Bone marrow mesenchymal stem cells combine with Treated dentin matrix to build biological root
Identifieur interne : 000304 ( Pmc/Checkpoint ); précédent : 000303; suivant : 000305Bone marrow mesenchymal stem cells combine with Treated dentin matrix to build biological root
Auteurs : Shiwei Luo [République populaire de Chine] ; Fei Pei [République populaire de Chine] ; Wen Zhang [République populaire de Chine] ; Weihua Guo [République populaire de Chine] ; Rui Li [République populaire de Chine] ; Wei He [République populaire de Chine] ; Weidong Tian [République populaire de Chine]Source :
- Scientific Reports [ 2045-2322 ] ; 2017.
Abstract
Treated dentin matrix (TDM) as a kind of scaffolding material has been proved odontogenic induction ability on dental-derived stem cells. Given the limited resources of dental stem cells, it is necessary to seek new seed cell which easily obtained. Jaw bone marrow mesenchymal stem cell (JBMMSC) as non-dental-derived stem cell relates to the development of teeth and jaws which suggest us JBMMSCs could act as a new seed cell for tooth tissue engineering. To assess the odontogenic and osteogenic potential of JBMMSCs, cells were induced by TDM extraction
Url:
DOI: 10.1038/srep44635
PubMed: 28401887
PubMed Central: 5388852
Affiliations:
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Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a3"><institution>State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Zhang, Wen" sort="Zhang, Wen" uniqKey="Zhang W" first="Wen" last="Zhang">Wen Zhang</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University</institution>, Zhengzhou,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Guo, Weihua" sort="Guo, Weihua" uniqKey="Guo W" first="Weihua" last="Guo">Weihua Guo</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>National Engineering Laboratory for Oral Regenerative Medicine, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a3"><institution>State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a4"><institution>Department of Pedodontics, West China School of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Li, Rui" sort="Li, Rui" uniqKey="Li R" first="Rui" last="Li">Rui Li</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University</institution>, Zhengzhou,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="He, Wei" sort="He, Wei" uniqKey="He W" first="Wei" last="He">Wei He</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University</institution>, Zhengzhou,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Tian, Weidong" sort="Tian, Weidong" uniqKey="Tian W" first="Weidong" last="Tian">Weidong Tian</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>National Engineering Laboratory for Oral Regenerative Medicine, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a3"><institution>State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a5"><institution>Department of Oral and Maxillofacial Surgery, West China School of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">28401887</idno><idno type="pmc">5388852</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5388852</idno><idno type="RBID">PMC:5388852</idno><idno type="doi">10.1038/srep44635</idno><date when="2017">2017</date><idno type="wicri:Area/Pmc/Corpus">000847</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000847</idno><idno type="wicri:Area/Pmc/Curation">000847</idno><idno type="wicri:explorRef" 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Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a3"><institution>State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Pei, Fei" sort="Pei, Fei" uniqKey="Pei F" first="Fei" last="Pei">Fei Pei</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University</institution>, Zhengzhou,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a2"><institution>National Engineering Laboratory for Oral Regenerative Medicine, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a3"><institution>State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Zhang, Wen" sort="Zhang, Wen" uniqKey="Zhang W" first="Wen" last="Zhang">Wen Zhang</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University</institution>, Zhengzhou,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Guo, Weihua" sort="Guo, Weihua" uniqKey="Guo W" first="Weihua" last="Guo">Weihua Guo</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>National Engineering Laboratory for Oral Regenerative Medicine, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a3"><institution>State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a4"><institution>Department of Pedodontics, West China School of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Li, Rui" sort="Li, Rui" uniqKey="Li R" first="Rui" last="Li">Rui Li</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University</institution>, Zhengzhou,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="He, Wei" sort="He, Wei" uniqKey="He W" first="Wei" last="He">Wei