Resumen de Enamel Matrix Derivative Induces Connective Tissue Growth Factor Expression in Human Osteoblastic Cells

Jean-Pierre Bernimoulin, Nicole Pischon, Nora Heng, Philippe D. N'Guessan, Bernd-Michael Kleber

• Abstract Journal of Periodontology December 2007, Vol. 78, No. 12, Pages 2369-2379 , DOI 10.1902/jop.2007.070130 (doi:10.1902/jop.2007.070130) Enamel Matrix Derivative Induces Connective Tissue Growth Factor Expression in Human Osteoblastic Cells Nora H.M. Heng,* Philippe D. N'Guessan,† Bernd-Michael Kleber,* Jean-Pierre Bernimoulin,* and Nicole Pischon* *Institute of Periodontology and Synoptic Dentistry, Charité-Medical University of Berlin, Berlin, Germany.

  †Department of Internal Medicine/Infectious Diseases and Respiratory Medicine, Charité-Medical University of Berlin.

  Correspondence: Dr. Nicole Pischon, Institute of Periodontology and Synoptic Dentistry, Charité-Medical University of Berlin, Campus Virchow Klinikum, Augustenburger Platz 1, 13353 Berlin, Germany. Fax: 49-30-450-562-931; e-mail: nicole.pischon@charite.de.

  Background: Enamel matrix derivative (EMD) stimulates the production of transforming growth factor-beta (TGF-β), which has been suggested to play a role in mediating the effects of EMD in periodontal tissue regeneration. Connective tissue growth factor (CTGF) is a mediator of TGF-β and promotes cell development. The interaction between EMD and CTGF is unknown. This study explored the effects of EMD on CTGF expression in human osteoblastic cells and whether the interaction is modulated by the TGF-β signaling pathway. Also, the roles of CTGF in cell proliferation, cell cycle progression, and mineralized nodule formation of EMD-induced osteoblastic cultures were examined.

  Methods: Human osteoblastic cells (Saos-2) were treated with 25 to 100 μg/ml EMD with or without the addition of TGF-β inhibitor. CTGF mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR), and CTGF protein levels were assayed by Western blot analysis. In addition, cell cycle progression and DNA synthesis were determined by flow cytometry and 5-bromo-2′-deoxyuridine (BrdU) incorporation following EMD treatment with or without CTGF antibody. Mineralization was examined by alizarin red staining and quantified by elution with cetylpyridinium chloride.

  Results: Western blot and RT-PCR analysis demonstrated a dose-dependent increase of CTGF expression by EMD. EMD-induced CTGF expression was reduced significantly in the presence of TGF-β inhibitor. Cell cycle and BrdU analysis revealed an increase in cell proliferation following EMD treatment, which was due to an increase in the percentage of cells in the G2/M phase of the cell cycle. No significant effect was found when anti-CTGF antibody was added. Conversely, mineralization was inhibited significantly in EMD-treated cultures in the presence of anti-CTGF antibody.

  Conclusions: EMD stimulates CTGF expression, and the interaction is modulated via TGF-β in osteoblastic cells. Also, CTGF affects EMD-induced osteoblastic mineralization but not cell proliferation. To our knowledge, these results provide novel insight into EMD–CTGF interaction, two biomodifiers that have therapeutic relevance to tissue engineering and regeneration.

  KEYWORDS: Connective tissue growth factor, osteoblasts, periodontal regeneration, transforming growth factor-beta Cited by Nora H.M. Heng, Janine Zahlten, Valerie Cordes, Marianne M-A Ong, Bee Tin Goh, Philippe D. N’Guessan and Nicole Pischon. (2015) Effects of Enamel Matrix Derivative and Transforming Growth Factor-β1 on Connective Tissue Growth Factor in Human Periodontal Ligament Fibroblasts. Journal of Periodontology 86:4, 569-577.

  Online publication date: 1-Apr-2015.

  Abstract | Full Text | PDF (786 KB) | PDF Plus (643 KB) R. Gruber, A. Stähli, R. J. Miron, D. D. Bosshardt and A. Sculean. (2015) Common target genes of palatal and gingival fibroblasts for EMD: the microarray approach. Journal of Periodontal Research 50:10.1111/jre.2014.50.issue-1, 103-112.

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