Concentration- and time-dependent response of human gingival fibroblasts to fibroblast growth factor 2 immobilized on titanium dental implants
Identifieur interne : 001F29 ( Pmc/Checkpoint ); précédent : 001F28; suivant : 001F30Concentration- and time-dependent response of human gingival fibroblasts to fibroblast growth factor 2 immobilized on titanium dental implants
Auteurs : Qianli Ma [République populaire de Chine] ; Wei Wang [République populaire de Chine] ; Paul K. Chu [République populaire de Chine] ; Shenglin Mei [République populaire de Chine] ; Kun Ji [République populaire de Chine] ; Lei Jin [République populaire de Chine] ; Yumei Zhang [République populaire de Chine]Source :
- International Journal of Nanomedicine [ 1176-9114 ] ; 2012.
Abstract
Titanium (Ti) implants are widely used clinically, but peri-implantitis remains one of the most common and serious complications. Healthy integration between gingival tissue and the implant surface is critical to long-term success in dental implant therapy. The objective of this study was to investigate how different concentrations of immobilized fibroblast growth factor 2 (FGF2) on the titania nanotubular surface influence the response of human gingival fibroblasts (HGFs).
Pure Ti metal was anodized at 20 V to form a vertically organized titanium dioxide nanotube array on which three concentrations of FGF2 (250 ng/mL, 500 ng/mL, or 1000 ng/mL) were immobilized by repeated lyophilization. Surface topography was observed and FGF2 elution was detected using enzyme-linked immunosorbent assay. The bioactivity changes of dissolvable immobilized FGF2 were measured by methyl-thiazolyl-tetrazolium assay. Behavior of HGFs was evaluated using adhesion and methyl-thiazolyl-tetrazolium bromide assays.
The FGF2 remained for several days on the modified surface on which HGFs were cultured. Over 90% of the dissolvable immobilized FGF2 had been eluted by Day 9, whereas the FGF2 activity was found to diminish gradually from Day 1 to Day 9. The titania nanotubular surface with an optimal preparing concentration (500 ng/mL) of FGF2 immobilization exhibited improved HGF functions such as cellular attachment, proliferation, and extracellular matrix-related gene expression. Moreover, significant bidirectional as well as concentration- and time-dependent bioactivity was observed.
Synergism of the FGF2-impregnated titanium dioxide nanotubular surface revealed good gingival-implant integration, indicating that these materials might have promising applications in dentistry and other biomedical devices.
Url:
DOI: 10.2147/IJN.S29538
PubMed: 22619534
PubMed Central: 3356224
Affiliations:
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<front><div type="abstract" xml:lang="en"><sec><title>Background</title>
<p>Titanium (Ti) implants are widely used clinically, but peri-implantitis remains one of the most common and serious complications. Healthy integration between gingival tissue and the implant surface is critical to long-term success in dental implant therapy. The objective of this study was to investigate how different concentrations of immobilized fibroblast growth factor 2 (FGF2) on the titania nanotubular surface influence the response of human gingival fibroblasts (HGFs).</p>
</sec>
<sec><title>Methods</title>
<p>Pure Ti metal was anodized at 20 V to form a vertically organized titanium dioxide nanotube array on which three concentrations of FGF2 (250 ng/mL, 500 ng/mL, or 1000 ng/mL) were immobilized by repeated lyophilization. Surface topography was observed and FGF2 elution was detected using enzyme-linked immunosorbent assay. The bioactivity changes of dissolvable immobilized FGF2 were measured by methyl-thiazolyl-tetrazolium assay. Behavior of HGFs was evaluated using adhesion and methyl-thiazolyl-tetrazolium bromide assays.</p>
</sec>
<sec><title>Results</title>
