Galectin-3 Plays an Important Role in BMP7-Induced Cementoblastic Differentiation of Human Periodontal Ligament Cells by Interacting with Extracellular Components

Figure 1

Anti-CM3 antibody binds to BMP7-induced cementoblast-like cells and cementum in tooth root. (a) Cytodifferentiation of hPDLCs into cementoblast-like cells via BMP7 treatment together with SB431542, an inhibitor of TGF-β1 signaling. Cementoblast-like cells were identified via increased transcriptional expression of cementoblast-specific markers such as CAP (A), CEMP1 (B), OSX (C), OCN (D), and BSP (E). Reversely, PDL fibroblastic markers such as SCX (F) and PLAP-1 (G) were upregulated in hPDLCs treated with TGF-β1. Statistical significance is determined using Student’s -test (). ; ; . (b) Cell-binding affinity of anti-CM3 antibody using FACS analysis. Cells were detached from the culture dish under the nonenzymatic condition, and anti-CM3 antibody was added to the intact cells, followed by FITC-labeled secondary antibody. Human periodontal ligament stem cells (A) and mouse cell lines (B) were used for the antibody-binding experiment. (c) Western blot analysis of endogenous CM3 antigen in hPDLCs (A) and mouse cell lines (B). In (b, c), TGF-β1 and B7/SB indicate hPDLCs treated with 10 ng/mL TGF-β1 and hPDLCs treated with 100 ng/mL BMP7 and 10 μM SB431542, respectively. OCCM and NIH3T3 indicate mouse cementoblast cells and mouse fibroblast cells, respectively. (d) Immunohistochemistry via anti-CM3 antibody. Antibodies were accumulated in the cementum of the tooth. (A) No staining control without antibody; (B) with anti-CM3 antibody; (C) with anti-OSX antibody as a positive control for the cementum; (D) with anti-PLAP-1 antibody as a negative control. AB: alveolar bone; PDL: periodontal ligament; CM: cementum; DT: dentin.

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