He</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University</institution>, Zhengzhou,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Tian, Weidong" sort="Tian, Weidong" uniqKey="Tian W" first="Weidong" last="Tian">Weidong Tian</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>National Engineering Laboratory for Oral Regenerative Medicine, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a3"><institution>State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a5"><institution>Department of Oral and Maxillofacial Surgery, West China School of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></analytic><series><title level="j">Scientific Reports</title><idno type="eISSN">2045-2322</idno><imprint><date when="2017">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Treated dentin matrix (TDM) as a kind of scaffolding material has been proved odontogenic induction ability on dental-derived stem cells. Given the limited resources of dental stem cells, it is necessary to seek new seed cell which easily obtained. Jaw bone marrow mesenchymal stem cell (JBMMSC) as non-dental-derived stem cell relates to the development of teeth and jaws which suggest us JBMMSCs could act as a new seed cell for tooth tissue engineering. To assess the odontogenic and osteogenic potential of JBMMSCs, cells were induced by TDM extraction <italic>in vitro</italic> and combined with TDM <italic>in vivo</italic>. Results were analyzed by PCR, Western Blotting and histology. PCR and Western Blotting showed odontogenic and osteogenic makers were significantly enhanced in varying degrees after induced by TDM extraction <italic>in vitro. In vivo</italic>, JBMMSCs expressed both odontogenic and osteogenic-related protein, and the latter showed stronger positive expression. Furthermore, histological examination of the harvested grafts was observed the formation of bone-like tissue. Therefore, osteogenic differentiation ability of JBMMSCs were enhanced significantly after being inducted by TDM which illustrates that non-odontogenic derived stem cells are still promising seed cells in tooth root tissue engineering.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Mao, J J" uniqKey="Mao J">J. J. Mao</name></author><author><name sortKey="Prockop, D J" uniqKey="Prockop D">D. J. Prockop</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Fugazzotto, P A" uniqKey="Fugazzotto P">P. A. Fugazzotto</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Li, R" uniqKey="Li R">R. Li</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Guo, W" uniqKey="Guo W">W. Guo</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yen, A H" uniqKey="Yen A">A. H. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Sci Rep</journal-id><journal-id journal-id-type="iso-abbrev">Sci Rep</journal-id><journal-title-group><journal-title>Scientific Reports</journal-title></journal-title-group><issn pub-type="epub">2045-2322</issn><publisher><publisher-name>Nature Publishing Group</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">28401887</article-id><article-id pub-id-type="pmc">5388852</article-id><article-id pub-id-type="pii">srep44635</article-id><article-id pub-id-type="doi">10.1038/srep44635</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Bone marrow mesenchymal stem cells combine with Treated dentin matrix to build biological root</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Luo</surname><given-names>Shiwei</given-names></name><xref ref-type="aff" rid="a1">1</xref><xref ref-type="aff" rid="a2">2</xref><xref ref-type="aff" rid="a3">3</xref><xref ref-type="author-notes" rid="n1">*</xref></contrib><contrib contrib-type="author"><name><surname>Pei</surname><given-names>Fei</given-names></name><xref ref-type="aff" rid="a1">1</xref><xref ref-type="aff" rid="a2">2</xref><xref ref-type="aff" rid="a3">3</xref><xref ref-type="author-notes" rid="n1">*</xref></contrib><contrib contrib-type="author"><name><surname>Zhang</surname><given-names>Wen</given-names></name><xref ref-type="aff" rid="a1">1</xref></contrib><contrib contrib-type="author"><name><surname>Guo</surname><given-names>Weihua</given-names></name><xref ref-type="aff" rid="a2">2</xref><xref ref-type="aff" rid="a3">3</xref><xref ref-type="aff" rid="a4">4</xref></contrib><contrib contrib-type="author"><name><surname>Li</surname><given-names>Rui</given-names></name><xref ref-type="corresp" rid="c1">a</xref><xref ref-type="aff" rid="a1">1</xref></contrib><contrib contrib-type="author"><name><surname>He</surname><given-names>Wei</given-names></name><xref ref-type="corresp" rid="c2">b</xref><xref ref-type="aff" rid="a1">1</xref></contrib><contrib contrib-type="author"><name><surname>Tian</surname><given-names>Weidong</given-names></name><xref ref-type="corresp" rid="c3">c</xref><xref ref-type="aff" rid="a2">2</xref><xref ref-type="aff" rid="a3">3</xref><xref ref-type="aff" rid="a5">5</xref></contrib><aff id="a1"><label>1</label><institution>Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University</institution>, Zhengzhou,<country>China</country></aff><aff