<p>The FGF2 remained for several days on the modified surface on which HGFs were cultured. Over 90% of the dissolvable immobilized FGF2 had been eluted by Day 9, whereas the FGF2 activity was found to diminish gradually from Day 1 to Day 9. The titania nanotubular surface with an optimal preparing concentration (500 ng/mL) of FGF2 immobilization exhibited improved HGF functions such as cellular attachment, proliferation, and extracellular matrix-related gene expression. Moreover, significant bidirectional as well as concentration- and time-dependent bioactivity was observed.</p>
</sec>
<sec><title>Conclusion</title>
<p>Synergism of the FGF2-impregnated titanium dioxide nanotubular surface revealed good gingival-implant integration, indicating that these materials might have promising applications in dentistry and other biomedical devices.</p>
</sec>
</div>
</front>
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</TEI>
<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<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-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">22619534</article-id>
<article-id pub-id-type="pmc">3356224</article-id>
<article-id pub-id-type="doi">10.2147/IJN.S29538</article-id>
<article-id pub-id-type="publisher-id">ijn-7-1965</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Original Research</subject>
</subj-group>
</article-categories>
<title-group><article-title>Concentration- and time-dependent response of human gingival fibroblasts to fibroblast growth factor 2 immobilized on titanium dental implants</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Ma</surname>
<given-names>Qianli</given-names>
</name>
<xref ref-type="aff" rid="af1-ijn-7-1965">1</xref>
<xref ref-type="author-notes" rid="fn1-ijn-7-1965">*</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Wang</surname>
<given-names>Wei</given-names>
</name>
<xref ref-type="aff" rid="af1-ijn-7-1965">1</xref>
<xref ref-type="author-notes" rid="fn1-ijn-7-1965">*</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Chu</surname>
<given-names>Paul K</given-names>
</name>
<xref ref-type="aff" rid="af2-ijn-7-1965">2</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Mei</surname>
<given-names>Shenglin</given-names>
</name>
<xref ref-type="aff" rid="af1-ijn-7-1965">1</xref>
<xref ref-type="aff" rid="af2-ijn-7-1965">2</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Ji</surname>
<given-names>Kun</given-names>
</name>
<xref ref-type="aff" rid="af3-ijn-7-1965">3</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Jin</surname>
<given-names>Lei</given-names>
</name>
<xref ref-type="aff" rid="af4-ijn-7-1965">4</xref>
<xref ref-type="corresp" rid="c1-ijn-7-1965"></xref>
</contrib>
<contrib contrib-type="author"><name><surname>Zhang</surname>
<given-names>Yumei</given-names>
</name>
<xref ref-type="aff" rid="af1-ijn-7-1965">1</xref>
<xref ref-type="corresp" rid="c1-ijn-7-1965"></xref>
</contrib>
</contrib-group>
<aff id="af1-ijn-7-1965"><label>1</label>
Department of Prosthetic Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</aff>
<aff id="af2-ijn-7-1965"><label>2</label>
Department of Physics and Materials Science, City University of Hong Kong, Kowloon, Hong Kong, People’s Republic of China</aff>
<aff id="af3-ijn-7-1965"><label>3</label>
Department of Pediatric Dentistry, School of Stomatology, Fourth Military Medical University, Xi’an, People’s Republic of China</aff>
<aff id="af4-ijn-7-1965"><label>4</label>
Stomatology Department, Jinling Hospital, School of Medicine, Southern Medical University, Nanjing, People’s Republic of China</aff>
<author-notes><corresp id="c1-ijn-7-1965">Correspondence: Yumei Zhang, Department of Prosthetic Dentistry, School of Stomatology, Fourth Military Medical University, 17 West Chang Le Road, Xi’an 710032, People’s Republic of China, Tel +86 29 84776090, Fax +86 29 84776096, Email <email>wqtzym@fmmu.edu.cn</email>
. Lei Jin, Stomatology Department, Jinling Hospital, School of Medicine, Southern Medical University, 305 East Zhongshan Road, Nanjing, 210002, People’s Republic of China, Tel +86 25 80861166, Fax +86 25 80861166, Email <email>jindentist@gmail.com</email>
</corresp>
<fn id="fn1-ijn-7-1965"><label>*</label>
<p>These authors contributed equally to this work</p>
</fn>
</author-notes>
<pub-date pub-type="collection"><year>2012</year>
</pub-date>
<pmc-comment>Dove Press titles changed from ppub to collections in 2009.
