Controlled release of nerve growth factor from heparin-conjugated fibrin gel within the nerve growth factor-delivering implant
Identifieur interne : 000A83 ( Ncbi/Merge ); précédent : 000A82; suivant : 000A84Controlled release of nerve growth factor from heparin-conjugated fibrin gel within the nerve growth factor-delivering implant
Auteurs : Jin-Yong Lee [Corée du Sud] ; Soung-Min Kim [Corée du Sud] ; Myung-Jin Kim [Corée du Sud] ; Jong-Ho Lee [Corée du Sud]Source :
- Journal of the Korean Association of Oral and Maxillofacial Surgeons [ 2234-7550 ] ; 2014.
Abstract
Although nerve growth factor (NGF) could promote the functional regeneration of an injured peripheral nerve, it is very difficult for NGF to sustain the therapeutic dose in the defect due to its short half-life. In this study, we loaded the NGF-bound heparin-conjugated fibrin (HCF) gel in the NGF-delivering implants and analyzed the time-dependent release of NGF and its bioactivity to evaluate the clinical effectiveness.
NGF solution was made of 1.0 mg of NGF and 1.0 mL of phosphate buffered saline (PBS). Experimental group A consisted of three implants, in which 0.25 µL of NGF solution, 0.75 µL of HCF, 1.0 µL of fibrinogen and 2.0 µL of thrombin was injected via apex hole with micropipette and gelated, were put into the centrifuge tube. Three implants of experimental group B were prepared with the mixture of 0.5 µL of NGF solution, 0.5 µL HCF, 1.0 µL of fibrinogen and 2.0 µL of thrombin. These six centrifuge tubes were filled with 1.0 mL of PBS and stirred in the water-filled beaker at 50 rpm. At 1, 3, 5, 7, 10, and 14 days, 1.0 mL of solution in each tubes was collected and preserved at -20℃ with adding same amount of fresh PBS. Enzyme-linked immunosorbent assay (ELISA) was done to determine
The average concentration of released NGF in the group A and B increased for the first 5 days and then gradually decreased. Almost all of NGF was released during 10 days. Released NGF from two groups could promote neural differentiation and neurite outgrowth of PC12 cells and these bioactivity was maintained over 14 days.
Controlled release system using NGF-HCF gel via NGF-delivering implant could be an another vehicle of delivering NGF to promote the nerve regeneration of dental implant related nerve damage.
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DOI: 10.5125/jkaoms.2014.40.1.3
PubMed: 24627836
PubMed Central: 3949490
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Controlled release of nerve growth factor from heparin-conjugated fibrin gel within the nerve growth factor-delivering implant</title><author><name sortKey="Lee, Jin Yong" sort="Lee, Jin Yong" uniqKey="Lee J" first="Jin-Yong" last="Lee">Jin-Yong Lee</name><affiliation wicri:level="3"><nlm:aff id="A1">Department of Oral and Maxillofacial Surgery, Korea University Guro Hospital, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Oral and Maxillofacial Surgery, Korea University Guro Hospital, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region></placeName></affiliation></author><author><name sortKey="Kim, Soung Min" sort="Kim, Soung Min" uniqKey="Kim S" first="Soung-Min" last="Kim">Soung-Min Kim</name><affiliation wicri:level="4"><nlm:aff id="A2">Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region><settlement type="city">Séoul</settlement></placeName><orgName type="university">Université nationale de Séoul</orgName></affiliation></author><author><name sortKey="Kim, Myung Jin" sort="Kim, Myung Jin" uniqKey="Kim M" first="Myung-Jin" last="Kim">Myung-Jin Kim</name><affiliation wicri:level="4"><nlm:aff id="A2">Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region><settlement type="city">Séoul</settlement></placeName><orgName type="university">Université nationale de Séoul</orgName></affiliation></author><author><name sortKey="Lee, Jong Ho" sort="Lee, Jong Ho" uniqKey="Lee J" first="Jong-Ho" last="Lee">Jong-Ho