cell Adhesion, Mice, Surface Properties, Titanium.
Abstract
This study is focused on understanding the underlying mechanisms involved in the improved in vitro and in vivo responses of osteoblasts on poly(sodium styrene sulfonate) (poly(NaSS)) functionalized Ti6Al4V surfaces. We probed the contribution of cell-adhesive glycoproteins fibronectin (Fn) and vitronectin (Vn) in the initial adhesion of MC3T3-E1 osteoblastic cells to poly(NaSS) functionalized and control Ti6Al4V surfaces. Firstly, culture media containing serum depleted of Fn and Vn (DD) were used to establish the contribution of Fn and Vn in the adhesion and spreading of cells on poly(NaSS) grafted and control surfaces. Compared to ungrafted surfaces, poly(NaSS) grafted surfaces enhanced the levels of cell adhesion, cell spreading and the formation of intracellular actin cytoskeleton and focal contacts in serum treatments where Fn or Vn were present (FBS, DD+Fn, DD+Vn). Cell responses to Fn were more significant than to Vn. Secondly, blocking Fn and Vn integrin receptors using antibodies to α5β1 (Fn) and αvβ1 (Vn) showed that adhesion of cells to poly(NaSS) grafted surfaces principally involved the Fn integrin receptor α5β1. Thirdly, blocking of the heparin and cell-binding regions of Fn molecule (RGD, C-HB, N-HB) showed that grafting with poly(NaSS) altered the conformation of Fn. Together these outcomes explained why the presence of sulfonate (SO3(-)) groups grafted on the Ti6Al4V surface enhanced the early cell adhesion and spreading processes which determine clinical success for applications that require osseointegration.
DOI: 10.1016/j.actbio.2015.09.030
PubMed: 26415777
Links toward previous steps (curation, corpus...)
- to stream PubMed, to step Corpus: Pour aller vers cette notice dans l'étape Curation :002618
Links to Exploration step
pubmed:26415777Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Contribution of fibronectin and vitronectin to the adhesion and morphology of MC3T3-E1 osteoblastic cells to poly(NaSS) grafted Ti6Al4V.</title>
<author><name sortKey="Felgueiras, Helena P" sort="Felgueiras, Helena P" uniqKey="Felgueiras H" first="Helena P" last="Felgueiras">Helena P. Felgueiras</name>
<affiliation wicri:level="1"><nlm:affiliation>Laboratory of Biomaterials and Specialty Polymers, LBPS-CSPBAT CNRS UMR 7244, Institut Galilée, Université Paris 13, Sorbonne Paris Cité, 93430 Villetaneuse, France.</nlm:affiliation>
<country xml:lang="fr">France</country>
<wicri:regionArea>Laboratory of Biomaterials and Specialty Polymers, LBPS-CSPBAT CNRS UMR 7244, Institut Galilée, Université Paris 13, Sorbonne Paris Cité, 93430 Villetaneuse</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Evans, Margaret D M" sort="Evans, Margaret D M" uniqKey="Evans M" first="Margaret D M" last="Evans">Margaret D M. Evans</name>
<affiliation wicri:level="1"><nlm:affiliation>CSIRO Biomedical Materials Manufacturing Program, 11 Julius Avenue, North Ryde, Sydney, NSW 2113, Australia.</nlm:affiliation>
<country xml:lang="fr">Australie</country>
<wicri:regionArea>CSIRO Biomedical Materials Manufacturing Program, 11 Julius Avenue, North Ryde, Sydney, NSW 2113</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Migonney, Veronique" sort="Migonney, Veronique" uniqKey="Migonney V" first="Véronique" last="Migonney">Véronique Migonney</name>
<affiliation wicri:level="1"><nlm:affiliation>Laboratory of Biomaterials and Specialty Polymers, LBPS-CSPBAT CNRS UMR 7244, Institut Galilée, Université Paris 13, Sorbonne Paris Cité, 93430 Villetaneuse, France. Electronic address: veronique.migonney@univ-paris13.fr.</nlm:affiliation>
<country xml:lang="fr">France</country>