Fake ppub written to satisfy Coll Date Type=ppub</pmc-comment>
<pub-date pub-type="ppub"><year>2012</year>
</pub-date>
<pub-date pub-type="epub"><day>17</day>
<month>4</month>
<year>2012</year>
</pub-date>
<volume>7</volume>
<fpage>1965</fpage>
<lpage>1976</lpage>
<permissions><copyright-statement>© 2012 Ma et al, publisher and licensee Dove Medical Press Ltd.</copyright-statement>
<copyright-year>2012</copyright-year>
<license><license-p>This is an Open Access article which permits unrestricted noncommercial use, provided the original work is properly cited.</license-p>
</license>
</permissions>
<abstract><sec><title>Background</title>
<p>Titanium (Ti) implants are widely used clinically, but peri-implantitis remains one of the most common and serious complications. Healthy integration between gingival tissue and the implant surface is critical to long-term success in dental implant therapy. The objective of this study was to investigate how different concentrations of immobilized fibroblast growth factor 2 (FGF2) on the titania nanotubular surface influence the response of human gingival fibroblasts (HGFs).</p>
</sec>
<sec><title>Methods</title>
<p>Pure Ti metal was anodized at 20 V to form a vertically organized titanium dioxide nanotube array on which three concentrations of FGF2 (250 ng/mL, 500 ng/mL, or 1000 ng/mL) were immobilized by repeated lyophilization. Surface topography was observed and FGF2 elution was detected using enzyme-linked immunosorbent assay. The bioactivity changes of dissolvable immobilized FGF2 were measured by methyl-thiazolyl-tetrazolium assay. Behavior of HGFs was evaluated using adhesion and methyl-thiazolyl-tetrazolium bromide assays.</p>
</sec>
<sec><title>Results</title>
<p>The FGF2 remained for several days on the modified surface on which HGFs were cultured. Over 90% of the dissolvable immobilized FGF2 had been eluted by Day 9, whereas the FGF2 activity was found to diminish gradually from Day 1 to Day 9. The titania nanotubular surface with an optimal preparing concentration (500 ng/mL) of FGF2 immobilization exhibited improved HGF functions such as cellular attachment, proliferation, and extracellular matrix-related gene expression. Moreover, significant bidirectional as well as concentration- and time-dependent bioactivity was observed.</p>
</sec>
<sec><title>Conclusion</title>
<p>Synergism of the FGF2-impregnated titanium dioxide nanotubular surface revealed good gingival-implant integration, indicating that these materials might have promising applications in dentistry and other biomedical devices.</p>
</sec>
</abstract>
<kwd-group><kwd>dental implants</kwd>
<kwd>titanium dioxide nanotube</kwd>
<kwd>fibroblast growth factor 2</kwd>
<kwd>extracellular matrix</kwd>
<kwd>real-time polymerase chain reaction</kwd>
</kwd-group>
</article-meta>
</front>
<floats-group><fig id="f1-ijn-7-1965" position="float"><label>Figure 1</label>
<caption><p>Immobilized FGF2 compared with FGF2-free samples initially and after 9 days.</p>
<p><bold>Abbreviations:</bold>
FGF2, fibroblast growth factor 2; NT, nanotube; PT, polished titanium.</p>
</caption>
<graphic xlink:href="ijn-7-1965f1"></graphic>
</fig>
<fig id="f2-ijn-7-1965" position="float"><label>Figure 2</label>
<caption><p>Field emission scanning electron microscopy results. (<bold>A</bold>
) NT surface showed an NT array structure; NT-F-H surface showed clumps of fibroblast growth factor 2 and that the typical NT surface was changed. (<bold>B</bold>
) Profile of titania NT.</p>
<p><bold>Note:</bold>
The length of NT was 588.85 ± 31.92 nm (mean value ± standard deviation).</p>
<p><bold>Abbreviations:</bold>
NT, nanotube; PT, polished titanium.</p>
</caption>
<graphic xlink:href="ijn-7-1965f2"></graphic>
</fig>
<fig id="f3-ijn-7-1965" position="float"><label>Figure 3</label>
<caption><p>Atomic force microscopy images of different surfaces: (<bold>A</bold>
) PT, (<bold>B</bold>
) NT, (<bold>C</bold>
) NT-F-L, (<bold>D</bold>
) NT-F-M, and (<bold>E</bold>
) NT-F-H. PT showed a microgroove structure, and porous-like structures were observed in other groups.</p>
<p><bold>Abbreviations:</bold>
NT, nanotube; PT, polished titanium.</p>
</caption>
<graphic xlink:href="ijn-7-1965f3"></graphic>
