Immobilization of chitosan film containing semaphorin 3A onto a microarc oxidized titanium implant surface via silane reaction to improve MG63 osteogenic differentiation
Identifieur interne : 001420 ( Pmc/Checkpoint ); précédent : 001419; suivant : 001421Immobilization of chitosan film containing semaphorin 3A onto a microarc oxidized titanium implant surface via silane reaction to improve MG63 osteogenic differentiation
Auteurs : Kaixiu Fang [République populaire de Chine] ; Wen Song [République populaire de Chine] ; Lifeng Wang [République populaire de Chine] ; Sen Jia [République populaire de Chine] ; Hongbo Wei [République populaire de Chine] ; Shuai Ren [République populaire de Chine] ; Xiaoru Xu [République populaire de Chine] ; Yingliang Song [République populaire de Chine]Source :
- International Journal of Nanomedicine [ 1176-9114 ] ; 2014.
Abstract
Improving osseointegration of extensively used titanium (Ti) implants still remains a main theme in implantology. Recently, grafting biomolecules onto a Ti surface has attracted more attention due to their direct participation in the osseointegration process around the implant. Semaphorin 3A (Sema3A) is a new proven osteoprotection molecule and is considered to be a promising therapeutic agent in bone diseases, but how to immobilize the protein onto a Ti surface to acquire a long-term effect is poorly defined. In our study, we tried to use chitosan to wrap Sema3A (CS/Sema) and connect to the microarc oxidized Ti surface via silane glutaraldehyde coupling. The microarc oxidization could formulate porous topography on a Ti surface, and the covalently bonded coating was homogeneously covered on the ridges between the pores without significant influence on the original topography. A burst release of Sema3A was observed in the first few days in phosphate-buffered saline and could be maintained for >2 weeks. Coating in phosphate-buffered saline containing lysozyme was similar, but the release rate was much more rapid. The coating did not significantly affect cellular adhesion, viability, or cytoskeleton arrangement, but the osteogenic-related gene expression was dramatically increased and calcium deposition was also abundantly detected. In conclusion, covalent bonding of CS/Sema could strongly improve osteogenic differentiation of osteoblasts and might be applied for Ti implant surface biofunctionalization.
Url:
DOI: 10.2147/IJN.S68895
PubMed: 25336945
PubMed Central: 4200022
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Song</name><affiliation wicri:level="1"><nlm:aff id="af2-ijn-9-4649">State Key Laboratory of Military Stomatology, Department of Prosthetic Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Military Stomatology, Department of Prosthetic Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an</wicri:regionArea><wicri:noRegion>Xi’an</wicri:noRegion></affiliation></author><author><name sortKey="Wang, Lifeng" sort="Wang, Lifeng" uniqKey="Wang L" first="Lifeng" last="Wang">Lifeng Wang</name><affiliation wicri:level="1"><nlm:aff id="af1-ijn-9-4649">State Key Laboratory of Military Stomatology, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</nlm:aff><country xml:lang="fr">République populaire de 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Chine</country><wicri:regionArea>State Key Laboratory of Military Stomatology, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an</wicri:regionArea><wicri:noRegion>Xi’an</wicri:noRegion></affiliation></author><author><name sortKey="Song, Wen" sort="Song, Wen" uniqKey="Song W" first="Wen" last="Song">Wen Song</name><affiliation wicri:level="1"><nlm:aff id="af2-ijn-9-4649">State Key Laboratory of Military Stomatology, Department of Prosthetic Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Military Stomatology, Department of Prosthetic Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an</wicri:regionArea><wicri:noRegion>Xi’an</wicri:noRegion></affiliation></author><author><name sortKey="Wang, Lifeng" sort="Wang, Lifeng" uniqKey="Wang L" first="Lifeng" last="Wang">Lifeng Wang</name><affiliation wicri:level="1"><nlm:aff id="af1-ijn-9-4649">State Key Laboratory of Military Stomatology, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Military Stomatology, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an</wicri:regionArea><wicri:noRegion>Xi’an</wicri:noRegion></affiliation></author><author><name