Effect of Enamel Matrix Derivative on the Differentiation of C2C12 Cells
Background: Although enamel matrix derivative (EMD) can initiate de novo cementum and bone formation by stimulating and inducing differentiation of mesenchymal cells in the periodontal ligament, the molecular mechanism of this phenomenon is not fully understood. The purpose of this study was to dete...
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| Veröffentlicht in: | Journal of periodontology (1970) 2002-05, Vol.73 (5), p.543-550 |
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| creator | Ohyama, Mariko Suzuki, Naoto Yamaguchi, Yoko Maeno, Masao Otsuka, Kichibee Ito, Koichi |
| description | Background: Although enamel matrix derivative (EMD) can initiate de novo cementum and bone formation by stimulating and inducing differentiation of mesenchymal cells in the periodontal ligament, the molecular mechanism of this phenomenon is not fully understood. The purpose of this study was to determine the effect of EMD on the differentiation of pluripotential mesenchymal cells.
Methods: A typical pluripotential mesenchymal cell line, C2C12, was used to clarify the effect of EMD on cell differentiation. The cells were cultured in 5% serum‐containing medium to induce cell differentiation, either with or without the addition of EMD. Differentiation to myoblasts was analyzed by immunostaining of desmin and type II myosin heavy chains. Osteoblast differentiation was evaluated by measuring alkaline phosphatase (ALPase) activity. Furthermore, to verify the cell lineage after culture with EMD, mRNA expression of cellular phenotypespecific markers characterizing osteoblasts (ALPase and osteocalcin), chondroblasts (type X collagen), myoblasts (desmin and MyoD), and adipocytes (lipoprotein lipase) was studied using semiquantitative reverse transcription‐polymerase chain reaction.
Results: C2C12 cells cultured in differentiation medium without EMD altered their phenotype to myoblasts, exhibiting positive reactions to desmin and myosin heavy chains by immunological analysis. However, the cells cultured in the presence of EMD were strongly inhibited from developing into myoblasts, and showed high ALPase activity that was approximately 2 to 4 times greater than that of the vehicle. The mRNA expression of ALPase, osteocalcin, and type X collagen was increased markedly by the EMD‐stimulated medium, whereas the expression of desmin, MyoD, and lipoprotein lipase was drastically decreased.
Conclusions: Our study provides clear evidence that EMD converts the differentiation pathway of C2C12 cells into the osteoblast and/or chondroblast lineage. J Periodontol 2002;73:543‐550. |
| doi_str_mv | 10.1902/jop.2002.73.5.543 |
| format | Article |
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Methods: A typical pluripotential mesenchymal cell line, C2C12, was used to clarify the effect of EMD on cell differentiation. The cells were cultured in 5% serum‐containing medium to induce cell differentiation, either with or without the addition of EMD. Differentiation to myoblasts was analyzed by immunostaining of desmin and type II myosin heavy chains. Osteoblast differentiation was evaluated by measuring alkaline phosphatase (ALPase) activity. Furthermore, to verify the cell lineage after culture with EMD, mRNA expression of cellular phenotypespecific markers characterizing osteoblasts (ALPase and osteocalcin), chondroblasts (type X collagen), myoblasts (desmin and MyoD), and adipocytes (lipoprotein lipase) was studied using semiquantitative reverse transcription‐polymerase chain reaction.
Results: C2C12 cells cultured in differentiation medium without EMD altered their phenotype to myoblasts, exhibiting positive reactions to desmin and myosin heavy chains by immunological analysis. However, the cells cultured in the presence of EMD were strongly inhibited from developing into myoblasts, and showed high ALPase activity that was approximately 2 to 4 times greater than that of the vehicle. The mRNA expression of ALPase, osteocalcin, and type X collagen was increased markedly by the EMD‐stimulated medium, whereas the expression of desmin, MyoD, and lipoprotein lipase was drastically decreased.