</fig>
<fig id="f4-ijn-7-1965" position="float"><label>Figure 4</label>
<caption><p>Elution kinetics of various FGF2 immobilizations. FGF2 eluted rapidly within first 3 days and the elution velocity reduced with time. Approximately all dissolvable FGF2 eluted within 30 days.</p>
<p><bold>Abbreviations:</bold>
FGF2, fibroblast growth factor 2; NT, nanotube.</p>
</caption>
<graphic xlink:href="ijn-7-1965f4"></graphic>
</fig>
<fig id="f5-ijn-7-1965" position="float"><label>Figure 5</label>
<caption><p>HGF adhesion. (<bold>A</bold>
) HGFs attached to the substrates with 4′,6′-diamidino-2-phenylindole stain and (<bold>B</bold>
) relative cell adhesion rate of attached HGFs. The NT and NT-F-H surfaces inhibited cell adhesion, whereas the NT-F-L and NT-F-M improved cell attachment.</p>
<p><bold>Notes:</bold>
<sup>#</sup>
Higher than PT (<italic>P</italic>
< 0.01), *lower than PT (<italic>P</italic>
< 0.01), difference also exists between 1*, 2*, 1<sup>#</sup>
, and 2<sup>#</sup>
(<italic>P</italic>
< 0.01).</p>
<p><bold>Abbreviations:</bold>
HGF, human gingival fibroblast; NT, nanotube; PT, polished titanium.</p>
</caption>
<graphic xlink:href="ijn-7-1965f5"></graphic>
</fig>
<fig id="f6-ijn-7-1965" position="float"><label>Figure 6</label>
<caption><p>Cell proliferation measured by the methyl-thiazolyl-tetrazolium assay. NT-F-L and NT-F-M showed the higher cell proliferation at each interval, whereas NT-F-H with overimmobilized FGF2 showed the lowest at the early stage (Days 1 and 3).</p>
<p><bold>Notes:</bold>
<sup>#</sup>
Higher than PT (<italic>P</italic>
< 0.01), *lower than PT (<italic>P</italic>
< 0.01), significant difference exists between 1<sup>#</sup>
and 2<sup>#</sup>
(<italic>P</italic>
< 0.01).</p>
<p><bold>Abbreviations:</bold>
FGF2, fibroblast growth factor 2; NT, nanotube; OD, optical density; PT, polished titanium.</p>
</caption>
<graphic xlink:href="ijn-7-1965f6"></graphic>
</fig>
<fig id="f7-ijn-7-1965" position="float"><label>Figure 7</label>
<caption><p>Morphology of HGFs: (<bold>A</bold>
) PT, (<bold>B</bold>
) NT, (<bold>C</bold>
) NT-F-L, (<bold>D</bold>
) NT-F-M, and (<bold>E</bold>
) NT-F-H lacking sufficient prominences/pseudopods and not fusing together.</p>
<p><bold>Abbreviations:</bold>
HGF, human gingival fibroblast; NT, nanotube; PT, polished titanium.</p>
</caption>
<graphic xlink:href="ijn-7-1965f7"></graphic>
</fig>
<fig id="f8-ijn-7-1965" position="float"><label>Figure 8</label>
<caption><p>Extracellular matrix-related gene expressions by HGFs cultured on four different titanium surfaces at Days 3, 6, and 9: (<bold>A</bold>
) <italic>VEGFA</italic>
, (<bold>B</bold>
) <italic>ITGB</italic>
, (<bold>C</bold>
) <italic>ICAM1</italic>
, and (<bold>D</bold>
) <italic>LAMA1</italic>
. Inverse concentration-dependent effects of NT-F-H were observed at Day 3 on (<bold>C</bold>
) and (<bold>D</bold>
).</p>
<p><bold>Notes:</bold>
<sup>#</sup>
Upregulated compared with PT (<italic>P</italic>
< 0.01), *downregulated (<italic>P</italic>
< 0.01), significant difference exists between 1*, 2*, 1<sup>#</sup>
, and 2<sup>#</sup>
(<italic>P</italic>
< 0.01).</p>
<p><bold>Abbreviations:</bold>
HGF, human gingival fibroblast; <italic>ICAM1</italic>
, intercellular adhesion molecule 1; <italic>ITGB</italic>
, β-integrin; <italic>LAMA1</italic>
, laminin-1; NT, nanotube; PT, polished titanium; <italic>VEGFA,</italic>
vascular endothelial growth factor A.</p>
</caption>
<graphic xlink:href="ijn-7-1965f8"></graphic>
</fig>
<fig id="f9-ijn-7-1965" position="float"><label>Figure 9</label>
<caption><p>Activity changes of dissolvable FGF2 (10 ng/mL): control (FGF2-free); newly prepared Dulbecco’s modified Eagle’s medium; and S1, S3, S6, and S9 (FGF2 in Dulbecco’s modified Eagle’s medium stored for 1 day, 3 days, 6 days, or 9 days, respectively). The activity of FGF2 reduced gradually in culture medium with time and dropped to less than 50% after 9 days (S9) compared with NP.</p>
<p><bold>Note:</bold>
<sup>#</sup>
Higher than PT (<italic>P</italic>
< 0.01), significant difference exists between 1<sup>#</sup>
, 2<sup>#</sup>
, and 3<sup>#</sup>
(<italic>P</italic>
< 0.01).</p>
<p><bold>Abbreviations:</bold>
