Gene-expression analysis of cementoblasts and osteoblasts
Background and Objective Cementum and bone are similar mineralized tissues, but cementum accumulates much more slowly than bone, does not have vasculature or innervation and does not undergo remodeling. Despite these differences, there are no well‐established markers to distinguish cementoblasts fro...
Gespeichert in:
| Veröffentlicht in: | Journal of periodontal research 2016-06, Vol.51 (3), p.304-312 |
|---|---|
| Hauptverfasser: | , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 312 |
|---|---|
| container_issue | 3 |
| container_start_page | 304 |
| container_title | Journal of periodontal research |
| container_volume | 51 |
| creator | Matthews, B. G. Roguljic, H. Franceschetti, T. Roeder, E. Matic, I. Vidovic, I. Joshi, P. Kum, K.-Y. Kalajzic, I. |
| description | Background and Objective
Cementum and bone are similar mineralized tissues, but cementum accumulates much more slowly than bone, does not have vasculature or innervation and does not undergo remodeling. Despite these differences, there are no well‐established markers to distinguish cementoblasts from other mature mineralizing cells such as osteoblasts and odontoblasts. The purpose of this study was to assess differences in gene expression between cementoblasts and osteoblasts using gene profiling of cell populations isolated directly from osteocalcin–green fluorescent protein (OC‐GFP) transgenic mice.
Material and Methods
OC‐GFP reporter mice were used as they show labeling of cementoblasts, osteoblasts and odontoblasts, but not of periodontal ligament fibroblasts, within the periodontium. We sorted cells digested from the molar root surface to isolate OC‐GFP+ cementoblasts. Osteoblasts were isolated from calvarial digests. Microarray analysis was performed, and selected results were confirmed by real‐time PCR and immunostaining or in situ hybridization.
Results
Microarray analysis identified 95 genes that were expressed at least two‐fold higher in cementoblasts than in osteoblasts. Our analysis indicated that the Wnt signaling pathway was differentially regulated, as were genes related to skeletal development. Real‐time PCR confirmed that expression of the Wnt inhibitors Wnt inhibitory factor 1 (Wif1) and secreted frizzled‐related protein 1 (Sfrp1) was elevated in cementoblasts compared with osteoblasts, and Wif1 expression was localized to the apical root region. In addition, the transcription factor BARX homeobox 1 (Barx1) was expressed at higher levels in cementoblasts, and immunohistochemistry indicated that BARX1 was expressed in apical cementoblasts and cementocytes, but not in osteoblasts or odontoblasts.
Conclusion
The OC‐GFP mouse provides a good model for selectively isolating cementoblasts, and allowed for identification of differentially expressed genes between cementoblasts and osteoblasts. |
| doi_str_mv | 10.1111/jre.12309 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4729669</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1784750606</sourcerecordid><originalsourceid>FETCH-LOGICAL-c4869-78f3f9b9e9d25cf279c0af3968653b40f926e85f62fce5ae966e5ccfcbf330313</originalsourceid><addsrcrecordid>eNqFkUtLAzEQx4MoWh8Hv4D0qIfVPDbJ5iJo0fpCUXzdQjad6Op2U5NW7bc32lr0IOYShvnNjxn-CK0TvE3S23kKsE0ow2oOtYjAOMNS8HnUwpjSjOVFvoSWY3zCqRZSLaIlKijhjIgWUl1oIIP3QYAYK9-0TWPqcaxi27u2hT40Q1_WJg5j6vTaPg5hWq-iBWfqCGvTfwXdHB5cd46ys4vucWfvLLN5IVQmC8ecKhWoHuXWUaksNo4pUQjOyhw7RQUU3AnqLHADSgjg1jpbOsYwI2wF7U68g1HZh55NGwVT60Go-iaMtTeV_t1pqkf94F91LmmSqSTYnAqCfxlBHOp-FS3UtWnAj6ImBea5Ulzh_1FZ5JJjgUVCtyaoDT7GAG62EcH6MxWdUtFfqSR24-cJM_I7hgTsTIC3qobx3yZ9cnXwrcwmE1VK5H02YcKzFpJJru_Ou_rk-vK2c3q_rzn7AOc2ptU</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1784750606</pqid></control><display><type>article</type><title>Gene-expression analysis of cementoblasts and osteoblasts</title><source>MEDLINE</source><source>Wiley Online Library Journals Frontfile Complete</source><creator>Matthews, B. G. ; Roguljic, H. ; Franceschetti, T. ; Roeder, E. ; Matic, I. ; Vidovic, I. ; Joshi, P. ; Kum, K.-Y. ; Kalajzic, I.</creator><creatorcontrib>Matthews, B. G. ; Roguljic, H. ; Franceschetti, T. ; Roeder, E. ; Matic, I. ; Vidovic, I. ; Joshi, P. ; Kum, K.-Y. ; Kalajzic, I.</creatorcontrib><description>Background and Objective
Cementum and bone are similar mineralized tissues, but cementum accumulates much more slowly than bone, does not have vasculature or innervation and does not undergo remodeling. Despite these differences, there are no well‐established markers to distinguish cementoblasts from other mature mineralizing cells such as osteoblasts and odontoblasts. The purpose of this study was to assess differences in gene expression between cementoblasts and osteoblasts using gene profiling of cell populations isolated directly from osteocalcin–green fluorescent protein (OC‐GFP) transgenic mice.