Lee</name><affiliation wicri:level="4"><nlm:aff id="A2">Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region><settlement type="city">Séoul</settlement></placeName><orgName type="university">Université nationale de 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within the nerve growth factor-delivering implant</title><author><name sortKey="Lee, Jin Yong" sort="Lee, Jin Yong" uniqKey="Lee J" first="Jin-Yong" last="Lee">Jin-Yong Lee</name><affiliation wicri:level="3"><nlm:aff id="A1">Department of Oral and Maxillofacial Surgery, Korea University Guro Hospital, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Oral and Maxillofacial Surgery, Korea University Guro Hospital, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region></placeName></affiliation></author><author><name sortKey="Kim, Soung Min" sort="Kim, Soung Min" uniqKey="Kim S" first="Soung-Min" last="Kim">Soung-Min Kim</name><affiliation wicri:level="4"><nlm:aff id="A2">Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region><settlement type="city">Séoul</settlement></placeName><orgName type="university">Université nationale de Séoul</orgName></affiliation></author><author><name sortKey="Kim, Myung Jin" sort="Kim, Myung Jin" uniqKey="Kim M" first="Myung-Jin" last="Kim">Myung-Jin Kim</name><affiliation wicri:level="4"><nlm:aff id="A2">Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region><settlement type="city">Séoul</settlement></placeName><orgName type="university">Université nationale de Séoul</orgName></affiliation></author><author><name sortKey="Lee, Jong Ho" sort="Lee, Jong Ho" uniqKey="Lee J" first="Jong-Ho" last="Lee">Jong-Ho Lee</name><affiliation wicri:level="4"><nlm:aff id="A2">Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region><settlement type="city">Séoul</settlement></placeName><orgName type="university">Université nationale de Séoul</orgName></affiliation></author></analytic><series><title level="j">Journal of the Korean Association of Oral and Maxillofacial Surgeons</title><idno type="ISSN">2234-7550</idno><idno type="eISSN">2234-5930</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Objectives</title><p>Although nerve growth factor (NGF) could promote the functional regeneration of an injured peripheral nerve, it is very difficult for NGF to sustain the therapeutic dose in the defect due to its short half-life. In this study, we loaded the NGF-bound heparin-conjugated fibrin (HCF) gel in the NGF-delivering implants and analyzed the time-dependent release of NGF and its bioactivity to evaluate the clinical effectiveness.</p></sec><sec><title>Materials and Methods</title><p>NGF solution was made of 1.0 mg of NGF and 1.0 mL of phosphate buffered saline (PBS). Experimental group A consisted of three implants, in which 0.25 µL of NGF solution, 0.75 µL of HCF, 1.0 µL of fibrinogen and 2.0 µL of thrombin was injected via apex hole with micropipette and gelated, were put into the centrifuge tube. Three implants of experimental group B were prepared with the mixture of 0.5 µL of NGF solution, 0.5 µL HCF, 1.0 µL of fibrinogen and 2.0 µL of thrombin. These six centrifuge tubes were filled with 1.0 mL of PBS and stirred in the water-filled beaker at 50 rpm. At 1, 3, 5, 7, 10, and 14 days, 1.0 mL of solution in each tubes was collected and preserved at -20℃ with adding same amount of fresh PBS. Enzyme-linked immunosorbent assay (ELISA) was done to determine <italic>in vitro</italic> release profile of NGF and its bioactivity was evaluated with neural differentiation of pheochromocytoma (PC12) cells.</p></sec><sec><title>Results</title><p>The average concentration of released NGF in the group A and B increased for the first 5 days and then gradually decreased. Almost all of NGF was released during 10 days. Released NGF from two groups could promote neural differentiation and neurite outgrowth of PC12 cells and these bioactivity was maintained over 14 days.