FGF2, fibroblast growth factor 2; NT, nanotube; OD, optical density; PT, polished titanium.</p>
</caption>
<graphic xlink:href="ijn-7-1965f9"></graphic>
</fig>
<fig id="f10-ijn-7-1965" position="float"><label>Figure 10</label>
<caption><p>Pattern of nanotube-based FGF2 control release system: (<bold>A</bold>
) initial stage, (<bold>B</bold>
) in culture medium, FGF2 is gradually released from the nanotubes.</p>
<p><bold>Abbreviation:</bold>
FGF2, fibroblast growth factor 2.</p>
</caption>
<graphic xlink:href="ijn-7-1965f10"></graphic>
</fig>
<table-wrap id="t1-ijn-7-1965" position="float"><label>Table 1</label>
<caption><p>Target cDNA primer sequences used in quantitative polymerase chain reaction</p>
</caption>
<table frame="hsides" rules="groups"><tbody><tr><td align="left" valign="top" rowspan="1" colspan="1"><italic>VEGFA</italic>
</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> Forward: 5′-GAGCCTTGCCTTGCTGCTCTAC-3′</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> Reverse: 5′-CACCAGGTCTCGATTGGATG-3′</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> PCR product size: 148 bp</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"><italic>ITGB</italic>
</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> Forward: 5′-TGTGTCAGACCTGCCTTGGTG-3′</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> Reverse: 5′-AGGAACATTCCTGTGTGCATGTG-3′</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> PCR product size: 105 bp</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"><italic>ICAM1</italic>
</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> Forward: TGAGCAATGTGCAAGAAGATAGC</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> Reverse: CCCGTTCTGGAGTCCAGTACA</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> PCR product size: 105 bp</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"><italic>LAMA1</italic>
</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> Forward: TGGTAACATTTGCCACCACGA</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> Reverse: TTGCCTCCGATCAGCATGAC</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> PCR product size: 127 bp</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"><italic>GAPDH</italic>
(housekeeping gene)</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> Forward: 5′-GCACCGTCAAGGCTGAGAAC-3′</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> Reverse: 5′-TGGTGAAGACGCCAGTGGA-3′</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1"> PCR product size: 138 bp</td>
</tr>
</tbody>
</table>
<table-wrap-foot><fn id="tfn1-ijn-7-1965"><p><bold>Abbreviations:</bold>
<italic>GAPDH</italic>
, glyceraldehyde 3-phosphate dehydrogenase; <italic>ICAM1</italic>
, intercellular adhesion molecule 1; <italic>ITGB</italic>
, β-integrin; <italic>LAMA1</italic>
, laminin-1; <italic>VEGFA,</italic>
vascular endothelial growth factor A.</p>
</fn>
</table-wrap-foot>
</table-wrap>
<table-wrap id="t2-ijn-7-1965" position="float"><label>Table 2</label>
<caption><p>Measurement of surface roughness by atomic force microscopy</p>
</caption>
<table frame="hsides" rules="groups"><thead><tr><th align="left" valign="top" rowspan="1" colspan="1">Group</th>
<th align="left" valign="top" rowspan="1" colspan="1">R<sub>a</sub>
</th>
<th align="left" valign="top" rowspan="1" colspan="1">RMS</th>
</tr>
</thead>
<tbody><tr><td align="left" valign="top" rowspan="1" colspan="1">PT</td>
<td align="center" valign="top" rowspan="1" colspan="1">32.60 ± 3.45 nm</td>
<td align="center" valign="top" rowspan="1" colspan="1">41.70 ± 3.96 nm</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">NT</td>
<td align="center" valign="top" rowspan="1" colspan="1">4.96 ± 0.50 nm<xref ref-type="table-fn" rid="tfn3-ijn-7-1965">*</xref>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">6.23 ± 0.80 nm<xref ref-type="table-fn" rid="tfn3-ijn-7-1965">*</xref>
</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">NT-F-L</td>
<td align="center" valign="top" rowspan="1" colspan="1">7.25 ± 0.97 nm<xref ref-type="table-fn" rid="tfn3-ijn-7-1965">*</xref>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">9.14 ± 1.25 nm<xref ref-type="table-fn" rid="tfn3-ijn-7-1965">*</xref>
</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">NT-F-M</td>