sortKey="Jia, Sen" sort="Jia, Sen" uniqKey="Jia S" first="Sen" last="Jia">Sen Jia</name><affiliation wicri:level="1"><nlm:aff id="af3-ijn-9-4649">State Key Laboratory of Military Stomatology, Department of Oral and Maxillofacial Surgery, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Military Stomatology, Department of Oral and Maxillofacial Surgery, School of Stomatology, Fourth Military Medical University, Xi’an</wicri:regionArea><wicri:noRegion>Xi’an</wicri:noRegion></affiliation></author><author><name sortKey="Wei, Hongbo" sort="Wei, Hongbo" uniqKey="Wei H" first="Hongbo" last="Wei">Hongbo Wei</name><affiliation wicri:level="1"><nlm:aff id="af1-ijn-9-4649">State Key Laboratory of Military Stomatology, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Military Stomatology, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an</wicri:regionArea><wicri:noRegion>Xi’an</wicri:noRegion></affiliation></author><author><name sortKey="Ren, Shuai" sort="Ren, Shuai" uniqKey="Ren S" first="Shuai" last="Ren">Shuai Ren</name><affiliation wicri:level="1"><nlm:aff id="af1-ijn-9-4649">State Key Laboratory of Military Stomatology, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Military Stomatology, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an</wicri:regionArea><wicri:noRegion>Xi’an</wicri:noRegion></affiliation></author><author><name sortKey="Xu, Xiaoru" sort="Xu, Xiaoru" uniqKey="Xu X" first="Xiaoru" last="Xu">Xiaoru Xu</name><affiliation wicri:level="1"><nlm:aff id="af1-ijn-9-4649">State Key Laboratory of Military Stomatology, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Military Stomatology, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an</wicri:regionArea><wicri:noRegion>Xi’an</wicri:noRegion></affiliation></author><author><name sortKey="Song, Yingliang" sort="Song, Yingliang" uniqKey="Song Y" first="Yingliang" last="Song">Yingliang Song</name><affiliation wicri:level="1"><nlm:aff id="af1-ijn-9-4649">State Key Laboratory of Military Stomatology, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Military Stomatology, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an</wicri:regionArea><wicri:noRegion>Xi’an</wicri:noRegion></affiliation></author></analytic><series><title level="j">International Journal of Nanomedicine</title><idno type="ISSN">1176-9114</idno><idno type="eISSN">1178-2013</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Improving osseointegration of extensively used titanium (Ti) implants still remains a main theme in implantology. Recently, grafting biomolecules onto a Ti surface has attracted more attention due to their direct participation in the osseointegration process around the implant. Semaphorin 3A (Sema3A) is a new proven osteoprotection molecule and is considered to be a promising therapeutic agent in bone diseases, but how to immobilize the protein onto a Ti surface to acquire a long-term effect is poorly defined. In our study, we tried to use chitosan to wrap Sema3A (CS/Sema) and connect to the microarc oxidized Ti surface via silane glutaraldehyde coupling. The microarc oxidization could formulate porous topography on a Ti surface, and the covalently bonded coating was homogeneously covered on the ridges between the pores without significant influence on the original topography. A burst release of Sema3A was observed in the first few days in phosphate-buffered saline and could be maintained for >2 weeks. Coating in phosphate-buffered saline containing lysozyme was similar, but the release rate was much more rapid. The coating did not significantly affect cellular adhesion, viability, or cytoskeleton arrangement, but the osteogenic-related gene expression was dramatically increased and calcium deposition was also abundantly detected. In conclusion, covalent bonding of CS/Sema could strongly improve osteogenic differentiation of osteoblasts and might be applied for Ti implant surface biofunctionalization.