<wicri:regionArea>Laboratory of Biomaterials and Specialty Polymers, LBPS-CSPBAT CNRS UMR 7244, Institut Galilée, Université Paris 13, Sorbonne Paris Cité, 93430 Villetaneuse</wicri:regionArea>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PubMed</idno>
<date when="2015">2015</date>
<idno type="RBID">pubmed:26415777</idno>
<idno type="pmid">26415777</idno>
<idno type="doi">10.1016/j.actbio.2015.09.030</idno>
<idno type="wicri:Area/PubMed/Corpus">002618</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002618</idno>
<idno type="wicri:Area/PubMed/Curation">002549</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002549</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en">Contribution of fibronectin and vitronectin to the adhesion and morphology of MC3T3-E1 osteoblastic cells to poly(NaSS) grafted Ti6Al4V.</title>
<author><name sortKey="Felgueiras, Helena P" sort="Felgueiras, Helena P" uniqKey="Felgueiras H" first="Helena P" last="Felgueiras">Helena P. Felgueiras</name>
<affiliation wicri:level="1"><nlm:affiliation>Laboratory of Biomaterials and Specialty Polymers, LBPS-CSPBAT CNRS UMR 7244, Institut Galilée, Université Paris 13, Sorbonne Paris Cité, 93430 Villetaneuse, France.</nlm:affiliation>
<country xml:lang="fr">France</country>
<wicri:regionArea>Laboratory of Biomaterials and Specialty Polymers, LBPS-CSPBAT CNRS UMR 7244, Institut Galilée, Université Paris 13, Sorbonne Paris Cité, 93430 Villetaneuse</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Evans, Margaret D M" sort="Evans, Margaret D M" uniqKey="Evans M" first="Margaret D M" last="Evans">Margaret D M. Evans</name>
<affiliation wicri:level="1"><nlm:affiliation>CSIRO Biomedical Materials Manufacturing Program, 11 Julius Avenue, North Ryde, Sydney, NSW 2113, Australia.</nlm:affiliation>
<country xml:lang="fr">Australie</country>
<wicri:regionArea>CSIRO Biomedical Materials Manufacturing Program, 11 Julius Avenue, North Ryde, Sydney, NSW 2113</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Migonney, Veronique" sort="Migonney, Veronique" uniqKey="Migonney V" first="Véronique" last="Migonney">Véronique Migonney</name>
<affiliation wicri:level="1"><nlm:affiliation>Laboratory of Biomaterials and Specialty Polymers, LBPS-CSPBAT CNRS UMR 7244, Institut Galilée, Université Paris 13, Sorbonne Paris Cité, 93430 Villetaneuse, France. Electronic address: veronique.migonney@univ-paris13.fr.</nlm:affiliation>
<country xml:lang="fr">France</country>
<wicri:regionArea>Laboratory of Biomaterials and Specialty Polymers, LBPS-CSPBAT CNRS UMR 7244, Institut Galilée, Université Paris 13, Sorbonne Paris Cité, 93430 Villetaneuse</wicri:regionArea>
</affiliation>
</author>
</analytic>
<series><title level="j">Acta biomaterialia</title>
<idno type="eISSN">1878-7568</idno>
<imprint><date when="2015" type="published">2015</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>3T3 Cells</term>
<term>Animals</term>
<term>Cell Adhesion</term>
<term>Fibronectins (metabolism)</term>
<term>Integrins (physiology)</term>
<term>Mice</term>
<term>Osteoblasts (cytology)</term>
<term>Surface Properties</term>
<term>Titanium</term>
<term>Vitronectin (metabolism)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Adhérence cellulaire</term>
<term>Animaux</term>
<term>Cellules 3T3</term>
<term>Fibronectines (métabolisme)</term>
<term>Intégrines (physiologie)</term>
<term>Ostéoblastes (cytologie)</term>
<term>Propriétés de surface</term>
<term>Souris</term>
<term>Titane</term>
<term>Vitronectine (métabolisme)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Fibronectins</term>
<term>Vitronectin</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="physiology" xml:lang="en"><term>Integrins</term>
</keywords>
<keywords scheme="MESH" qualifier="cytologie" xml:lang="fr"><term>Ostéoblastes</term>
</keywords>
<keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Osteoblasts</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Fibronectines</term>
<term>Vitronectine</term>
</keywords>
<keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Intégrines</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>3T3 Cells</term>
<term>Animals</term>
<term>Cell Adhesion</term>
<term>Mice</term>
<term>Surface Properties</term>
<term>Titanium</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Adhérence cellulaire</term>
<term>Animaux</term>
<term>Cellules 3T3</term>
<term>Propriétés de surface</term>
<term>Souris</term>
<term>Titane</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">This study is focused on understanding the underlying mechanisms involved in the improved in vitro and in vivo responses of osteoblasts on poly(sodium styrene sulfonate) (poly(NaSS)) functionalized Ti6Al4V surfaces. We probed the contribution of cell-adhesive glycoproteins fibronectin (Fn) and vitronectin (Vn) in the initial adhesion of MC3T3-E1 osteoblastic cells to poly(NaSS) functionalized and control Ti6Al4V surfaces. Firstly, culture media containing serum depleted of Fn and Vn (DD) were used to establish the contribution of Fn and Vn in the adhesion and spreading of cells on poly(NaSS) grafted and control surfaces. Compared to ungrafted surfaces, poly(NaSS) grafted surfaces enhanced the levels of cell adhesion, cell spreading and the formation of intracellular actin cytoskeleton and focal contacts in serum treatments where Fn or Vn were present (FBS, DD+Fn, DD+Vn). Cell responses to Fn were more significant than to Vn. Secondly, blocking Fn and Vn integrin receptors using antibodies to α5β1 (Fn) and αvβ1 (Vn) showed that adhesion of cells to poly(NaSS) grafted surfaces principally involved the Fn integrin receptor α5β1. Thirdly, blocking of the heparin and cell-binding regions of Fn molecule (RGD, C-HB, N-HB) showed that grafting with poly(NaSS) altered the conformation of Fn. Together these outcomes explained why the presence of sulfonate (SO3(-)) groups grafted on the Ti6Al4V surface enhanced the early cell adhesion and spreading processes which determine clinical success for applications that require osseointegration.</div>
</front>
</TEI>
<pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">26415777</PMID>
<DateCreated><Year>2015</Year>
<Month>11</Month>
<Day>16</Day>
</DateCreated>
<DateCompleted><Year>2016</Year>
<Month>09</Month>
<Day>23</Day>
</DateCompleted>
<DateRevised><Year>2017</Year>
<Month>08</Month>
<Day>13</Day>
</DateRevised>
<Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1878-7568</ISSN>
<JournalIssue CitedMedium="Internet"><Volume>28</Volume>
<PubDate><Year>2015</Year>
<Month>Dec</Month>
</PubDate>
</JournalIssue>
<Title>Acta biomaterialia</Title>
<ISOAbbreviation>Acta Biomater</ISOAbbreviation>
</Journal>
<ArticleTitle>Contribution of fibronectin and vitronectin to the adhesion and morphology of MC3T3-E1 osteoblastic cells to poly(NaSS) grafted Ti6Al4V.</ArticleTitle>
<Pagination><MedlinePgn>225-233</MedlinePgn>
</Pagination>
<ELocationID EIdType="pii" ValidYN="Y">S1742-7061(15)30123-9</ELocationID>
<ELocationID EIdType="doi" ValidYN="Y">10.1016/j.actbio.2015.09.030</ELocationID>
