Adenovirus-mediated GDF-5 promotes the extracellular matrix expression in degenerative nucleus pulposus cells
Objective To construct a recombinant adenovirus vector-carrying human growth and differentiation factor-5 (GDF-5) gene, investigate the biological effects of adenovirus-mediated GDF-5 (Ad-GDF-5) on extracellular matrix (ECM) expression in human degenerative disc nucleus pulposus (NP) cells, and expl...
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| Veröffentlicht in: | Journal of Zhejiang University. B. Science 2016, Vol.17 (1), p.30-42 |
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| creator | Luo, Xu-wei Liu, Kang Chen, Zhu Zhao, Ming Han, Xiao-wei Bai, Yi-guang Feng, Gang |
| description | Objective
To construct a recombinant adenovirus vector-carrying human growth and differentiation factor-5 (GDF-5) gene, investigate the biological effects of adenovirus-mediated GDF-5 (Ad-GDF-5) on extracellular matrix (ECM) expression in human degenerative disc nucleus pulposus (NP) cells, and explore a candidate gene therapy method for intervertebral disc degeneration (IDD).
Methods
Human NP cells of a degenerative disc were isolated, cultured, and infected with Ad-GDF-5 using the AdEasy-1 adenovirus vector system. On Days 3, 7, 14, and 21, the contents of the sulfated glycosaminoglycan (sGAG), deoxyribonucleic acid (DNA) and hydroxyproline (Hyp), synthesis of proteoglycan and collagen II, gene expression of collagen II and aggrecan, and NP cell proliferation were assessed.
Results
The adenovirus was an effective vehicle for gene delivery with prolonged expression of GDF-5. Biochemical analysis revealed increased sGAG and Hyp contents in human NP cells infected by Ad-GDF-5 whereas there was no conspicuous change in basal medium (BM) or Ad-green fluorescent protein (GFP) groups. Only cells in the Ad-GDF-5 group promoted the production of ECM, as demonstrated by the secretion of proteoglycan and up-regulation of collagen II and aggrecan at both protein and mRNA levels. The NP cell proliferation was significantly promoted.
Conclusions
The data suggest that Ad-GDF-5 gene therapy is a potential treatment for IDD, which restores the functions of degenerative intervertebral disc through enhancing the ECM production of human NP cells. |
| doi_str_mv | 10.1631/jzus.B1500182 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4710838</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1825431075</sourcerecordid><originalsourceid>FETCH-LOGICAL-c564t-71c60515eefb66f884622173334f0fad0e765f0ea3640a5a2cc514ebdf619c363</originalsourceid><addsrcrecordid>eNqNkktv1DAUhSNERR-wZIsisekmg2_8zAaplLYgVWIDa8uT3Ew9Suxgx6PSX4-jaauCkOjKls_nY5-rUxRvgaxAUPiwvUtx9Qk4IaDqF8URKFFXIBV9mfdC0gq4gsPiOMYtIYwRKV4Vh3UWGl6zo2I869D5nQ0pViN21szYlVefLyteTsGPfsZYzjdY4u0cTIvDkAYTytHMwd7mwylgjNa70rqyww06DGa2OyxdagdMsZzSMPmYN8vd-Lo46M0Q8c39elL8uLz4fv6luv529fX87LpquWBzJaEVhANH7NdC9EoxUdcgKaWsJ73pCErBe4KGCkYMN3XbcmC47noBTUsFPSk-7n2ntM6pWnT594Oegh1N-KW9sfpPxdkbvfE7zSQQRVU2OL03CP5nwjjr0cYlgnHoU9R51pxRIJL_H5UyG9aqbp6BCqIayhrI6Pu_0K1PweWhaciyErKRS85qT7XBxxiwf4wIRC_10Es99EM9Mv_u6Vwe6Yc-ZGC1B2KW3AbDk2f_6fgb9qzHTw</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1913867976</pqid></control><display><type>article</type><title>Adenovirus-mediated GDF-5 promotes the extracellular matrix expression in degenerative nucleus pulposus cells</title><source>MEDLINE</source><source>SpringerLink Journals</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>PubMed Central</source><source>Free Full-Text Journals in Chemistry</source><creator>Luo, Xu-wei ; Liu, Kang ; Chen, Zhu ; Zhao, Ming ; Han, Xiao-wei ; Bai, Yi-guang ; Feng, Gang</creator><creatorcontrib>Luo, Xu-wei ; Liu, Kang ; Chen, Zhu ; Zhao, Ming ; Han, Xiao-wei ; Bai, Yi-guang ; Feng, Gang</creatorcontrib><description>Objective
To construct a recombinant adenovirus vector-carrying human growth and differentiation factor-5 (GDF-5) gene, investigate the biological effects of adenovirus-mediated GDF-5 (Ad-GDF-5) on extracellular matrix (ECM) expression in human degenerative disc nucleus pulposus (NP) cells, and explore a candidate gene therapy method for intervertebral disc degeneration (IDD).