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Elias, Cn" uniqKey="Elias C">CN Elias</name></author><author><name sortKey="Lima, Jhc" uniqKey="Lima J">JHC Lima</name></author><author><name sortKey="Valiev, R" uniqKey="Valiev R">R Valiev</name></author><author><name sortKey="Meyers, Ma" uniqKey="Meyers M">MA Meyers</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Buser, D" uniqKey="Buser D">D Buser</name></author><author><name sortKey="Janner, Sf" uniqKey="Janner S">SF Janner</name></author><author><name sortKey="Wittneben, Jg" uniqKey="Wittneben J">JG Wittneben</name></author><author><name sortKey="Bragger, U" uniqKey="Bragger U">U Bragger</name></author><author><name sortKey="Ramseier, Ca" uniqKey="Ramseier C">CA 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Int J Nanomedicine</journal-id><journal-id journal-id-type="iso-abbrev">Int J Nanomedicine</journal-id><journal-id journal-id-type="publisher-id">International Journal of Nanomedicine</journal-id><journal-title-group><journal-title>International Journal of Nanomedicine</journal-title></journal-title-group><issn pub-type="ppub">1176-9114</issn><issn pub-type="epub">1178-2013</issn><publisher><publisher-name>Dove Medical Press</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">25336945</article-id><article-id pub-id-type="pmc">4200022</article-id><article-id pub-id-type="doi">10.2147/IJN.S68895</article-id><article-id pub-id-type="publisher-id">ijn-9-4649</article-id><article-categories><subj-group subj-group-type="heading"><subject>Original Research</subject></subj-group></article-categories><title-group><article-title>Immobilization of chitosan film containing semaphorin 3A onto a microarc oxidized titanium implant surface via silane reaction to improve MG63 osteogenic differentiation</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Fang</surname><given-names>Kaixiu</given-names></name><xref ref-type="aff" rid="af1-ijn-9-4649">1</xref><xref ref-type="author-notes" rid="fn1-ijn-9-4649">*</xref></contrib><contrib contrib-type="author"><name><surname>Song</surname><given-names>Wen</given-names></name><xref ref-type="aff" rid="af2-ijn-9-4649">2</xref><xref ref-type="author-notes" rid="fn1-ijn-9-4649">*</xref></contrib><contrib contrib-type="author"><name><surname>Wang</surname><given-names>Lifeng</given-names></name><xref ref-type="aff" rid="af1-ijn-9-4649">1</xref></contrib><contrib contrib-type="author"><name><surname>Jia</surname><given-names>Sen</given-names></name><xref ref-type="aff" rid="af3-ijn-9-4649">3</xref></contrib><contrib contrib-type="author"><name><surname>Wei</surname><given-names>Hongbo</given-names></name><xref ref-type="aff" rid="af1-ijn-9-4649">1</xref></contrib><contrib contrib-type="author"><name><surname>Ren</surname><given-names>Shuai</given-names></name><xref ref-type="aff" rid="af1-ijn-9-4649">1</xref></contrib><contrib contrib-type="author"><name><surname>Xu</surname><given-names>Xiaoru</given-names></name><xref ref-type="aff" rid="af1-ijn-9-4649">1</xref></contrib><contrib contrib-type="author"><name><surname>Song</surname><given-names>Yingliang</given-names></name><xref ref-type="aff" rid="af1-ijn-9-4649">1</xref><xref ref-type="corresp" rid="c1-ijn-9-4649"></xref></contrib></contrib-group><aff id="af1-ijn-9-4649"><label>1</label>State Key Laboratory of Military Stomatology, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</aff><aff id="af2-ijn-9-4649"><label>2</label>State Key Laboratory of Military Stomatology, Department of Prosthetic Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</aff><aff id="af3-ijn-9-4649"><label>3</label>State Key Laboratory of Military Stomatology, Department of Oral and Maxillofacial Surgery, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</aff><author-notes><corresp id="c1-ijn-9-4649">Correspondence: Yingliang Song, Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, 145 West Changle Road, Xi’an 710032, People’s Republic of China, Email <email>songyingliang@yahoo.com</email></corresp><fn id="fn1-ijn-9-4649"><p>*These authors contributed equally to this work</p></fn></author-notes><pub-date pub-type="collection"><year>2014</year></pub-date><pub-date pub-type="epub"><day>03</day><month>10</month><year>2014</year></pub-date><volume>9</volume><fpage>4649</fpage><lpage>4657</lpage><permissions><copyright-statement>© 2014 Fang et al. This work is published by Dove Medical Press Limited, and licensed under Creative Commons Attribution – Non Commercial (unported, v3.0) License</copyright-statement><copyright-year>2014</copyright-year><license><license-p>The full terms of the License are available at <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by-nc/3.0/">http://creativecommons.org/licenses/by-nc/3.0/</ext-link>. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed.</license-p></license></permissions><abstract><p>Improving osseointegration of extensively used titanium (Ti) implants still remains a main theme in implantology. Recently, grafting biomolecules onto a Ti surface has attracted more attention due to their direct participation in the osseointegration process around the implant. Semaphorin 3A (Sema3A) is a new proven osteoprotection molecule and is considered to be a promising therapeutic agent in bone diseases, but how to immobilize the protein onto a Ti surface to acquire a long-term effect is poorly defined. In our study, we tried to use chitosan to wrap Sema3A (CS/Sema) and connect to the microarc oxidized Ti surface via silane glutaraldehyde coupling. The microarc oxidization could formulate porous topography on a Ti surface, and the covalently bonded coating was homogeneously covered on the ridges between the pores without significant influence on the original topography. A burst release of Sema3A was observed in the first few days in phosphate-buffered saline and could be maintained for >2 weeks. Coating in phosphate-buffered saline containing lysozyme was similar, but the release rate was much more rapid. The coating did not significantly affect cellular adhesion, viability, or cytoskeleton arrangement, but the osteogenic-related gene expression was dramatically increased and calcium deposition was also abundantly detected. In conclusion, covalent bonding of CS/Sema could strongly improve osteogenic differentiation of osteoblasts and might be applied for Ti implant surface biofunctionalization.</p></abstract><kwd-group><title>Keywords</title><kwd>titanium</kwd><kwd>semaphorin 3A</kwd><kwd>silane reaction</kwd><kwd>microarc oxidation</kwd><kwd>osteogenic differentiation</kwd></kwd-group></article-meta></front><floats-group><fig id="f1-ijn-9-4649" position="float"><label>Figure 1</label><caption><p>The coating morphology observation by scanning electron microscopy (<bold>A</bold>) and water contact angle measurement (<bold>B</bold>).</p><p><bold>Note:</bold> *<italic>P</italic><0.05 vs MAO.</p><p><bold>Abbreviations:</bold> CS/Sema, chitosan–semaphorin 3A; MAO, microarc oxidation.</p></caption><graphic xlink:href="ijn-9-4649Fig1"></graphic></fig><fig id="f2-ijn-9-4649" position="float"><label>Figure 2</label><caption><p>Accumulated release profile of CS/Sema–MAO in PBS and PBS + lysozyme (<bold>A</bold>) and the morphology observation of the substrate after 1 day (<bold>B1</bold>), 3 days (<bold>B2</bold>), 7 days (<bold>B3</bold>), and 14 days (<bold>B4</bold>) of dissolution in PBS.</p><p><bold>Abbreviations:</bold> CS/Sema, chitosan–semaphorin 3A; MAO, microarc oxidation; PBS, phosphate-buffered saline.</p></caption><graphic xlink:href="ijn-9-4649Fig2"></graphic></fig><fig id="f3-ijn-9-4649" position="float"><label>Figure 3</label><caption><p>Cell adhesion visualized by 4′,6-diamidino-2-phenylindole staining (<bold>A</bold>), scanning electron microscope (<bold>B</bold>) and number counts (<bold>C</bold>).</p><p><bold>Notes:</bold> (<bold>A</bold>)(<bold>a</bold>) and (<bold>B</bold>)(<bold>a</bold>): CS/Sema-MAO; (<bold>A</bold>)(<bold>b</bold>) and (<bold>B</bold>)(<bold>b</bold>): CS/BSA-MAO; (<bold>A</bold>)(<bold>c</bold>) and (<bold>B</bold>)(<bold>c</bold>): CS-MAO; (<bold>A</bold>)(<bold>d</bold>) and (<bold>B</bold>)(<bold>d</bold>): MAO.</p><p><bold>Abbreviations:</bold> CS/Sema, chitosan–semaphorin 3A; MAO, microarc oxidation; CS/BSA, chitosan-bovine serum albumin.</p></caption><graphic xlink:href="ijn-9-4649Fig3"></graphic></fig><fig id="f4-ijn-9-4649" position="float"><label>Figure 4</label><caption><p>Cellular viability carried out by 3-(4,5-dimethylthiazol-2yl)-2, 5-diphenyltetrazolium bromide assay.</p><p><bold>Abbreviations:</bold> CS/Sema, chitosan–semaphorin 3A; MAO, microarc oxidation; CS/BSA, chitosan-bovine serum albumin.</p></caption><graphic xlink:href="ijn-9-4649Fig4"></graphic></fig><fig id="f5-ijn-9-4649" position="float"><label>Figure 5</label><caption><p>Cytoskeleton staining by rhodamine phalloidin under confocal laser scanning microscopy observation.</p><p><bold>Notes:</bold> A: CS/Sema–MAO; B: CS/BSA–MAO; C: CS–MAO; D: MAO; scale bar =100 μm.</p><p><bold>Abbreviations:</bold> CS/Sema, chitosan–semaphorin 3A; MAO, microarc oxidation.