<td align="center" valign="top" rowspan="1" colspan="1">9.70 ± 1.85 nm<xref ref-type="table-fn" rid="tfn3-ijn-7-1965">*</xref>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">11.80 ± 2.21 nm<xref ref-type="table-fn" rid="tfn3-ijn-7-1965">*</xref>
</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">NT-F-H</td>
<td align="center" valign="top" rowspan="1" colspan="1">9.42 ± 1.99 nm<xref ref-type="table-fn" rid="tfn3-ijn-7-1965">*</xref>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">11.79 ± 2.35 nm<xref ref-type="table-fn" rid="tfn3-ijn-7-1965">*</xref>
</td>
</tr>
</tbody>
</table>
<table-wrap-foot><fn id="tfn2-ijn-7-1965"><p><bold>Notes:</bold>
</p>
</fn>
<fn id="tfn3-ijn-7-1965"><label>*</label>
<p>Lower R<sub>a</sub>
and RMS compared with PT (<italic>P</italic>
< 0.01). Anodization significantly smoothed titanium surface, whereas surface roughness was barely affected by FGF2 immobilization.</p>
</fn>
<fn id="tfn4-ijn-7-1965"><p><bold>Abbreviations:</bold>
FGF2, fibroblast growth factor 2; NT, nanotube; PT, polished titanium; R<sub>a</sub>
, average roughness, RMS, root mean square roughness.</p>
</fn>
</table-wrap-foot>
</table-wrap>
<table-wrap id="t3-ijn-7-1965" position="float"><label>Table 3</label>
<caption><p>Elution of dissolvable FGF2 at different time intervals<xref ref-type="table-fn" rid="tfn6-ijn-7-1965">*</xref>
</p>
</caption>
<table frame="hsides" rules="groups"><thead><tr><th align="left" valign="top" rowspan="1" colspan="1">Time interval</th>
<th colspan="2" align="left" valign="top" rowspan="1">NT-F-L</th>
<th colspan="2" align="left" valign="top" rowspan="1">NT-F-M</th>
<th colspan="2" align="left" valign="top" rowspan="1">NT-F-H</th>
</tr>
<tr><th align="left" valign="top" rowspan="1" colspan="1"></th>
<th colspan="2" align="left" valign="top" rowspan="1"><hr></hr>
</th>
<th colspan="2" align="left" valign="top" rowspan="1"><hr></hr>
</th>
<th colspan="2" align="left" valign="top" rowspan="1"><hr></hr>
</th>
</tr>
<tr><th align="left" valign="top" rowspan="1" colspan="1"></th>
<th align="left" valign="top" rowspan="1" colspan="1">Mass (ng)</th>
<th align="left" valign="top" rowspan="1" colspan="1">Percent (accumulative)</th>
<th align="left" valign="top" rowspan="1" colspan="1">Mass (ng)</th>
<th align="left" valign="top" rowspan="1" colspan="1">Percent (accumulative)</th>
<th align="left" valign="top" rowspan="1" colspan="1">Mass (ng)</th>
<th align="left" valign="top" rowspan="1" colspan="1">Percent (accumulative)</th>
</tr>
</thead>
<tbody><tr><td align="left" valign="top" rowspan="1" colspan="1">D0~D1</td>
<td align="left" valign="top" rowspan="1" colspan="1">10</td>
<td align="left" valign="top" rowspan="1" colspan="1">36.76</td>
<td align="left" valign="top" rowspan="1" colspan="1">24</td>
<td align="left" valign="top" rowspan="1" colspan="1">36.70</td>
<td align="left" valign="top" rowspan="1" colspan="1">35.6</td>
<td align="left" valign="top" rowspan="1" colspan="1">41.59</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">D1~D2</td>
<td align="left" valign="top" rowspan="1" colspan="1">6</td>
<td align="left" valign="top" rowspan="1" colspan="1">58.82</td>
<td align="left" valign="top" rowspan="1" colspan="1">14.1</td>
<td align="left" valign="top" rowspan="1" colspan="1">58.26</td>
<td align="left" valign="top" rowspan="1" colspan="1">16.5</td>
<td align="left" valign="top" rowspan="1" colspan="1">60.86</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">D2~D3</td>
<td align="left" valign="top" rowspan="1" colspan="1">3</td>
<td align="left" valign="top" rowspan="1" colspan="1">69.85</td>
<td align="left" valign="top" rowspan="1" colspan="1">7.9</td>
<td align="left" valign="top" rowspan="1" colspan="1">70.34</td>
<td align="left" valign="top" rowspan="1" colspan="1">8.9</td>
<td align="left" valign="top" rowspan="1" colspan="1">71.26</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">D3~D6</td>
<td align="left" valign="top" rowspan="1" colspan="1">4.5</td>
<td align="left" valign="top" rowspan="1" colspan="1">86.40</td>
<td align="left" valign="top" rowspan="1" colspan="1">11.5</td>
<td align="left" valign="top" rowspan="1" colspan="1">87.92</td>