Conclusions: Our study provides clear evidence that EMD converts the differentiation pathway of C2C12 cells into the osteoblast and/or chondroblast lineage. J Periodontol 2002;73:543‐550.</description><identifier>ISSN: 0022-3492</identifier><identifier>EISSN: 1943-3670</identifier><identifier>DOI: 10.1902/jop.2002.73.5.543</identifier><identifier>PMID: 12027258</identifier><language>eng</language><publisher>737 N. Michigan Avenue, Suite 800, Chicago, IL 60611‐2690, USA: American Academy of Periodontology</publisher><subject>Adipocytes - cytology ; Adipocytes - drug effects ; Alkaline Phosphatase - analysis ; Animals ; Biomarkers - analysis ; cell differentiation ; Cell Differentiation - drug effects ; Cell Division - drug effects ; Cell Line ; Cell Lineage ; Chondrocytes - cytology ; Chondrocytes - drug effects ; Collagen Type X - analysis ; Culture Media ; Dental Enamel Proteins - pharmacology ; Dentistry ; Desmin - analysis ; Enamel matrix derivative ; Lipoprotein Lipase - analysis ; Mesoderm - cytology ; Mesoderm - drug effects ; Mice ; Muscle Fibers, Skeletal - cytology ; Muscle Fibers, Skeletal - drug effects ; myoblasts ; MyoD Protein - analysis ; Myosin Heavy Chains - analysis ; osteoblasts ; Osteoblasts - cytology ; Osteoblasts - drug effects ; Osteocalcin - analysis ; Phenotype ; Statistics as Topic ; Stem Cells - drug effects</subject><ispartof>Journal of periodontology (1970), 2002-05, Vol.73 (5), p.543-550</ispartof><rights>2002 American Academy of Periodontology</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4113-a37f4ce263fb249197ad65caacdf03db37590e1d65a308e32137388a8b3f8b713</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1902%2Fjop.2002.73.5.543$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1902%2Fjop.2002.73.5.543$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12027258$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ohyama, Mariko</creatorcontrib><creatorcontrib>Suzuki, Naoto</creatorcontrib><creatorcontrib>Yamaguchi, Yoko</creatorcontrib><creatorcontrib>Maeno, Masao</creatorcontrib><creatorcontrib>Otsuka, Kichibee</creatorcontrib><creatorcontrib>Ito, Koichi</creatorcontrib><title>Effect of Enamel Matrix Derivative on the Differentiation of C2C12 Cells</title><title>Journal of periodontology (1970)</title><addtitle>J Periodontol</addtitle><description>Background: Although enamel matrix derivative (EMD) can initiate de novo cementum and bone formation by stimulating and inducing differentiation of mesenchymal cells in the periodontal ligament, the molecular mechanism of this phenomenon is not fully understood. The purpose of this study was to determine the effect of EMD on the differentiation of pluripotential mesenchymal cells.
Methods: A typical pluripotential mesenchymal cell line, C2C12, was used to clarify the effect of EMD on cell differentiation. The cells were cultured in 5% serum‐containing medium to induce cell differentiation, either with or without the addition of EMD. Differentiation to myoblasts was analyzed by immunostaining of desmin and type II myosin heavy chains. Osteoblast differentiation was evaluated by measuring alkaline phosphatase (ALPase) activity. Furthermore, to verify the cell lineage after culture with EMD, mRNA expression of cellular phenotypespecific markers characterizing osteoblasts (ALPase and osteocalcin), chondroblasts (type X collagen), myoblasts (desmin and MyoD), and adipocytes (lipoprotein lipase) was studied using semiquantitative reverse transcription‐polymerase chain reaction.
Results: C2C12 cells cultured in differentiation medium without EMD altered their phenotype to myoblasts, exhibiting positive reactions to desmin and myosin heavy chains by immunological analysis. However, the cells cultured in the presence of EMD were strongly inhibited from developing into myoblasts, and showed high ALPase activity that was approximately 2 to 4 times greater than that of the vehicle. The mRNA expression of ALPase, osteocalcin, and type X collagen was increased markedly by the EMD‐stimulated medium, whereas the expression of desmin, MyoD, and lipoprotein lipase was drastically decreased.