Material and Methods
OC‐GFP reporter mice were used as they show labeling of cementoblasts, osteoblasts and odontoblasts, but not of periodontal ligament fibroblasts, within the periodontium. We sorted cells digested from the molar root surface to isolate OC‐GFP+ cementoblasts. Osteoblasts were isolated from calvarial digests. Microarray analysis was performed, and selected results were confirmed by real‐time PCR and immunostaining or in situ hybridization.
Results
Microarray analysis identified 95 genes that were expressed at least two‐fold higher in cementoblasts than in osteoblasts. Our analysis indicated that the Wnt signaling pathway was differentially regulated, as were genes related to skeletal development. Real‐time PCR confirmed that expression of the Wnt inhibitors Wnt inhibitory factor 1 (Wif1) and secreted frizzled‐related protein 1 (Sfrp1) was elevated in cementoblasts compared with osteoblasts, and Wif1 expression was localized to the apical root region. In addition, the transcription factor BARX homeobox 1 (Barx1) was expressed at higher levels in cementoblasts, and immunohistochemistry indicated that BARX1 was expressed in apical cementoblasts and cementocytes, but not in osteoblasts or odontoblasts.
Conclusion
The OC‐GFP mouse provides a good model for selectively isolating cementoblasts, and allowed for identification of differentially expressed genes between cementoblasts and osteoblasts.</description><identifier>ISSN: 0022-3484</identifier><identifier>EISSN: 1600-0765</identifier><identifier>DOI: 10.1111/jre.12309</identifier><identifier>PMID: 26215316</identifier><language>eng</language><publisher>United States: Blackwell Publishing Ltd</publisher><subject>Animals ; Calcification, Physiologic ; Cell Differentiation - genetics ; cementoblast ; Dental Cementum - cytology ; Dental Cementum - drug effects ; Dental Cementum - physiology ; Dentistry ; Extracellular Matrix Proteins - pharmacology ; Fibroblasts - cytology ; gene expression ; Gene Expression Regulation ; Homeodomain Proteins - biosynthesis ; Homeodomain Proteins - genetics ; Intercellular Signaling Peptides and Proteins - pharmacology ; Membrane Proteins - pharmacology ; Mice ; Mice, Transgenic ; Odontoblasts - cytology ; osteoblast ; Osteoblasts - cytology ; Osteoblasts - drug effects ; Osteoblasts - physiology ; Osteocalcin ; Periodontal Ligament - cytology ; RNA, Messenger - genetics ; Tooth Root - cytology ; Transcription Factors - biosynthesis ; Transcription Factors - genetics ; Wnt signaling pathway ; Wnt Signaling Pathway - drug effects ; Wnt Signaling Pathway - genetics</subject><ispartof>Journal of periodontal research, 2016-06, Vol.51 (3), p.304-312</ispartof><rights>2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd</rights><rights>2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4869-78f3f9b9e9d25cf279c0af3968653b40f926e85f62fce5ae966e5ccfcbf330313</citedby><cites>FETCH-LOGICAL-c4869-78f3f9b9e9d25cf279c0af3968653b40f926e85f62fce5ae966e5ccfcbf330313</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fjre.12309$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fjre.12309$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>230,314,776,780,881,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26215316$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Matthews, B. G.</creatorcontrib><creatorcontrib>Roguljic, H.</creatorcontrib><creatorcontrib>Franceschetti, T.</creatorcontrib><creatorcontrib>Roeder, E.</creatorcontrib><creatorcontrib>Matic, I.</creatorcontrib><creatorcontrib>Vidovic, I.</creatorcontrib><creatorcontrib>Joshi, P.</creatorcontrib><creatorcontrib>Kum, K.-Y.</creatorcontrib><creatorcontrib>Kalajzic, I.</creatorcontrib><title>Gene-expression analysis of cementoblasts and osteoblasts</title><title>Journal of periodontal research</title><addtitle>J Periodont Res</addtitle><description>Background and Objective
Cementum and bone are similar mineralized tissues, but cementum accumulates much more slowly than bone, does not have vasculature or innervation and does not undergo remodeling. Despite these differences, there are no well‐established markers to distinguish cementoblasts from other mature mineralizing cells such as osteoblasts and odontoblasts. The purpose of this study was to assess differences in gene expression between cementoblasts and osteoblasts using gene profiling of cell populations isolated directly from osteocalcin–green fluorescent protein (OC‐GFP) transgenic mice.