<Abstract><AbstractText Label="UNLABELLED">This study is focused on understanding the underlying mechanisms involved in the improved in vitro and in vivo responses of osteoblasts on poly(sodium styrene sulfonate) (poly(NaSS)) functionalized Ti6Al4V surfaces. We probed the contribution of cell-adhesive glycoproteins fibronectin (Fn) and vitronectin (Vn) in the initial adhesion of MC3T3-E1 osteoblastic cells to poly(NaSS) functionalized and control Ti6Al4V surfaces. Firstly, culture media containing serum depleted of Fn and Vn (DD) were used to establish the contribution of Fn and Vn in the adhesion and spreading of cells on poly(NaSS) grafted and control surfaces. Compared to ungrafted surfaces, poly(NaSS) grafted surfaces enhanced the levels of cell adhesion, cell spreading and the formation of intracellular actin cytoskeleton and focal contacts in serum treatments where Fn or Vn were present (FBS, DD+Fn, DD+Vn). Cell responses to Fn were more significant than to Vn. Secondly, blocking Fn and Vn integrin receptors using antibodies to α5β1 (Fn) and αvβ1 (Vn) showed that adhesion of cells to poly(NaSS) grafted surfaces principally involved the Fn integrin receptor α5β1. Thirdly, blocking of the heparin and cell-binding regions of Fn molecule (RGD, C-HB, N-HB) showed that grafting with poly(NaSS) altered the conformation of Fn. Together these outcomes explained why the presence of sulfonate (SO3(-)) groups grafted on the Ti6Al4V surface enhanced the early cell adhesion and spreading processes which determine clinical success for applications that require osseointegration.</AbstractText>
<AbstractText Label="STATEMENT OF SIGNIFICANCE" NlmCategory="UNASSIGNED">This study is devoted to the basic analysis of the mechanism at the origin of the improved in vitro and in vivo osteoblast cell responses exhibited by poly(sodium styrene sulfonate) (poly(NaSS)) functionalized Ti6Al4V surfaces. The aim was to probe the contribution of cell adhesive glycoproteins fibronectin and vitronectin in the initial adhesion of MC3T3-E1 osteoblastic cells to poly(NaSS) functionalized Ti6Al4V surfaces. The outcomes of this research explained why the presence of SO3(-) (sulfonate) groups grafted on the Ti6Al4V surface enhanced the early cell adhesion and spreading processes which determine clinical success for applications that require osseointegration. This work is a step further in the research of poly(NaSS), a very promising bioactive polymer with potential to the orthopedic and dental fields.</AbstractText>
<CopyrightInformation>Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.</CopyrightInformation>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Felgueiras</LastName>
<ForeName>Helena P</ForeName>
<Initials>HP</Initials>
<AffiliationInfo><Affiliation>Laboratory of Biomaterials and Specialty Polymers, LBPS-CSPBAT CNRS UMR 7244, Institut Galilée, Université Paris 13, Sorbonne Paris Cité, 93430 Villetaneuse, France.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Evans</LastName>
<ForeName>Margaret D M</ForeName>
<Initials>MDM</Initials>
<AffiliationInfo><Affiliation>CSIRO Biomedical Materials Manufacturing Program, 11 Julius Avenue, North Ryde, Sydney, NSW 2113, Australia.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Migonney</LastName>
<ForeName>Véronique</ForeName>
<Initials>V</Initials>
<AffiliationInfo><Affiliation>Laboratory of Biomaterials and Specialty Polymers, LBPS-CSPBAT CNRS UMR 7244, Institut Galilée, Université Paris 13, Sorbonne Paris Cité, 93430 Villetaneuse, France. Electronic address: veronique.migonney@univ-paris13.fr.</Affiliation>
</AffiliationInfo>
</Author>
</AuthorList>
<Language>eng</Language>
<PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType>
<PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType>
</PublicationTypeList>
<ArticleDate DateType="Electronic"><Year>2015</Year>
<Month>09</Month>
<Day>28</Day>
</ArticleDate>
</Article>
<MedlineJournalInfo><Country>England</Country>
<MedlineTA>Acta Biomater</MedlineTA>
<NlmUniqueID>101233144</NlmUniqueID>
<ISSNLinking>1742-7061</ISSNLinking>
</MedlineJournalInfo>
<ChemicalList><Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D005353">Fibronectins</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D016023">Integrins</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D019096">Vitronectin</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>12743-70-3</RegistryNumber>