Methods
Human NP cells of a degenerative disc were isolated, cultured, and infected with Ad-GDF-5 using the AdEasy-1 adenovirus vector system. On Days 3, 7, 14, and 21, the contents of the sulfated glycosaminoglycan (sGAG), deoxyribonucleic acid (DNA) and hydroxyproline (Hyp), synthesis of proteoglycan and collagen II, gene expression of collagen II and aggrecan, and NP cell proliferation were assessed.
Results
The adenovirus was an effective vehicle for gene delivery with prolonged expression of GDF-5. Biochemical analysis revealed increased sGAG and Hyp contents in human NP cells infected by Ad-GDF-5 whereas there was no conspicuous change in basal medium (BM) or Ad-green fluorescent protein (GFP) groups. Only cells in the Ad-GDF-5 group promoted the production of ECM, as demonstrated by the secretion of proteoglycan and up-regulation of collagen II and aggrecan at both protein and mRNA levels. The NP cell proliferation was significantly promoted.
Conclusions
The data suggest that Ad-GDF-5 gene therapy is a potential treatment for IDD, which restores the functions of degenerative intervertebral disc through enhancing the ECM production of human NP cells.</description><identifier>ISSN: 1673-1581</identifier><identifier>EISSN: 1862-1783</identifier><identifier>DOI: 10.1631/jzus.B1500182</identifier><identifier>PMID: 26739524</identifier><language>eng</language><publisher>Hangzhou: Zhejiang University Press</publisher><subject>Adenoviridae - physiology ; Adenovirus ; Adenoviruses ; Aggrecan ; bioactive properties ; Biochemical analysis ; Biological effects ; Biomedical and Life Sciences ; Biomedicine ; Cell growth ; Cell Line ; Cell proliferation ; China ; Collagen ; Collagen (type II) ; culture media ; Degeneration ; Deoxyribonucleic acid ; Differentiation ; DNA ; DNA biosynthesis ; Extracellular matrix ; Extracellular Matrix - metabolism ; Extracellular Matrix - ultrastructure ; Extracellular Matrix Proteins - metabolism ; Fluorescence ; fluorescent proteins ; Gene expression ; gene expression regulation ; Gene therapy ; Gene transfer ; genes ; glycosaminoglycans ; Green fluorescent protein ; Growth differentiation factor 5 ; Growth Differentiation Factor 5 - genetics ; Growth Differentiation Factor 5 - metabolism ; human growth ; Humans ; Hydroxyproline ; Intervertebral Disc - growth & development ; Intervertebral Disc - ultrastructure ; Intervertebral Disc Degeneration - metabolism ; Intervertebral Disc Degeneration - pathology ; Intervertebral discs ; intervertebral disks ; messenger RNA ; Nuclei ; Nuclei (cytology) ; Nucleus pulposus ; proteoglycans ; Secretion ; Therapeutic applications ; Transduction, Genetic - methods</subject><ispartof>Journal of Zhejiang University. B. Science, 2016, Vol.17 (1), p.30-42</ispartof><rights>Zhejiang University and Springer-Verlag Berlin Heidelberg 2016</rights><rights>Journal of Zhejiang University SCIENCE B is a copyright of Springer, 2016.</rights><rights>Copyright © Zhejiang University and Springer-Verlag Berlin Heidelberg 2016 2016</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c564t-71c60515eefb66f884622173334f0fad0e765f0ea3640a5a2cc514ebdf619c363</citedby><cites>FETCH-LOGICAL-c564t-71c60515eefb66f884622173334f0fad0e765f0ea3640a5a2cc514ebdf619c363</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4710838/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4710838/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,724,777,781,882,27905,27906,41469,42538,51300,53772,53774</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26739524$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Luo, Xu-wei</creatorcontrib><creatorcontrib>Liu, Kang</creatorcontrib><creatorcontrib>Chen, Zhu</creatorcontrib><creatorcontrib>Zhao, Ming</creatorcontrib><creatorcontrib>Han, Xiao-wei</creatorcontrib><creatorcontrib>Bai, Yi-guang</creatorcontrib><creatorcontrib>Feng, Gang</creatorcontrib><title>Adenovirus-mediated GDF-5 promotes the extracellular matrix expression in degenerative nucleus pulposus cells</title><title>Journal of Zhejiang University. B. Science</title><addtitle>J. Zhejiang Univ. Sci. B</addtitle><addtitle>J Zhejiang Univ Sci B</addtitle><description>Objective
To construct a recombinant adenovirus vector-carrying human growth and differentiation factor-5 (GDF-5) gene, investigate the biological effects of adenovirus-mediated GDF-5 (Ad-GDF-5) on extracellular matrix (ECM) expression in human degenerative disc nucleus pulposus (NP) cells, and explore a candidate gene therapy method for intervertebral disc degeneration (IDD).