<td align="left" valign="top" rowspan="1" colspan="1">15.5</td>
<td align="left" valign="top" rowspan="1" colspan="1">89.37</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">D6~D9</td>
<td align="left" valign="top" rowspan="1" colspan="1">1.5</td>
<td align="left" valign="top" rowspan="1" colspan="1">91.91</td>
<td align="left" valign="top" rowspan="1" colspan="1">4.5</td>
<td align="left" valign="top" rowspan="1" colspan="1">94.80</td>
<td align="left" valign="top" rowspan="1" colspan="1">5.7</td>
<td align="left" valign="top" rowspan="1" colspan="1">96.03</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">D9~D15</td>
<td align="left" valign="top" rowspan="1" colspan="1">1.4</td>
<td align="left" valign="top" rowspan="1" colspan="1">97.06</td>
<td align="left" valign="top" rowspan="1" colspan="1">2.3</td>
<td align="left" valign="top" rowspan="1" colspan="1">98.32</td>
<td align="left" valign="top" rowspan="1" colspan="1">2.1</td>
<td align="left" valign="top" rowspan="1" colspan="1">98.48</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">D15~D21</td>
<td align="left" valign="top" rowspan="1" colspan="1">0.6</td>
<td align="left" valign="top" rowspan="1" colspan="1">99.26</td>
<td align="left" valign="top" rowspan="1" colspan="1">0.7</td>
<td align="left" valign="top" rowspan="1" colspan="1">99.39</td>
<td align="left" valign="top" rowspan="1" colspan="1">0.8</td>
<td align="left" valign="top" rowspan="1" colspan="1">99.42</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">D21~D30</td>
<td align="left" valign="top" rowspan="1" colspan="1">0.2</td>
<td align="left" valign="top" rowspan="1" colspan="1">100.00</td>
<td align="left" valign="top" rowspan="1" colspan="1">0.4</td>
<td align="left" valign="top" rowspan="1" colspan="1">100.00</td>
<td align="left" valign="top" rowspan="1" colspan="1">0.5</td>
<td align="left" valign="top" rowspan="1" colspan="1">100.00</td>
</tr>
</tbody>
</table>
<table-wrap-foot><fn id="tfn5-ijn-7-1965"><p><bold>Notes:</bold>
</p>
</fn>
<fn id="tfn6-ijn-7-1965"><label>*</label>
<p>Average amounts and accumulative percent of eluted FGF2 at each time interval. FGF2 rapidly eluted within the first 3 days, followed by a gradual reduction in elution. Over 90% of the dissolvable immobilized FGF2 eluted within 9 days. After 21 days, additional elution was negligible.</p>
</fn>
<fn id="tfn7-ijn-7-1965"><p><bold>Abbreviations:</bold>
FGF2, fibroblast growth factor 2; NT, nanotube; PT, polished titanium.</p>
</fn>
</table-wrap-foot>
</table-wrap>
<table-wrap id="t4-ijn-7-1965" position="float"><label>Table 4</label>
<caption><p>Summary of extracellular matrix-related gene expressions on Days 3, 6, and 9</p>
</caption>
<table frame="hsides" rules="groups"><thead><tr><th align="left" valign="top" rowspan="1" colspan="1"></th>
<th colspan="3" align="left" valign="top" rowspan="1"><italic>VEGFA</italic>
</th>
<th colspan="3" align="left" valign="top" rowspan="1"><italic>ITGB</italic>
</th>
<th colspan="3" align="left" valign="top" rowspan="1"><italic>ICAM1</italic>
</th>
<th colspan="3" align="left" valign="top" rowspan="1"><italic>LAMA1</italic>
</th>
</tr>
<tr><th align="left" valign="top" rowspan="1" colspan="1"></th>
<th colspan="3" align="left" valign="top" rowspan="1"><hr></hr>
</th>
<th colspan="3" align="left" valign="top" rowspan="1"><hr></hr>
</th>
<th colspan="3" align="left" valign="top" rowspan="1"><hr></hr>
</th>
<th colspan="3" align="left" valign="top" rowspan="1"><hr></hr>
</th>
</tr>
<tr><th align="left" valign="top" rowspan="1" colspan="1"></th>
<th align="left" valign="top" rowspan="1" colspan="1">D3</th>
<th align="left" valign="top" rowspan="1" colspan="1">D6</th>
<th align="left" valign="top" rowspan="1" colspan="1">D9</th>
<th align="left" valign="top" rowspan="1" colspan="1">D3</th>
<th align="left" valign="top" rowspan="1" colspan="1">D6</th>
<th align="left" valign="top" rowspan="1" colspan="1">D9</th>
<th align="left" valign="top" rowspan="1" colspan="1">D3</th>
<th align="left" valign="top" rowspan="1" colspan="1">D6</th>
<th align="left" valign="top" rowspan="1" colspan="1">D9</th>
<th align="left" valign="top" rowspan="1" colspan="1">D3</th>
<th align="left" valign="top" rowspan="1" colspan="1">D6</th>
