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Gingival fibroblasts prevent BMP‐mediated osteoblastic differentiation

OBJECTIVES: The inhibitory action of the superficial gingival connective tissues may limit the regenerative potential of alveolar bone in periodontal therapy or dental implant applications. The aims of this study were to investigate the hypothesis that gingival fibroblasts (GF) can inhibit bone morp...

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Autores principales: Ghuman, Mandeep S., Al‐Masri, Maher, Xavier, Guilherme, Cobourne, Martyn T., McKay, Ian J., Hughes, Francis J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6492095/
https://www.ncbi.nlm.nih.gov/pubmed/30511378
http://dx.doi.org/10.1111/jre.12631
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author Ghuman, Mandeep S.
Al‐Masri, Maher
Xavier, Guilherme
Cobourne, Martyn T.
McKay, Ian J.
Hughes, Francis J.
author_facet Ghuman, Mandeep S.
Al‐Masri, Maher
Xavier, Guilherme
Cobourne, Martyn T.
McKay, Ian J.
Hughes, Francis J.
author_sort Ghuman, Mandeep S.
collection PubMed
description OBJECTIVES: The inhibitory action of the superficial gingival connective tissues may limit the regenerative potential of alveolar bone in periodontal therapy or dental implant applications. The aims of this study were to investigate the hypothesis that gingival fibroblasts (GF) can inhibit bone morphogenetic protein (BMP)‐induced osteoblastic differentiation, to determine their expression of BMP inhibitors, and finally to determine whether reduction of these inhibitors can relieve suppression of osteoblastic differentiation. METHODS: Gingival fibroblasts were co‐cultured either directly or indirectly with calvarial osteoblasts to assess alkaline phosphatase inhibitory activity, a marker of osteoblastic differentiation. To test total BMP‐inhibitory activity of rat GF, conditioned media (GFCM) were collected from cultures. ROS 17/2.8 osteoblastic cells were stimulated with BMP2, together with GFCM. Inhibitor expression was tested using RT‐qPCR, Western blotting and in situ hybridization. Removal of inhibitors was carried out using immunoprecipitation beads. RESULTS: Co‐culture experiments showed GF‐secreted factors that inhibit BMP‐stimulated ALP activity. 10 ng/ml BMP2 increased alkaline phosphatase expression in ROS cells by 41%. GFCM blocked BMP activity which was equivalent to the activity of 100 ng/ml Noggin, a well‐described BMP inhibitor. Cultured gingival fibroblasts constitutively expressed BMP antagonist genes from the same subfamily, Grem1, Grem2 and Nbl1 and the Wnt inhibitor Sfrp1. Gremlin1 (6.7 × reference gene expression) had highest levels of basal expression. ISH analysis showed Gremlin1 expression was restricted to the inner half of the gingival lamina propria and the PDL. Removal of Gremlin1 protein from GFCM eliminated the inhibitory effect of GFCM on ALP activity in ROS cells. Subsequent addition of recombinant Gremlin1 restored the inhibitory activity. CONCLUSIONS: Factors secreted by gingival fibroblasts inhibit BMP‐induced bone formation and a range of BMP inhibitors are constitutively expressed in gingival connective tissues. These inhibitors, particularly Gremlin1, may limit coronal alveolar bone regenerative potential during oral and periodontal surgery.
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spelling pubmed-64920952019-05-06 Gingival fibroblasts prevent BMP‐mediated osteoblastic differentiation Ghuman, Mandeep S. Al‐Masri, Maher Xavier, Guilherme Cobourne, Martyn T. McKay, Ian J. Hughes, Francis J. J Periodontal Res Original Articles OBJECTIVES: The inhibitory action of the superficial gingival connective tissues may limit the regenerative potential of alveolar bone in periodontal therapy or dental implant applications. The aims of this study were to investigate the hypothesis that gingival fibroblasts (GF) can inhibit bone morphogenetic protein (BMP)‐induced osteoblastic differentiation, to determine their expression of BMP inhibitors, and finally to determine whether reduction of these inhibitors can relieve suppression of osteoblastic differentiation. METHODS: Gingival fibroblasts were co‐cultured either directly or indirectly with calvarial osteoblasts to assess alkaline phosphatase inhibitory activity, a marker of osteoblastic differentiation. To test total BMP‐inhibitory activity of rat GF, conditioned media (GFCM) were collected from cultures. ROS 17/2.8 osteoblastic cells were stimulated with BMP2, together with GFCM. Inhibitor expression was tested using RT‐qPCR, Western blotting and in situ hybridization. Removal of inhibitors was carried out using immunoprecipitation beads. RESULTS: Co‐culture experiments showed GF‐secreted factors that inhibit BMP‐stimulated ALP activity. 10 ng/ml BMP2 increased alkaline phosphatase expression in ROS cells by 41%. GFCM blocked BMP activity which was equivalent to the activity of 100 ng/ml Noggin, a well‐described BMP inhibitor. Cultured gingival fibroblasts constitutively expressed BMP antagonist genes from the same subfamily, Grem1, Grem2 and Nbl1 and the Wnt inhibitor Sfrp1. Gremlin1 (6.7 × reference gene expression) had highest levels of basal expression. ISH analysis showed Gremlin1 expression was restricted to the inner half of the gingival lamina propria and the PDL. Removal of Gremlin1 protein from GFCM eliminated the inhibitory effect of GFCM on ALP activity in ROS cells. Subsequent addition of recombinant Gremlin1 restored the inhibitory activity. CONCLUSIONS: Factors secreted by gingival fibroblasts inhibit BMP‐induced bone formation and a range of BMP inhibitors are constitutively expressed in gingival connective tissues. These inhibitors, particularly Gremlin1, may limit coronal alveolar bone regenerative potential during oral and periodontal surgery. John Wiley and Sons Inc. 2018-12-03 2019-06 /pmc/articles/PMC6492095/ /pubmed/30511378 http://dx.doi.org/10.1111/jre.12631 Text en © 2018 The Authors. Journal of Periodontal Research Published by John Wiley & Sons Ltd This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Ghuman, Mandeep S.
Al‐Masri, Maher
Xavier, Guilherme
Cobourne, Martyn T.
McKay, Ian J.
Hughes, Francis J.
Gingival fibroblasts prevent BMP‐mediated osteoblastic differentiation
title Gingival fibroblasts prevent BMP‐mediated osteoblastic differentiation
title_full Gingival fibroblasts prevent BMP‐mediated osteoblastic differentiation
title_fullStr Gingival fibroblasts prevent BMP‐mediated osteoblastic differentiation
title_full_unstemmed Gingival fibroblasts prevent BMP‐mediated osteoblastic differentiation
title_short Gingival fibroblasts prevent BMP‐mediated osteoblastic differentiation
title_sort gingival fibroblasts prevent bmp‐mediated osteoblastic differentiation
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6492095/
https://www.ncbi.nlm.nih.gov/pubmed/30511378
http://dx.doi.org/10.1111/jre.12631
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