</p></sec><sec><title>Conclusion</title><p>Controlled release system using NGF-HCF gel via NGF-delivering implant could be an another vehicle of delivering NGF to promote the nerve regeneration of dental implant related nerve damage.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Renton, T" uniqKey="Renton T">T Renton</name></author><author><name sortKey="Yilmaz, Z" uniqKey="Yilmaz Z">Z Yilmaz</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Van Steenberghe, D" uniqKey="Van Steenberghe D">D van Steenberghe</name></author><author><name sortKey="Lekholm, U" uniqKey="Lekholm U">U Lekholm</name></author><author><name sortKey="Bolender, C" uniqKey="Bolender C">C Bolender</name></author><author><name sortKey="Folmer, T" uniqKey="Folmer T">T 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<front><journal-meta><journal-id journal-id-type="nlm-ta">J Korean Assoc Oral Maxillofac Surg</journal-id><journal-id journal-id-type="iso-abbrev">J Korean Assoc Oral Maxillofac Surg</journal-id><journal-id journal-id-type="publisher-id">JKAOMS</journal-id><journal-title-group><journal-title>Journal of the Korean Association of Oral and Maxillofacial Surgeons</journal-title></journal-title-group><issn pub-type="ppub">2234-7550</issn><issn pub-type="epub">2234-5930</issn><publisher><publisher-name>The Korean Association of Oral and Maxillofacial Surgeons</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">24627836</article-id><article-id pub-id-type="pmc">3949490</article-id><article-id pub-id-type="doi">10.5125/jkaoms.2014.40.1.3</article-id><article-categories><subj-group><subject>Original Article</subject></subj-group></article-categories><title-group><article-title>Controlled release of nerve growth factor from heparin-conjugated fibrin gel within the nerve growth factor-delivering implant</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Lee</surname><given-names>Jin-Yong</given-names></name><xref ref-type="aff" rid="A1">1</xref></contrib><contrib contrib-type="author"><name><surname>Kim</surname><given-names>Soung-Min</given-names></name><xref ref-type="aff" rid="A2">2</xref></contrib><contrib contrib-type="author"><name><surname>Kim</surname><given-names>Myung-Jin</given-names></name><xref ref-type="aff" rid="A2">2</xref></contrib><contrib contrib-type="author" corresp="yes"><name><surname>Lee</surname><given-names>Jong-Ho</given-names></name><xref ref-type="aff" rid="A2">2</xref></contrib></contrib-group><aff id="A1"><label>1</label>Department of Oral and Maxillofacial Surgery, Korea University Guro Hospital, Seoul, Korea.</aff><aff id="A2"><label>2</label>Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul, Korea.</aff><author-notes><corresp>Corresponding author: Jong-Ho Lee. Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, 101, Daehak-ro, Jongno-gu, Seoul 110-768, Korea. TEL: +82-2-2072-2630, FAX: +82-2-766-4948, <email>leejongh@snu.ac.kr</email></corresp></author-notes><pub-date pub-type="ppub"><month>2</month><year>2014</year></pub-date><pub-date pub-type="epub"><day>25</day><month>2</month><year>2014</year></pub-date><volume>40</volume><issue>1</issue><fpage>3</fpage><lpage>10</lpage><history><date date-type="received"><day>23</day><month>10</month><year>2013</year></date><date date-type="rev-recd"><day>24</day><month>10</month><year>2013</year></date><date date-type="accepted"><day>25</day><month>10</month><year>2013</year></date></history><permissions><copyright-statement>Copyright © 2014 The Korean Association of Oral and Maxillofacial Surgeons. All rights reserved.