<NameOfSubstance UI="C031462">titanium alloy (TiAl6V4)</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>D1JT611TNE</RegistryNumber>
<NameOfSubstance UI="D014025">Titanium</NameOfSubstance>
</Chemical>
</ChemicalList>
<CitationSubset>IM</CitationSubset>
<MeshHeadingList><MeshHeading><DescriptorName UI="D016475" MajorTopicYN="N">3T3 Cells</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D002448" MajorTopicYN="Y">Cell Adhesion</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D005353" MajorTopicYN="N">Fibronectins</DescriptorName>
<QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D016023" MajorTopicYN="N">Integrins</DescriptorName>
<QualifierName UI="Q000502" MajorTopicYN="N">physiology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D051379" MajorTopicYN="N">Mice</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D010006" MajorTopicYN="N">Osteoblasts</DescriptorName>
<QualifierName UI="Q000166" MajorTopicYN="Y">cytology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D013499" MajorTopicYN="N">Surface Properties</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D014025" MajorTopicYN="Y">Titanium</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D019096" MajorTopicYN="N">Vitronectin</DescriptorName>
<QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName>
</MeshHeading>
</MeshHeadingList>
<KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Cell adhesion</Keyword>
<Keyword MajorTopicYN="N">Depleted serum</Keyword>
<Keyword MajorTopicYN="N">Fibronectin</Keyword>
<Keyword MajorTopicYN="N">Osteoblast</Keyword>
<Keyword MajorTopicYN="N">Poly(NaSS)</Keyword>
<Keyword MajorTopicYN="N">Ti6Al4V</Keyword>
<Keyword MajorTopicYN="N">Vitronectin</Keyword>
</KeywordList>
</MedlineCitation>
<PubmedData><History><PubMedPubDate PubStatus="received"><Year>2015</Year>
<Month>06</Month>
<Day>01</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="revised"><Year>2015</Year>
<Month>08</Month>
<Day>27</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="accepted"><Year>2015</Year>
<Month>09</Month>
<Day>25</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="entrez"><Year>2015</Year>
<Month>9</Month>
<Day>30</Day>
<Hour>6</Hour>
<Minute>0</Minute>
</PubMedPubDate>
<PubMedPubDate PubStatus="pubmed"><Year>2015</Year>
<Month>9</Month>
<Day>30</Day>
<Hour>6</Hour>
<Minute>0</Minute>
</PubMedPubDate>
<PubMedPubDate PubStatus="medline"><Year>2016</Year>
<Month>9</Month>
<Day>24</Day>
<Hour>6</Hour>
<Minute>0</Minute>
</PubMedPubDate>
</History>
<PublicationStatus>ppublish</PublicationStatus>
<ArticleIdList><ArticleId IdType="pubmed">26415777</ArticleId>
<ArticleId IdType="pii">S1742-7061(15)30123-9</ArticleId>
<ArticleId IdType="doi">10.1016/j.actbio.2015.09.030</ArticleId>
</ArticleIdList>
</PubmedData>
</pubmed>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Asie/explor/AustralieFrV1/Data/PubMed/Curation
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 002549 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/PubMed/Curation/biblio.hfd -nk 002549 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Wicri/Asie |area= AustralieFrV1 |flux= PubMed |étape= Curation |type= RBID |clé= pubmed:26415777 |texte= Contribution of fibronectin and vitronectin to the adhesion and morphology of MC3T3-E1 osteoblastic cells to poly(NaSS) grafted Ti6Al4V. }}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/PubMed/Curation/RBID.i -Sk "pubmed:26415777" \ | HfdSelect -Kh $EXPLOR_AREA/Data/PubMed/Curation/biblio.hfd \ | NlmPubMed2Wicri -a AustralieFrV1
![]() | This area was generated with Dilib version V0.6.33. | ![]() |