Material and Methods
OC‐GFP reporter mice were used as they show labeling of cementoblasts, osteoblasts and odontoblasts, but not of periodontal ligament fibroblasts, within the periodontium. We sorted cells digested from the molar root surface to isolate OC‐GFP+ cementoblasts. Osteoblasts were isolated from calvarial digests. Microarray analysis was performed, and selected results were confirmed by real‐time PCR and immunostaining or in situ hybridization.
Results
Microarray analysis identified 95 genes that were expressed at least two‐fold higher in cementoblasts than in osteoblasts. Our analysis indicated that the Wnt signaling pathway was differentially regulated, as were genes related to skeletal development. Real‐time PCR confirmed that expression of the Wnt inhibitors Wnt inhibitory factor 1 (Wif1) and secreted frizzled‐related protein 1 (Sfrp1) was elevated in cementoblasts compared with osteoblasts, and Wif1 expression was localized to the apical root region. In addition, the transcription factor BARX homeobox 1 (Barx1) was expressed at higher levels in cementoblasts, and immunohistochemistry indicated that BARX1 was expressed in apical cementoblasts and cementocytes, but not in osteoblasts or odontoblasts.
Conclusion
The OC‐GFP mouse provides a good model for selectively isolating cementoblasts, and allowed for identification of differentially expressed genes between cementoblasts and osteoblasts.</description><subject>Animals</subject><subject>Calcification, Physiologic</subject><subject>Cell Differentiation - genetics</subject><subject>cementoblast</subject><subject>Dental Cementum - cytology</subject><subject>Dental Cementum - drug effects</subject><subject>Dental Cementum - physiology</subject><subject>Dentistry</subject><subject>Extracellular Matrix Proteins - pharmacology</subject><subject>Fibroblasts - cytology</subject><subject>gene expression</subject><subject>Gene Expression Regulation</subject><subject>Homeodomain Proteins - biosynthesis</subject><subject>Homeodomain Proteins - genetics</subject><subject>Intercellular Signaling Peptides and Proteins - pharmacology</subject><subject>Membrane Proteins - pharmacology</subject><subject>Mice</subject><subject>Mice, Transgenic</subject><subject>Odontoblasts - cytology</subject><subject>osteoblast</subject><subject>Osteoblasts - cytology</subject><subject>Osteoblasts - drug effects</subject><subject>Osteoblasts - physiology</subject><subject>Osteocalcin</subject><subject>Periodontal Ligament - cytology</subject><subject>RNA, Messenger - genetics</subject><subject>Tooth Root - cytology</subject><subject>Transcription Factors - biosynthesis</subject><subject>Transcription Factors - genetics</subject><subject>Wnt signaling pathway</subject><subject>Wnt Signaling Pathway - drug effects</subject><subject>Wnt Signaling Pathway - genetics</subject><issn>0022-3484</issn><issn>1600-0765</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUtLAzEQx4MoWh8Hv4D0qIfVPDbJ5iJo0fpCUXzdQjad6Op2U5NW7bc32lr0IOYShvnNjxn-CK0TvE3S23kKsE0ow2oOtYjAOMNS8HnUwpjSjOVFvoSWY3zCqRZSLaIlKijhjIgWUl1oIIP3QYAYK9-0TWPqcaxi27u2hT40Q1_WJg5j6vTaPg5hWq-iBWfqCGvTfwXdHB5cd46ys4vucWfvLLN5IVQmC8ecKhWoHuXWUaksNo4pUQjOyhw7RQUU3AnqLHADSgjg1jpbOsYwI2wF7U68g1HZh55NGwVT60Go-iaMtTeV_t1pqkf94F91LmmSqSTYnAqCfxlBHOp-FS3UtWnAj6ImBea5Ulzh_1FZ5JJjgUVCtyaoDT7GAG62EcH6MxWdUtFfqSR24-cJM_I7hgTsTIC3qobx3yZ9cnXwrcwmE1VK5H02YcKzFpJJru_Ou_rk-vK2c3q_rzn7AOc2ptU</recordid><startdate>201606</startdate><enddate>201606</enddate><creator>Matthews, B. G.</creator><creator>Roguljic, H.</creator><creator>Franceschetti, T.</creator><creator>Roeder, E.</creator><creator>Matic, I.</creator><creator>Vidovic, I.</creator><creator>Joshi, P.