<th align="left" valign="top" rowspan="1" colspan="1">D9</th>
</tr>
</thead>
<tbody><tr><td align="left" valign="top" rowspan="1" colspan="1">NT</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
<td align="left" valign="top" rowspan="1" colspan="1">/</td>
<td align="left" valign="top" rowspan="1" colspan="1">/</td>
<td align="left" valign="top" rowspan="1" colspan="1">/</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
<td align="left" valign="top" rowspan="1" colspan="1">+2</td>
<td align="left" valign="top" rowspan="1" colspan="1">/</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">NT-F-L</td>
<td align="left" valign="top" rowspan="1" colspan="1">+2</td>
<td align="left" valign="top" rowspan="1" colspan="1">+2</td>
<td align="left" valign="top" rowspan="1" colspan="1">+2</td>
<td align="left" valign="top" rowspan="1" colspan="1">+3</td>
<td align="left" valign="top" rowspan="1" colspan="1">+2</td>
<td align="left" valign="top" rowspan="1" colspan="1">+2</td>
<td align="left" valign="top" rowspan="1" colspan="1">/</td>
<td align="left" valign="top" rowspan="1" colspan="1">/</td>
<td align="left" valign="top" rowspan="1" colspan="1">/</td>
<td align="left" valign="top" rowspan="1" colspan="1">+2</td>
<td align="left" valign="top" rowspan="1" colspan="1">+3</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">NT-F-M</td>
<td align="left" valign="top" rowspan="1" colspan="1">+3</td>
<td align="left" valign="top" rowspan="1" colspan="1">+2</td>
<td align="left" valign="top" rowspan="1" colspan="1">+4</td>
<td align="left" valign="top" rowspan="1" colspan="1">+4</td>
<td align="left" valign="top" rowspan="1" colspan="1">+4</td>
<td align="left" valign="top" rowspan="1" colspan="1">+2</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
<td align="left" valign="top" rowspan="1" colspan="1">+2</td>
<td align="left" valign="top" rowspan="1" colspan="1">+3</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">NT-F-H</td>
<td align="left" valign="top" rowspan="1" colspan="1">+4</td>
<td align="left" valign="top" rowspan="1" colspan="1">+3</td>
<td align="left" valign="top" rowspan="1" colspan="1">+3</td>
<td align="left" valign="top" rowspan="1" colspan="1">+2</td>
<td align="left" valign="top" rowspan="1" colspan="1">+3</td>
<td align="left" valign="top" rowspan="1" colspan="1">+2</td>
<td align="left" valign="top" rowspan="1" colspan="1">−</td>
<td align="left" valign="top" rowspan="1" colspan="1">+2</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
<td align="left" valign="top" rowspan="1" colspan="1">−</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
<td align="left" valign="top" rowspan="1" colspan="1">+</td>
</tr>
</tbody>
</table>
<table-wrap-foot><fn id="tfn8-ijn-7-1965"><p><bold>Notes: +</bold>
, Upregulated compared with PT (<italic>P</italic>
< 0.01); −, downregulated (<italic>P</italic>
< 0.01);<bold>/</bold>
, no significant difference compared with PT.</p>
</fn>
<fn id="tfn9-ijn-7-1965"><p><bold>Abbreviations:</bold>
<italic>ICAM1</italic>
, intercellular adhesion molecule 1; <italic>ITGB</italic>
, β-integrin; <italic>LAMA1</italic>
, laminin-1; NT, nanotube; PT, polished titanium; <italic>VEGFA,</italic>
vascular endothelial growth factor A.</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="Ma, Qianli" sort="Ma, Qianli" uniqKey="Ma Q" first="Qianli" last="Ma">Qianli Ma</name>
</noRegion>
<name sortKey="Chu, Paul K" sort="Chu, Paul K" uniqKey="Chu P" first="Paul K" last="Chu">Paul K. Chu</name>
<name sortKey="Ji, Kun" sort="Ji, Kun" uniqKey="Ji K" first="Kun" last="Ji">Kun Ji</name>
<name sortKey="Jin, Lei" sort="Jin, Lei" uniqKey="Jin L" first="Lei" last="Jin">Lei Jin</name>
<name sortKey="Mei, Shenglin" sort="Mei, Shenglin" uniqKey="Mei S" first="Shenglin" last="Mei">Shenglin Mei</name>
<name sortKey="Mei, Shenglin" sort="Mei, Shenglin" uniqKey="Mei S" first="Shenglin" last="Mei">Shenglin Mei</name>
<name sortKey="Wang, Wei" sort="Wang, Wei" uniqKey="Wang W" first="Wei" last="Wang">Wei Wang</name>
<name sortKey="Zhang, Yumei" sort="Zhang, Yumei" uniqKey="Zhang Y" first="Yumei" last="Zhang">Yumei Zhang</name>
</country>
</tree>
</affiliations>
</record>
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