</p></caption><graphic xlink:href="ijn-9-4649Fig5"></graphic></fig><fig id="f6-ijn-9-4649" position="float"><label>Figure 6</label><caption><p>Osteogenic-related gene expression quantified by real-time quantitative polymerase chain reaction after osteogenic induction of 3 days (<bold>A</bold>) and 7 days (<bold>B</bold>). The relative protein level after 3 days of culture was analyzed with Western blot (<bold>C</bold>).</p><p><bold>Notes:</bold> *<italic>P</italic><0.05 vs CS/BSA–MAO, CS–MAO, and MAO.</p><p><bold>Abbreviations:</bold> CS/Sema, chitosan–semaphorin 3A; MAO, microarc oxidation; RUNX2, runt-related transcription factor 2; ALP, alkaline phosphatase; OCN, osteocalcin; BMP, bone morphogenetic protein; CS/BSA, chitosan-bovine serum albumin; mRNA, messenger RNA.</p></caption><graphic xlink:href="ijn-9-4649Fig6"></graphic></fig><fig id="f7-ijn-9-4649" position="float"><label>Figure 7</label><caption><p>Extracellular matrix mineralization stained by Alizarin Red Solution (<bold>A</bold>) and optical density measurement (<bold>B</bold>).</p><p><bold>Notes:</bold> *<italic>P</italic><0.05 vs CS/BSA–MAO, CS–MAO, and MAO.</p><p><bold>Abbreviations:</bold> CS/Sema, chitosan–semaphorin 3A; MAO, microarc oxidation; CS/BSA, chitosan-bovine serum albumin.</p></caption><graphic xlink:href="ijn-9-4649Fig7"></graphic></fig><table-wrap id="t1-ijn-9-4649" position="float"><label>Table 1</label><caption><p>Primers used for real-time quantitative polymerase chain reaction</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" valign="top" rowspan="1" colspan="1">Gene</th><th align="left" valign="top" rowspan="1" colspan="1">Forward primer sequence (5′-3′)</th><th align="left" valign="top" rowspan="1" colspan="1">Reverse primer sequence (5′-3′)</th></tr></thead><tbody><tr><td align="left" valign="top" rowspan="1" colspan="1"><italic>RUNX2</italic></td><td align="left" valign="top" rowspan="1" colspan="1">CACTGGCGCTGCAACAAGA</td><td align="left" valign="top" rowspan="1" colspan="1">CATTCCGGAGCTCAGCAGAATAA</td></tr><tr><td align="left" valign="top" rowspan="1" colspan="1"><italic>ALP</italic></td><td align="left" valign="top" rowspan="1" colspan="1">CCTTGTAGCCAGGCCCATTG</td><td align="left" valign="top" rowspan="1" colspan="1">GGACCATTCCCACGTCTTCAC</td></tr><tr><td align="left" valign="top" rowspan="1" colspan="1"><italic>OCN</italic></td><td align="left" valign="top" rowspan="1" colspan="1">CCCAGGCGCTACCTGTATCAA</td><td align="left" valign="top" rowspan="1" colspan="1">GGTCAGCCAACTCGTCACAGTC</td></tr><tr><td align="left" valign="top" rowspan="1" colspan="1"><italic>BMP</italic></td><td align="left" valign="top" rowspan="1" colspan="1">CAACACCGTGCTCAGCTTCC</td><td align="left" valign="top" rowspan="1" colspan="1">TTCCCACTCATTTCTGAAAGTTCC</td></tr><tr><td align="left" valign="top" rowspan="1" colspan="1"><italic>β-actin</italic></td><td align="left" valign="top" rowspan="1" colspan="1">TGGCACCCAGCACAATGAA</td><td align="left" valign="top" rowspan="1" colspan="1">CTAAGTCATAGTCCGCCTAGAAGCA</td></tr></tbody></table><table-wrap-foot><fn id="tfn1-ijn-9-4649"><p><bold>Abbreviations:</bold> RUNX2, runt-related transcription factor 2; ALP, alkaline phosphatase; OCN, osteocalcin; BMP, bone morphogenetic protein.</p></fn></table-wrap-foot></table-wrap></floats-group></pmc><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Fang, Kaixiu" sort="Fang, Kaixiu" uniqKey="Fang K" first="Kaixiu" last="Fang">Kaixiu Fang</name></noRegion><name sortKey="Jia, Sen" sort="Jia, Sen" uniqKey="Jia S" first="Sen" last="Jia">Sen Jia</name><name sortKey="Ren, Shuai" sort="Ren, Shuai" uniqKey="Ren S" first="Shuai" last="Ren">Shuai Ren</name><name sortKey="Song, Wen" sort="Song, Wen" uniqKey="Song W" first="Wen" last="Song">Wen Song</name><name sortKey="Song, Yingliang" sort="Song, Yingliang" uniqKey="Song Y" first="Yingliang" last="Song">Yingliang Song</name><name sortKey="Wang, Lifeng" sort="Wang, Lifeng" uniqKey="Wang L" first="Lifeng" last="Wang">Lifeng Wang</name><name sortKey="Wei, Hongbo" sort="Wei, Hongbo" uniqKey="Wei H" first="Hongbo" last="Wei">Hongbo Wei</name><name sortKey="Xu, Xiaoru" sort="Xu, Xiaoru" uniqKey="Xu X" first="Xiaoru" last="Xu">Xiaoru Xu</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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