id="a2"><label>2</label><institution>National Engineering Laboratory for Oral Regenerative Medicine, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></aff><aff id="a3"><label>3</label><institution>State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></aff><aff id="a4"><label>4</label><institution>Department of Pedodontics, West China School of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></aff><aff id="a5"><label>5</label><institution>Department of Oral and Maxillofacial Surgery, West China School of Stomatology, Sichuan University</institution>, Chengdu,<country>China</country></aff></contrib-group><author-notes><corresp id="c1"><label>a</label><email>zzulirui@126.com</email></corresp><corresp id="c2"><label>b</label><email>hewei1968@tom.com</email></corresp><corresp id="c3"><label>c</label><email>drtwd@sina.com</email></corresp><fn id="n1"><label>*</label><p>These authors contributed equally to this work.</p></fn></author-notes><pub-date pub-type="epub"><day>12</day><month>04</month><year>2017</year></pub-date><pub-date pub-type="collection"><year>2017</year></pub-date><volume>7</volume><elocation-id>44635</elocation-id><history><date date-type="received"><day>15</day><month>09</month><year>2016</year></date><date date-type="accepted"><day>13</day><month>02</month><year>2017</year></date></history><permissions><copyright-statement>Copyright © 2017, The Author(s)</copyright-statement><copyright-year>2017</copyright-year><copyright-holder>The Author(s)</copyright-holder><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by/4.0/"><pmc-comment>author-paid</pmc-comment>
<license-p>This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link></license-p></license></permissions><abstract><p>Treated dentin matrix (TDM) as a kind of scaffolding material has been proved odontogenic induction ability on dental-derived stem cells. Given the limited resources of dental stem cells, it is necessary to seek new seed cell which easily obtained. Jaw bone marrow mesenchymal stem cell (JBMMSC) as non-dental-derived stem cell relates to the development of teeth and jaws which suggest us JBMMSCs could act as a new seed cell for tooth tissue engineering. To assess the odontogenic and osteogenic potential of JBMMSCs, cells were induced by TDM extraction <italic>in vitro</italic> and combined with TDM <italic>in vivo</italic>. Results were analyzed by PCR, Western Blotting and histology. PCR and Western Blotting showed odontogenic and osteogenic makers were significantly enhanced in varying degrees after induced by TDM extraction <italic>in vitro. In vivo</italic>, JBMMSCs expressed both odontogenic and osteogenic-related protein, and the latter showed stronger positive expression. Furthermore, histological examination of the harvested grafts was observed the formation of bone-like tissue. Therefore, osteogenic differentiation ability of JBMMSCs were enhanced significantly after being inducted by TDM which illustrates that non-odontogenic derived stem cells are still promising seed cells in tooth root tissue engineering.</p></abstract></article-meta></front><floats-group><fig id="f1"><label>Figure 1</label><caption><title>Identification and characteristics of JBMMSCs and TDM.</title><p>Second-passage cells were typical spindle or triangle shape morphology (<bold>A</bold>) and positive for Vimentin but negative for CK14 in immunofluorescence staining (<bold>D</bold>). TDM (<bold>B</bold>) as scaffold was fabricated from human healthy teeth. SEM (<bold>C</bold>) illustrates that dentinal tubules exposed sufficiently and loosened fiber bundles of intertubular and peritubular dentin. (<bold>E</bold>) JBMMSCs were planted on the surface of TDM. Scale bars = 100 μm, 10 μm for (<bold>A</bold>,<bold>D)</bold> respectively.</p></caption><graphic xlink:href="srep44635-f1"></graphic></fig><fig id="f2"><label>Figure 2</label><caption><title>Characteristics of JBMMSCs.</title><p>Panel shows lipid clusters (<bold>A</bold>) mineralized nodules (<bold>B</bold>) were observed respectively when JBMMSCs were cultured in osteogenic, adipogenic inductive medium for 2 and 4 week. JBMMSCs were transformed into neuron-like cells (<bold>C</bold>) and vascular endothelial-like cells (<bold>E</bold>) in neurogenic and vascular inductive medium. Moreover, these neuron-like cells stained positively for nestin (<bold>D</bold>) and vascular endothelial-like cells were positive for CD31 (<bold>F</bold>) in immunofluorescence. (<bold>G</bold>) Flow cytometry indicated that JBMMSs were positive for mesenchymal stem cell surface marker CD29, CD73, CD90, CD105, CD146 while negative for platelet endothelial cell.adhesion molecule-1 CD31 and hematopietic stem cell marker CD 34. Scale bars = 50 μm for (<bold>A</bold>–<bold>D</bold>,<bold>F</bold>), Scale bars = 200 μm for (<bold>E</bold>).