Methods
Human NP cells of a degenerative disc were isolated, cultured, and infected with Ad-GDF-5 using the AdEasy-1 adenovirus vector system. On Days 3, 7, 14, and 21, the contents of the sulfated glycosaminoglycan (sGAG), deoxyribonucleic acid (DNA) and hydroxyproline (Hyp), synthesis of proteoglycan and collagen II, gene expression of collagen II and aggrecan, and NP cell proliferation were assessed.
Results
The adenovirus was an effective vehicle for gene delivery with prolonged expression of GDF-5. Biochemical analysis revealed increased sGAG and Hyp contents in human NP cells infected by Ad-GDF-5 whereas there was no conspicuous change in basal medium (BM) or Ad-green fluorescent protein (GFP) groups. Only cells in the Ad-GDF-5 group promoted the production of ECM, as demonstrated by the secretion of proteoglycan and up-regulation of collagen II and aggrecan at both protein and mRNA levels. The NP cell proliferation was significantly promoted.
Conclusions
The data suggest that Ad-GDF-5 gene therapy is a potential treatment for IDD, which restores the functions of degenerative intervertebral disc through enhancing the ECM production of human NP cells.</description><subject>Adenoviridae - physiology</subject><subject>Adenovirus</subject><subject>Adenoviruses</subject><subject>Aggrecan</subject><subject>bioactive properties</subject><subject>Biochemical analysis</subject><subject>Biological effects</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Cell growth</subject><subject>Cell Line</subject><subject>Cell proliferation</subject><subject>China</subject><subject>Collagen</subject><subject>Collagen (type II)</subject><subject>culture media</subject><subject>Degeneration</subject><subject>Deoxyribonucleic acid</subject><subject>Differentiation</subject><subject>DNA</subject><subject>DNA biosynthesis</subject><subject>Extracellular matrix</subject><subject>Extracellular Matrix - metabolism</subject><subject>Extracellular Matrix - ultrastructure</subject><subject>Extracellular Matrix Proteins - metabolism</subject><subject>Fluorescence</subject><subject>fluorescent proteins</subject><subject>Gene expression</subject><subject>gene expression regulation</subject><subject>Gene therapy</subject><subject>Gene transfer</subject><subject>genes</subject><subject>glycosaminoglycans</subject><subject>Green fluorescent protein</subject><subject>Growth differentiation factor 5</subject><subject>Growth Differentiation Factor 5 - genetics</subject><subject>Growth Differentiation Factor 5 - metabolism</subject><subject>human growth</subject><subject>Humans</subject><subject>Hydroxyproline</subject><subject>Intervertebral Disc - growth & development</subject><subject>Intervertebral Disc - ultrastructure</subject><subject>Intervertebral Disc Degeneration - metabolism</subject><subject>Intervertebral Disc Degeneration - pathology</subject><subject>Intervertebral discs</subject><subject>intervertebral disks</subject><subject>messenger RNA</subject><subject>Nuclei</subject><subject>Nuclei (cytology)</subject><subject>Nucleus pulposus</subject><subject>proteoglycans</subject><subject>Secretion</subject><subject>Therapeutic applications</subject><subject>Transduction, Genetic - methods</subject><issn>1673-1581</issn><issn>1862-1783</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqNkktv1DAUhSNERR-wZIsisekmg2_8zAaplLYgVWIDa8uT3Ew9Suxgx6PSX4-jaauCkOjKls_nY5-rUxRvgaxAUPiwvUtx9Qk4IaDqF8URKFFXIBV9mfdC0gq4gsPiOMYtIYwRKV4Vh3UWGl6zo2I869D5nQ0pViN21szYlVefLyteTsGPfsZYzjdY4u0cTIvDkAYTytHMwd7mwylgjNa70rqyww06DGa2OyxdagdMsZzSMPmYN8vd-Lo46M0Q8c39elL8uLz4fv6luv529fX87LpquWBzJaEVhANH7NdC9EoxUdcgKaWsJ73pCErBe4KGCkYMN3XbcmC47noBTUsFPSk-7n2ntM6pWnT594Oegh1N-KW9sfpPxdkbvfE7zSQQRVU2OL03CP5nwjjr0cYlgnHoU9R51pxRIJL_H5UyG9aqbp6BCqIayhrI6Pu_0K1PweWhaciyErKRS85qT7XBxxiwf4wIRC_10Es99EM9Mv_u6Vwe6Yc-ZGC1B2KW3AbDk2f_6fgb9qzHTw</recordid><startdate>2016</startdate><enddate>2016</enddate><creator>Luo, Xu-wei</creator><creator>Liu, Kang</creator><creator>Chen, Zhu</creator><creator>Zhao, Ming</creator><creator>Han, Xiao-wei</creator><creator>Bai, Yi-guang</creator><creator>Feng, Gang</creator><general>Zhejiang