Conclusions: Our study provides clear evidence that EMD converts the differentiation pathway of C2C12 cells into the osteoblast and/or chondroblast lineage. J Periodontol 2002;73:543‐550.</description><subject>Adipocytes - cytology</subject><subject>Adipocytes - drug effects</subject><subject>Alkaline Phosphatase - analysis</subject><subject>Animals</subject><subject>Biomarkers - analysis</subject><subject>cell differentiation</subject><subject>Cell Differentiation - drug effects</subject><subject>Cell Division - drug effects</subject><subject>Cell Line</subject><subject>Cell Lineage</subject><subject>Chondrocytes - cytology</subject><subject>Chondrocytes - drug effects</subject><subject>Collagen Type X - analysis</subject><subject>Culture Media</subject><subject>Dental Enamel Proteins - pharmacology</subject><subject>Dentistry</subject><subject>Desmin - analysis</subject><subject>Enamel matrix derivative</subject><subject>Lipoprotein Lipase - analysis</subject><subject>Mesoderm - cytology</subject><subject>Mesoderm - drug effects</subject><subject>Mice</subject><subject>Muscle Fibers, Skeletal - cytology</subject><subject>Muscle Fibers, Skeletal - drug effects</subject><subject>myoblasts</subject><subject>MyoD Protein - analysis</subject><subject>Myosin Heavy Chains - analysis</subject><subject>osteoblasts</subject><subject>Osteoblasts - cytology</subject><subject>Osteoblasts - drug effects</subject><subject>Osteocalcin - analysis</subject><subject>Phenotype</subject><subject>Statistics as Topic</subject><subject>Stem Cells - drug effects</subject><issn>0022-3492</issn><issn>1943-3670</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE9PwyAchonRuDn9AF4MJ2-twI-W9mhqdZoZjdEzoS1Elv6Z0E337WXZEo-eCC_P-4Y8CF1SEtOcsJvlsIoZISwWECdxwuEITWnOIYJUkGM0DU8sAp6zCTrzfhmulAM5RRPKCBMsyaZoXhqj6xEPBpe96nSLn9Xo7A--085u1Gg3Gg89Hj81vrMBdbofbYhDFioFKyjDhW5bf45OjGq9vjicM_RxX74X82jx8vBY3C6imlMKkQJheK1ZCqZiPKe5UE2a1ErVjSHQVCCSnGgaMgUk08AoCMgylVVgskpQmKHr_e7KDV9r7UfZWV-HH6heD2svBRWUM54GkO7B2g3eO23kytlOua2kRO70yaBP7vRJATKRQV_oXB3G11Wnm7_GwVcAxB74tq3e_r8on17LN7Kb_gUnNXrA</recordid><startdate>200205</startdate><enddate>200205</enddate><creator>Ohyama, Mariko</creator><creator>Suzuki, Naoto</creator><creator>Yamaguchi, Yoko</creator><creator>Maeno, Masao</creator><creator>Otsuka, Kichibee</creator><creator>Ito, Koichi</creator><general>American Academy of Periodontology</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>7X8</scope></search><sort><creationdate>200205</creationdate><title>Effect of Enamel Matrix Derivative on the Differentiation of C2C12 Cells</title><author>Ohyama, Mariko ; Suzuki, Naoto ; Yamaguchi, Yoko ; Maeno, Masao ; Otsuka, Kichibee ; Ito, Koichi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4113-a37f4ce263fb249197ad65caacdf03db37590e1d65a308e32137388a8b3f8b713</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Adipocytes - cytology</topic><topic>Adipocytes - drug effects</topic><topic>Alkaline Phosphatase - analysis</topic><topic>Animals</topic><topic>Biomarkers - analysis</topic><topic>cell differentiation</topic><topic>Cell Differentiation - drug effects</topic><topic>Cell Division - drug effects</topic><topic>Cell Line</topic><topic>Cell Lineage</topic><topic>Chondrocytes - cytology</topic><topic>Chondrocytes - drug effects</topic><topic>Collagen Type X - analysis</topic><topic>Culture Media</topic><topic>Dental Enamel Proteins - pharmacology</topic><topic>Dentistry</topic><topic>Desmin - analysis</topic><topic>Enamel matrix derivative</topic><topic>Lipoprotein Lipase - analysis</topic><topic>Mesoderm - cytology</topic><topic>Mesoderm - drug effects</topic><topic>Mice</topic><topic>Muscle Fibers, Skeletal - cytology</topic><topic>Muscle Fibers, Skeletal - drug effects</topic><topic>myoblasts</topic><topic>MyoD Protein - analysis</topic><topic>Myosin Heavy Chains - analysis</topic><topic>osteoblasts</topic><topic>Osteoblasts - cytology</topic><topic>Osteoblasts - drug effects</topic><topic>Osteocalcin - analysis</topic><topic>Phenotype</topic><topic>Statistics as Topic</topic><topic>Stem Cells - drug