</copyright-statement><copyright-year>2014</copyright-year><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by-nc/3.0/"><license-p>This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (<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>) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p></license></permissions><abstract><sec><title>Objectives</title><p>Although nerve growth factor (NGF) could promote the functional regeneration of an injured peripheral nerve, it is very difficult for NGF to sustain the therapeutic dose in the defect due to its short half-life. In this study, we loaded the NGF-bound heparin-conjugated fibrin (HCF) gel in the NGF-delivering implants and analyzed the time-dependent release of NGF and its bioactivity to evaluate the clinical effectiveness.</p></sec><sec><title>Materials and Methods</title><p>NGF solution was made of 1.0 mg of NGF and 1.0 mL of phosphate buffered saline (PBS). Experimental group A consisted of three implants, in which 0.25 µL of NGF solution, 0.75 µL of HCF, 1.0 µL of fibrinogen and 2.0 µL of thrombin was injected via apex hole with micropipette and gelated, were put into the centrifuge tube. Three implants of experimental group B were prepared with the mixture of 0.5 µL of NGF solution, 0.5 µL HCF, 1.0 µL of fibrinogen and 2.0 µL of thrombin. These six centrifuge tubes were filled with 1.0 mL of PBS and stirred in the water-filled beaker at 50 rpm. At 1, 3, 5, 7, 10, and 14 days, 1.0 mL of solution in each tubes was collected and preserved at -20℃ with adding same amount of fresh PBS. Enzyme-linked immunosorbent assay (ELISA) was done to determine <italic>in vitro</italic> release profile of NGF and its bioactivity was evaluated with neural differentiation of pheochromocytoma (PC12) cells.</p></sec><sec><title>Results</title><p>The average concentration of released NGF in the group A and B increased for the first 5 days and then gradually decreased. Almost all of NGF was released during 10 days. Released NGF from two groups could promote neural differentiation and neurite outgrowth of PC12 cells and these bioactivity was maintained over 14 days.</p></sec><sec><title>Conclusion</title><p>Controlled release system using NGF-HCF gel via NGF-delivering implant could be an another vehicle of delivering NGF to promote the nerve regeneration of dental implant related nerve damage.</p></sec></abstract><kwd-group><kwd>Nerve growth factor-delivering implant</kwd><kwd>Nerve growth factor</kwd><kwd>Controlled release</kwd><kwd>Nerve regeneration</kwd></kwd-group><funding-group><award-group><funding-source country="KR">The Korea Healthcare Technology R&D Project, Ministry for Health, Welfare & Family Affairs, Republic of Korea</funding-source><award-id>A101578</award-id></award-group></funding-group></article-meta></front><floats-group><fig id="F1" orientation="portrait" position="float"><label>Fig. 1</label><caption><p>Design of nerve growth factor (NGF) delivering implant. A. Schematic design. B. Photographic images of NGF delivering implant.</p></caption><graphic xlink:href="jkaoms-40-3-g001"></graphic></fig><fig id="F2" orientation="portrait" position="float"><label>Fig. 2</label><caption><p><italic>In vitro</italic> profile of NGF release from NGF-HCF gel loaded in NGF delivering implant at each time point of sampling (pg/mL). The amount of released NGF of two groups were increased and reached peak at days 5 and decreased. At day 3, released NGF of group A (459.666±302.739 pg/mL) was higher than that of group B (66.856±42.352 pg/mL), but there was no significance (Mann-Whitney U-test). (NGF: nerve growth factor, HCF: heparin-conjugated fibrin)</p></caption><graphic xlink:href="jkaoms-40-3-g002"></graphic></fig><fig id="F3" orientation="portrait" position="float"><label>Fig. 3</label><caption><p>Cumulative profile of released NGF from NGF-HCF gel loaded in NGF delivering implant (%). The cumulative release profile of NGF showed a similar pattern between two experimental groups over 14 days. For 10 days, almost all of NGF from NGF-HCF gel loaded in the NGF delivering implant was released. (NGF: nerve growth factor, HCF: heparin-conjugated fibrin)</p></caption><graphic xlink:href="jkaoms-40-3-g003"></graphic></fig><fig id="F4" orientation="portrait" position="float"><label>Fig. 4</label><caption><p>Microscopic images of the differentiated PC12 cells (×100). A. PC12 cells cultured with medium containing group A sample harvested at day 5. B. PC12 cells cultured with medium containing group B sample harvested at day 5. C. PC12 cells cultured with medium without NGF for 3 days (control group 1). D. PC12 cells cultured with medium without nerve growth factor for 6 days (control group 2). Scale bars=50 µm.