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Study on site-specific expression of bone formation and resorption factors in human dental follicles

Journal article
Authors Pamela Uribe
P. Plakwicz
Lena Larsson
E. Czochrowska
Anna Westerlund
Maria Ransjö
Published in European Journal of Oral Sciences
Volume 126
Issue 6
Pages 439-448
ISSN 0909-8836
Publication year 2018
Published at Institute of Odontology
Institute of Odontology, Section 3
Pages 439-448
Language en
Links dx.doi.org/10.1111/eos.12568
Keywords connexin 43, osteoblast, osteoclast, receptor activator of nuclear factor-kappa B ligand, tooth eruption, gene-expression, alveolar-bone, osteoblast, differentiation, in-vitro, rankl, cells, osteoclastogenesis, pcr, osteoprotegerin, Dentistry, Oral Surgery & Medicine
Subject categories Dentistry

Abstract

We sought to investigate site-specific expression of bone-regulatory factors expressed by human dental follicles and to compare the stimulated expression of tumour necrosis factor (ligand) superfamily, member 11/tumour necrosis factor receptor superfamily, member 11b (RANKL/OPG) in human dental follicle cells (HDFCs) from different patients. Analysis of bone-regulatory markers in follicles from 12 different study participants was performed using RT-qPCR and immunofluorescence; apical and coronal segments from each dental follicle were processed independently. Four additional dental follicles were used for cell cultures; HDFCs were precultured in osteogenic medium to initiate differentiation and thereafter cultured with 10(-6) M forskolin (FSK) to activate the protein kinase cAMP (PKA/cAMP) signalling pathway and induce RANKL/OPG expression. We demonstrate that RANKL expression is significantly higher in the coronal part of follicles than in the apical part. High levels of collagen type 1 (COL1), alkaline phosphatase (ALP) and Gap-junction protein, alpha 1, 43 kDa (CX43) were expressed, whereas expression of Sp7 transcription factor (OSX), bone morphogenetic protein 2 (BMP2), colony-stimulating factor 1 (CSF-1), chemokine (C-C motif) ligand 2 (MCP1), and OPG was low in all samples. The immunofluorescence localization of CSF-1, MCP1, osteocalcin (OCN), RANKL, and BMP2 was not specific for either part of the follicles. In conclusion, a consistently high expression of CX43 suggests that gap-junction communication in HDFCs is essential for the eruption process. Furthermore, the induced expression of RANKL in HDFCs varies significantly between individuals and may relate to clinical variations in tooth eruption.

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