</creator><creator>Kum, K.-Y.</creator><creator>Kalajzic, I.</creator><general>Blackwell Publishing Ltd</general><scope>BSCLL</scope><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>7X8</scope><scope>7QP</scope><scope>5PM</scope></search><sort><creationdate>201606</creationdate><title>Gene-expression analysis of cementoblasts and osteoblasts</title><author>Matthews, B. G. ; Roguljic, H. ; Franceschetti, T. ; Roeder, E. ; Matic, I. ; Vidovic, I. ; Joshi, P. ; Kum, K.-Y. ; Kalajzic, I.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4869-78f3f9b9e9d25cf279c0af3968653b40f926e85f62fce5ae966e5ccfcbf330313</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Animals</topic><topic>Calcification, Physiologic</topic><topic>Cell Differentiation - genetics</topic><topic>cementoblast</topic><topic>Dental Cementum - cytology</topic><topic>Dental Cementum - drug effects</topic><topic>Dental Cementum - physiology</topic><topic>Dentistry</topic><topic>Extracellular Matrix Proteins - pharmacology</topic><topic>Fibroblasts - cytology</topic><topic>gene expression</topic><topic>Gene Expression Regulation</topic><topic>Homeodomain Proteins - biosynthesis</topic><topic>Homeodomain Proteins - genetics</topic><topic>Intercellular Signaling Peptides and Proteins - pharmacology</topic><topic>Membrane Proteins - pharmacology</topic><topic>Mice</topic><topic>Mice, Transgenic</topic><topic>Odontoblasts - cytology</topic><topic>osteoblast</topic><topic>Osteoblasts - cytology</topic><topic>Osteoblasts - drug effects</topic><topic>Osteoblasts - physiology</topic><topic>Osteocalcin</topic><topic>Periodontal Ligament - cytology</topic><topic>RNA, Messenger - genetics</topic><topic>Tooth Root - cytology</topic><topic>Transcription Factors - biosynthesis</topic><topic>Transcription Factors - genetics</topic><topic>Wnt signaling pathway</topic><topic>Wnt Signaling Pathway - drug effects</topic><topic>Wnt Signaling Pathway - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Matthews, B. G.</creatorcontrib><creatorcontrib>Roguljic, H.</creatorcontrib><creatorcontrib>Franceschetti, T.</creatorcontrib><creatorcontrib>Roeder, E.</creatorcontrib><creatorcontrib>Matic, I.</creatorcontrib><creatorcontrib>Vidovic, I.</creatorcontrib><creatorcontrib>Joshi, P.</creatorcontrib><creatorcontrib>Kum, K.-Y.</creatorcontrib><creatorcontrib>Kalajzic, I.</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of periodontal research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Matthews, B. G.</au><au>Roguljic, H.</au><au>Franceschetti, T.</au><au>Roeder, E.</au><au>Matic, I.</au><au>Vidovic, I.</au><au>Joshi, P.</au><au>Kum, K.-Y.</au><au>Kalajzic, I.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Gene-expression analysis of cementoblasts and osteoblasts</atitle><jtitle>Journal of periodontal research</jtitle><addtitle>J Periodont Res</addtitle><date>2016-06</date><risdate>2016</risdate><volume>51</volume><issue>3</issue><spage>304</spage><epage>312</epage><pages>304-312</pages><issn>0022-3484</issn><eissn>1600-0765</eissn><abstract>Background and Objective
Cementum and bone are similar mineralized tissues, but cementum accumulates much more slowly than bone, does not have vasculature or innervation and does not undergo remodeling. Despite these differences, there are no well‐established markers to distinguish cementoblasts from other mature mineralizing cells such as osteoblasts and odontoblasts. The purpose of this study was to assess differences in gene expression between cementoblasts and osteoblasts using gene profiling of cell populations isolated directly from osteocalcin–green fluorescent protein (OC‐GFP) transgenic mice.