</p></caption><graphic xlink:href="jkaoms-40-3-g004"></graphic></fig><fig id="F5" orientation="portrait" position="float"><label>Fig. 5</label><caption><p>Comparison of the ratio of differentiated PC12 cells in group A. A. Comparison with control group 1. The differentiation ratio at day 3 and 5 were significantly higher than that of control group 1. B. Comparison with control group 2. The ratio days 1, 3, 5, and 7 were higher than that of control group 2 with significance (<sup>*</sup><italic>P</italic><0.05, <sup>***</sup><italic>P</italic><0.001; one-way ANOVA with Bonferroni's post hoc test). (NGF: nerve growth factor)</p></caption><graphic xlink:href="jkaoms-40-3-g005"></graphic></fig><fig id="F6" orientation="portrait" position="float"><label>Fig. 6</label><caption><p>Comparison of the ratio of differentiated PC12 cells in group B. A. Comparison with control group 1. B. Comparison with control group 2. The differentiation ratio of PC12 cells at days 3, 5, and 7 were significantly higher than those of control group 1 and 2 (<sup>*</sup><italic>P</italic><0.05, <sup>**</sup><italic>P</italic><0.01, <sup>***</sup><italic>P</italic><0.001; one-way ANOVA with Bonferroni's post hoc test). (NGF: nerve growth factor)</p></caption><graphic xlink:href="jkaoms-40-3-g006"></graphic></fig><fig id="F7" orientation="portrait" position="float"><label>Fig. 7</label><caption><p>Comparison of the neurite length of differentiated PC12 cells in group A. A. Comparison with control group 1. B. Comparison with control group 2. No significant differences were found between group A, control group 1 and 2 (one-way ANOVA with Bonferroni's post hoc test). (NGF: nerve growth factor)</p></caption><graphic xlink:href="jkaoms-40-3-g007"></graphic></fig><fig id="F8" orientation="portrait" position="float"><label>Fig. 8</label><caption><p>Comparison of the neurite length of differentiated PC12 cells in group B. A. Comparison with control group 1. B. Comparison with control group 2. The average length of neurite of day 1 was significantly higher than those of control group 1 and 2 (<sup>*</sup><italic>P</italic><0.05; one-way ANOVA with Bonferroni's post hoc test). (NGF: nerve growth factor)</p></caption><graphic xlink:href="jkaoms-40-3-g008"></graphic></fig><table-wrap id="T1" orientation="portrait" position="float"><label>Table 1</label><caption><p>Composition of NGF-HCF gel</p></caption><graphic xlink:href="jkaoms-40-3-i001"></graphic><table-wrap-foot><fn><p>(NGF: nerve growth factor, HCF: heparin-conjugated fibrin)</p></fn></table-wrap-foot></table-wrap></floats-group></pmc><affiliations><list><country><li>Corée du Sud</li></country><region><li>Région capitale de Séoul</li></region><settlement><li>Séoul</li></settlement><orgName><li>Université nationale de Séoul</li></orgName></list><tree><country name="Corée du Sud"><region name="Région capitale de Séoul"><name sortKey="Lee, Jin Yong" sort="Lee, Jin Yong" uniqKey="Lee J" first="Jin-Yong" last="Lee">Jin-Yong Lee</name></region><name sortKey="Kim, Myung Jin" sort="Kim, Myung Jin" uniqKey="Kim M" first="Myung-Jin" last="Kim">Myung-Jin Kim</name><name sortKey="Kim, Soung Min" sort="Kim, Soung Min" uniqKey="Kim S" first="Soung-Min" last="Kim">Soung-Min Kim</name><name sortKey="Lee, Jong Ho" sort="Lee, Jong Ho" uniqKey="Lee J" first="Jong-Ho" last="Lee">Jong-Ho Lee</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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