effects</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ohyama, Mariko</creatorcontrib><creatorcontrib>Suzuki, Naoto</creatorcontrib><creatorcontrib>Yamaguchi, Yoko</creatorcontrib><creatorcontrib>Maeno, Masao</creatorcontrib><creatorcontrib>Otsuka, Kichibee</creatorcontrib><creatorcontrib>Ito, Koichi</creatorcontrib><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><jtitle>Journal of periodontology (1970)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ohyama, Mariko</au><au>Suzuki, Naoto</au><au>Yamaguchi, Yoko</au><au>Maeno, Masao</au><au>Otsuka, Kichibee</au><au>Ito, Koichi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Effect of Enamel Matrix Derivative on the Differentiation of C2C12 Cells</atitle><jtitle>Journal of periodontology (1970)</jtitle><addtitle>J Periodontol</addtitle><date>2002-05</date><risdate>2002</risdate><volume>73</volume><issue>5</issue><spage>543</spage><epage>550</epage><pages>543-550</pages><issn>0022-3492</issn><eissn>1943-3670</eissn><abstract>Background: Although enamel matrix derivative (EMD) can initiate de novo cementum and bone formation by stimulating and inducing differentiation of mesenchymal cells in the periodontal ligament, the molecular mechanism of this phenomenon is not fully understood. The purpose of this study was to determine the effect of EMD on the differentiation of pluripotential mesenchymal cells.
Methods: A typical pluripotential mesenchymal cell line, C2C12, was used to clarify the effect of EMD on cell differentiation. The cells were cultured in 5% serum‐containing medium to induce cell differentiation, either with or without the addition of EMD. Differentiation to myoblasts was analyzed by immunostaining of desmin and type II myosin heavy chains. Osteoblast differentiation was evaluated by measuring alkaline phosphatase (ALPase) activity. Furthermore, to verify the cell lineage after culture with EMD, mRNA expression of cellular phenotypespecific markers characterizing osteoblasts (ALPase and osteocalcin), chondroblasts (type X collagen), myoblasts (desmin and MyoD), and adipocytes (lipoprotein lipase) was studied using semiquantitative reverse transcription‐polymerase chain reaction.
Results: C2C12 cells cultured in differentiation medium without EMD altered their phenotype to myoblasts, exhibiting positive reactions to desmin and myosin heavy chains by immunological analysis. However, the cells cultured in the presence of EMD were strongly inhibited from developing into myoblasts, and showed high ALPase activity that was approximately 2 to 4 times greater than that of the vehicle. The mRNA expression of ALPase, osteocalcin, and type X collagen was increased markedly by the EMD‐stimulated medium, whereas the expression of desmin, MyoD, and lipoprotein lipase was drastically decreased.
Conclusions: Our study provides clear evidence that EMD converts the differentiation pathway of C2C12 cells into the osteoblast and/or chondroblast lineage. J Periodontol 2002;73:543‐550.</abstract><cop>737 N. Michigan Avenue, Suite 800, Chicago, IL 60611‐2690, USA</cop><pub>American Academy of Periodontology</pub><pmid>12027258</pmid><doi>10.1902/jop.2002.73.5.543</doi><tpages>8</tpages></addata></record> |
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| subjects | Adipocytes - cytology Adipocytes - drug effects Alkaline Phosphatase - analysis Animals Biomarkers - analysis cell differentiation Cell Differentiation - drug effects Cell Division - drug effects Cell Line Cell Lineage Chondrocytes - cytology Chondrocytes - drug effects Collagen Type X - analysis Culture Media Dental Enamel Proteins - pharmacology Dentistry Desmin - analysis Enamel matrix derivative Lipoprotein Lipase - analysis Mesoderm - cytology Mesoderm - drug effects Mice Muscle Fibers, Skeletal - cytology Muscle Fibers, Skeletal - drug effects myoblasts MyoD Protein - analysis Myosin Heavy Chains - analysis osteoblasts Osteoblasts - cytology Osteoblasts - drug effects Osteocalcin - analysis Phenotype Statistics as Topic Stem Cells - drug effects |
| title | Effect of Enamel Matrix Derivative on the Differentiation of C2C12 Cells |
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