University Press</general><general>Springer Nature B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QO</scope><scope>7QP</scope><scope>7TK</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>BKSAR</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>M7S</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PCBAR</scope><scope>PDBOC</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>7X8</scope><scope>7U9</scope><scope>H94</scope><scope>7S9</scope><scope>L.6</scope><scope>5PM</scope></search><sort><creationdate>2016</creationdate><title>Adenovirus-mediated GDF-5 promotes the extracellular matrix expression in degenerative nucleus pulposus cells</title><author>Luo, Xu-wei ; Liu, Kang ; Chen, Zhu ; Zhao, Ming ; Han, Xiao-wei ; Bai, Yi-guang ; Feng, Gang</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c564t-71c60515eefb66f884622173334f0fad0e765f0ea3640a5a2cc514ebdf619c363</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Adenoviridae - physiology</topic><topic>Adenovirus</topic><topic>Adenoviruses</topic><topic>Aggrecan</topic><topic>bioactive properties</topic><topic>Biochemical analysis</topic><topic>Biological effects</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedicine</topic><topic>Cell growth</topic><topic>Cell Line</topic><topic>Cell proliferation</topic><topic>China</topic><topic>Collagen</topic><topic>Collagen (type II)</topic><topic>culture media</topic><topic>Degeneration</topic><topic>Deoxyribonucleic acid</topic><topic>Differentiation</topic><topic>DNA</topic><topic>DNA biosynthesis</topic><topic>Extracellular matrix</topic><topic>Extracellular Matrix - metabolism</topic><topic>Extracellular Matrix - ultrastructure</topic><topic>Extracellular Matrix Proteins - metabolism</topic><topic>Fluorescence</topic><topic>fluorescent proteins</topic><topic>Gene expression</topic><topic>gene expression regulation</topic><topic>Gene therapy</topic><topic>Gene transfer</topic><topic>genes</topic><topic>glycosaminoglycans</topic><topic>Green fluorescent protein</topic><topic>Growth differentiation factor 5</topic><topic>Growth Differentiation Factor 5 - genetics</topic><topic>Growth Differentiation Factor 5 - metabolism</topic><topic>human growth</topic><topic>Humans</topic><topic>Hydroxyproline</topic><topic>Intervertebral Disc - growth & development</topic><topic>Intervertebral Disc - ultrastructure</topic><topic>Intervertebral Disc Degeneration - metabolism</topic><topic>Intervertebral Disc Degeneration - pathology</topic><topic>Intervertebral discs</topic><topic>intervertebral disks</topic><topic>messenger RNA</topic><topic>Nuclei</topic><topic>Nuclei (cytology)</topic><topic>Nucleus pulposus</topic><topic>proteoglycans</topic><topic>Secretion</topic><topic>Therapeutic applications</topic><topic>Transduction, Genetic - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Luo, Xu-wei</creatorcontrib><creatorcontrib>Liu, Kang</creatorcontrib><creatorcontrib>Chen, Zhu</creatorcontrib><creatorcontrib>Zhao, Ming</creatorcontrib><creatorcontrib>Han, Xiao-wei</creatorcontrib><creatorcontrib>Bai, Yi-guang</creatorcontrib><creatorcontrib>Feng, Gang</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Biotechnology Research Abstracts</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Earth, Atmospheric & Aquatic Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>ProQuest Engineering Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>Engineering Database</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>Earth, Atmospheric & Aquatic Science Database</collection><collection>Materials Science Collection</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>Engineering