Material and Methods
OC‐GFP reporter mice were used as they show labeling of cementoblasts, osteoblasts and odontoblasts, but not of periodontal ligament fibroblasts, within the periodontium. We sorted cells digested from the molar root surface to isolate OC‐GFP+ cementoblasts. Osteoblasts were isolated from calvarial digests. Microarray analysis was performed, and selected results were confirmed by real‐time PCR and immunostaining or in situ hybridization.
Results
Microarray analysis identified 95 genes that were expressed at least two‐fold higher in cementoblasts than in osteoblasts. Our analysis indicated that the Wnt signaling pathway was differentially regulated, as were genes related to skeletal development. Real‐time PCR confirmed that expression of the Wnt inhibitors Wnt inhibitory factor 1 (Wif1) and secreted frizzled‐related protein 1 (Sfrp1) was elevated in cementoblasts compared with osteoblasts, and Wif1 expression was localized to the apical root region. In addition, the transcription factor BARX homeobox 1 (Barx1) was expressed at higher levels in cementoblasts, and immunohistochemistry indicated that BARX1 was expressed in apical cementoblasts and cementocytes, but not in osteoblasts or odontoblasts.
Conclusion
The OC‐GFP mouse provides a good model for selectively isolating cementoblasts, and allowed for identification of differentially expressed genes between cementoblasts and osteoblasts.</abstract><cop>United States</cop><pub>Blackwell Publishing Ltd</pub><pmid>26215316</pmid><doi>10.1111/jre.12309</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0022-3484 |
| ispartof | Journal of periodontal research, 2016-06, Vol.51 (3), p.304-312 |
| issn | 0022-3484 1600-0765 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4729669 |
| source | MEDLINE; Wiley Online Library Journals Frontfile Complete |
| subjects | Animals Calcification, Physiologic Cell Differentiation - genetics cementoblast Dental Cementum - cytology Dental Cementum - drug effects Dental Cementum - physiology Dentistry Extracellular Matrix Proteins - pharmacology Fibroblasts - cytology gene expression Gene Expression Regulation Homeodomain Proteins - biosynthesis Homeodomain Proteins - genetics Intercellular Signaling Peptides and Proteins - pharmacology Membrane Proteins - pharmacology Mice Mice, Transgenic Odontoblasts - cytology osteoblast Osteoblasts - cytology Osteoblasts - drug effects Osteoblasts - physiology Osteocalcin Periodontal Ligament - cytology RNA, Messenger - genetics Tooth Root - cytology Transcription Factors - biosynthesis Transcription Factors - genetics Wnt signaling pathway Wnt Signaling Pathway - drug effects Wnt Signaling Pathway - genetics |
| title | Gene-expression analysis of cementoblasts and osteoblasts |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-21T15%3A08%3A43IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Gene-expression%20analysis%20of%20cementoblasts%20and%20osteoblasts&rft.jtitle=Journal%20of%20periodontal%20research&rft.au=Matthews,%20B.%20G.&rft.date=2016-06&rft.volume=51&rft.issue=3&rft.spage=304&rft.epage=312&rft.pages=304-312&rft.issn=0022-3484&rft.eissn=1600-0765&rft_id=info:doi/10.1111/jre.12309&rft_dat=%3Cproquest_pubme%3E1784750606%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1784750606&rft_id=info:pmid/26215316&rfr_iscdi=true |