title: Cell Biological Assays for Measuring Odontogenic Activities of CCN Proteins.
abstract: In postnatal dentin formation, odontoblast differentiation occurs in the pulp tissue regenerative process under pathological condition. Odontoblasts and newly differentiated odontoblast-like cells beneath the caries lesion form tertiary dentin and are highly odontogenic. To observe the activity of dentinogenesis occur within the hard tissue, a combination of immunohistological analysis and immunodetection of dentinogenesis specific molecules, such as dentin sialophosphoprotein (DSPP) and/or its cleaved products dentin sialoprotein (DSP) and dentin phosphoprotein (DPP), is a reliable approach. Besides, recent studies have revealed that the expression of CCN family member 2 (CCN2), a member of the CCN family protein, is confirmed in accordance with tooth development and reparative dentin formation. Therefore, CCN2 could serve as a marker for dentinogenesis. Here, we describe a method for visualizing the CCN2 signal as an odontogenic activity in formalin-fixed paraffin-embedded (FFPE) sections of demineralized human teeth and human dental pulp cells.
authors.affiliation: Department of Pulp Biology and Endodontics, Graduate School of Dentistry, Kanagawa Dental University, Yokosuka, Kanagawa, 238-8580, Japan. muromachi@kdu.ac.jp.
authors.forename: Koichiro
authors.initials: K
authors.lastname: Muromachi
authors.affiliation: Laboratory of Veterinary Biochemistry, Nihon University College of Bioresource Sciences, Fujisawa, Kanagawa, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sugiya
authors.affiliation: Department of Pulp Biology and Endodontics, Graduate School of Dentistry, Kanagawa Dental University, Yokosuka, Kanagawa, 238-8580, Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Tani-Ishii
mesh_terms: D001681:Biological Assay; D015153:Blotting, Western; D055511:CCN Intercellular Signaling Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D002478:Cells, Cultured; D055513:Connective Tissue Growth Factor / Q000235:genetics / Q000378:metabolism; D003782:Dental Pulp / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D009804:Odontoblasts / Q000187:drug effects / Q000378:metabolism; D009805:Odontogenesis / Q000187:drug effects* / Q000235:genetics; D009195:Peroxidase / Q000378:metabolism
chemical_list: D055511:CCN Intercellular Signaling Proteins; C526317:CCN2 protein, human; D055513:Connective Tissue Growth Factor; D009195:Peroxidase
keywords: Dentinogenesis; Dentin–pulp complex; Human teeth; Immunohistochemistry; Odontoblasts; Western blot