</p></caption><graphic xlink:href="srep44635-f2"></graphic></fig><fig id="f3"><label>Figure 3</label><caption><p>Real-time qPCR (<bold>A</bold>) and Western Blotting (<bold>B</bold>) were performed to assess the effect of the liquid extraction of TDM on the JBMMSCs at 3 d, 7 d, 14 d respectively. Overall, (<bold>A</bold>) TDM up-regulated the expression of odontogensis markers DSPP and DMP-1, osteogenesis marker Periostin and OCN, early stage osteogenic differentiation marker ALP and dentin noncollagen protein Decorin than control group but kept the expression level of Fibronectin. Meanwhile, the expression of COL-III was down-regulated after inducement (<bold>B</bold>) Western Blotting showed JBMMSCs expressed the odontogenesis related protein DMP1 and osteogenesis related protein OPN, OCN, CAP, Periostin, and RUNX2 stronger in varying degrees than control group at later time points. Interestingly, DSPP and Decorin maintained a stable expression as well as the control group. (*p < 0.05, **p < 0.01).</p></caption><graphic xlink:href="srep44635-f3"></graphic></fig><fig id="f4"><label>Figure 4</label><caption><p>H&E staining showed planted cells had ideal adherence and proliferation on TDM (<bold>A</bold>). JBMMSCs peformed connective tissue-like construction with new vessel generation (<bold>B</bold>). However expected regenerated dentin such as dentinal tubules, predentin, was not observed which is same in control group (single TDM) (<bold>D</bold>). Masson stain (<bold>C</bold>) showed the fibrous tissue performed. Scale bars = 200 μm for (<bold>A</bold>,<bold>D</bold>). Scale bars = 50 μm for (<bold>B</bold>,<bold>C</bold>).</p></caption><graphic xlink:href="srep44635-f4"></graphic></fig><fig id="f5"><label>Figure 5</label><caption><title>Immunohistochemistry was used to evaluate the expression of DSPP, DMP-1, OCN, OPN, Decorin, TGF-β, COL-I, COL-III, Periostin, Fibronectin and ALP to explored the differentiation status of JBMMSCs.</title><p>The result showed that JBMMSCs were strongly positive for protein osteogenic marker OPN (<bold>D</bold>) and ALP (<bold>F</bold>) not only in the cells but also in demineralized layer and dentin. Positive expression of OCN (<bold>E</bold>), Decorin (<bold>C</bold>) and Periostin (<bold>H</bold>) were also observed in cells and demineralized layer. Odontogenic markers DSPP (<bold>A</bold>), DMP-1 (<bold>B</bold>) and normal dental pulp collagen fibrils components COL-I (<bold>J</bold>), COL-III (<bold>K</bold>) showed weakly positive expression. However, most cells were stained negatively for TGF-β and Fibronectin Scale bars = 50 μm.</p></caption><graphic xlink:href="srep44635-f5"></graphic></fig><fig id="f6"><label>Figure 6</label><caption><title>Formation of new tissue.</title><p>(<bold>A</bold>) The bone like tissue were formed on the pulp cavity surface of TDM. Many osseous lacunas spreaded over newly-formed bone matrix (<bold>B</bold>). We could also observe the osteoblast like cells at the edge of bone matrix (The yellow arrows had pointed out) (<bold>C</bold>). In Masson staining, newly-formed bone like tissue were stained red instead of blue suggested that the tissue contained less collagen components (<bold>D</bold>). (The yellow spots labeled the layer of newly-formed bone-like tissue, TDM: treated dentin matrix, NB: newly-formed bone-like tissue). Scale bars = 200 μm for A. Scale bars = 100 μm for (<bold>B</bold>,<bold>C</bold>). Scale bars = 50 μm for (<bold>D</bold>).</p></caption><graphic xlink:href="srep44635-f6"></graphic></fig></floats-group></pmc><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Luo, Shiwei" sort="Luo, Shiwei" uniqKey="Luo S" first="Shiwei" last="Luo">Shiwei Luo</name></noRegion><name sortKey="Guo, Weihua" sort="Guo, Weihua" uniqKey="Guo W" first="Weihua" last="Guo">Weihua Guo</name><name sortKey="Guo, Weihua" sort="Guo, Weihua" uniqKey="Guo W" first="Weihua" last="Guo">Weihua Guo</name><name sortKey="Guo, Weihua" sort="Guo, Weihua" uniqKey="Guo W" first="Weihua" last="Guo">Weihua Guo</name><name sortKey="He, Wei" sort="He, Wei" uniqKey="He W" first="Wei" last="He">Wei He</name><name sortKey="Li, Rui" sort="Li, Rui" uniqKey="Li R" first="Rui" last="Li">Rui Li</name><name sortKey="Luo, Shiwei" sort="Luo, Shiwei" uniqKey="Luo S" first="Shiwei" last="Luo">Shiwei Luo</name><name sortKey="Luo, Shiwei" sort="Luo, Shiwei" uniqKey="Luo S" first="Shiwei" last="Luo">Shiwei Luo</name><name sortKey="Pei, Fei" sort="Pei, Fei" uniqKey="Pei F" first="Fei" last="Pei">Fei Pei</name><name sortKey="Pei, Fei" sort="Pei, Fei" uniqKey="Pei F" first="Fei" last="Pei">Fei Pei</name><name sortKey="Pei, Fei" sort="Pei, Fei" uniqKey="Pei F" first="Fei" last="Pei">Fei Pei</name><name sortKey="Tian, Weidong" sort="Tian, Weidong" uniqKey="Tian W" first="Weidong" last="Tian">Weidong Tian</name><name sortKey="Tian, Weidong" sort="Tian, Weidong" uniqKey="Tian W" first="Weidong" last="Tian">Weidong Tian</name><name sortKey="Tian, Weidong" sort="Tian, Weidong" uniqKey="Tian W" first="Weidong" last="Tian">Weidong Tian</name><name sortKey="Zhang, Wen" sort="Zhang, Wen" uniqKey="Zhang W" first="Wen" last="Zhang">Wen Zhang</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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