Collection</collection><collection>Environmental Science Collection</collection><collection>MEDLINE - Academic</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of Zhejiang University. B. Science</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Luo, Xu-wei</au><au>Liu, Kang</au><au>Chen, Zhu</au><au>Zhao, Ming</au><au>Han, Xiao-wei</au><au>Bai, Yi-guang</au><au>Feng, Gang</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Adenovirus-mediated GDF-5 promotes the extracellular matrix expression in degenerative nucleus pulposus cells</atitle><jtitle>Journal of Zhejiang University. B. Science</jtitle><stitle>J. Zhejiang Univ. Sci. B</stitle><addtitle>J Zhejiang Univ Sci B</addtitle><date>2016</date><risdate>2016</risdate><volume>17</volume><issue>1</issue><spage>30</spage><epage>42</epage><pages>30-42</pages><issn>1673-1581</issn><eissn>1862-1783</eissn><abstract>Objective
To construct a recombinant adenovirus vector-carrying human growth and differentiation factor-5 (GDF-5) gene, investigate the biological effects of adenovirus-mediated GDF-5 (Ad-GDF-5) on extracellular matrix (ECM) expression in human degenerative disc nucleus pulposus (NP) cells, and explore a candidate gene therapy method for intervertebral disc degeneration (IDD).
Methods
Human NP cells of a degenerative disc were isolated, cultured, and infected with Ad-GDF-5 using the AdEasy-1 adenovirus vector system. On Days 3, 7, 14, and 21, the contents of the sulfated glycosaminoglycan (sGAG), deoxyribonucleic acid (DNA) and hydroxyproline (Hyp), synthesis of proteoglycan and collagen II, gene expression of collagen II and aggrecan, and NP cell proliferation were assessed.
Results
The adenovirus was an effective vehicle for gene delivery with prolonged expression of GDF-5. Biochemical analysis revealed increased sGAG and Hyp contents in human NP cells infected by Ad-GDF-5 whereas there was no conspicuous change in basal medium (BM) or Ad-green fluorescent protein (GFP) groups. Only cells in the Ad-GDF-5 group promoted the production of ECM, as demonstrated by the secretion of proteoglycan and up-regulation of collagen II and aggrecan at both protein and mRNA levels. The NP cell proliferation was significantly promoted.
Conclusions
The data suggest that Ad-GDF-5 gene therapy is a potential treatment for IDD, which restores the functions of degenerative intervertebral disc through enhancing the ECM production of human NP cells.</abstract><cop>Hangzhou</cop><pub>Zhejiang University Press</pub><pmid>26739524</pmid><doi>10.1631/jzus.B1500182</doi><tpages>13</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Adenoviridae - physiology Adenovirus Adenoviruses Aggrecan bioactive properties Biochemical analysis Biological effects Biomedical and Life Sciences Biomedicine Cell growth Cell Line Cell proliferation China Collagen Collagen (type II) culture media Degeneration Deoxyribonucleic acid Differentiation DNA DNA biosynthesis Extracellular matrix Extracellular Matrix - metabolism Extracellular Matrix - ultrastructure Extracellular Matrix Proteins - metabolism Fluorescence fluorescent proteins Gene expression gene expression regulation Gene therapy Gene transfer genes glycosaminoglycans Green fluorescent protein Growth differentiation factor 5 Growth Differentiation Factor 5 - genetics Growth Differentiation Factor 5 - metabolism human growth Humans Hydroxyproline Intervertebral Disc - growth & development Intervertebral Disc - ultrastructure Intervertebral Disc Degeneration - metabolism Intervertebral Disc Degeneration - pathology Intervertebral discs intervertebral disks messenger RNA Nuclei Nuclei (cytology) Nucleus pulposus proteoglycans Secretion Therapeutic applications Transduction, Genetic - methods |
| title | Adenovirus-mediated GDF-5 promotes the extracellular matrix expression in degenerative nucleus pulposus cells |
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