Functional characterization of the parathyroid hormone 1 receptor in human periodontal ligament cells

Abuduwali, Nuersailike; Lossdörfer, Stefan; Winter, Jochen; Kraus, Dominik; Guhlke, Stefan; Wolf, Michael; Jäger, Andreas

doi:10.1007/s00784-013-0985-4

Cited by 12 publications

(11 citation statements)

References 56 publications

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“…To demonstrate the effect of HMGB1 on early and late osteoblastic differentiation of hPDL cells, immunocytochemical staining on cover was performed using either a 1 : 50 (40 μ g/mL) working dilution of the primary antibody for ALP (rabbit anti-human; Quartett, Germany) or 1 : 25 (80 μ g/mL) for osteocalcin (rabbit anti-human; Biozol, Germany) in combination with a secondary fluorescence-labeled (Cy-3) antibody (1 : 100 (1.5 μ g/mL) working dilution; goat anti-rabbit; Invitrogen, Germany) and analyzed microscopically as reported previously [ 24 ]. Negative control were run by (i) omitting the primary antibody, (ii) omitting the primary and secondary antibody and using TBS/BSA instead, and (iii) by substituting the primary antibody by an unspecific IgG.…”

Section: Methodsmentioning

confidence: 99%

Anabolic Properties of High Mobility Group Box Protein-1 in Human Periodontal Ligament CellsIn Vitro

Wolf

Lossdörfer

Römer

et al. 2014

Mediators of Inflammation

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High mobility group box protein-1 (HMGB1) is mainly recognized as a chemoattractant for macrophages in the initial phase of host response to pathogenic stimuli. However, recent findings provide evidence for anabolic properties in terms of enhanced proliferation, migration, and support of wound healing capacity of mesenchymal cells suggesting a dual role of the cytokine in the regulation of immune response and subsequent regenerative processes. Here, we examined potential anabolic effects of HMGB1 on human periodontal ligament (PDL) cells in the regulation of periodontal remodelling, for example, during orthodontic tooth movement. Preconfluent human PDL cells (hPDL) were exposed to HMGB1 protein and the influence on proliferation, migration, osteogenic differentiation, and biomineralization was determined by MTS assay, real time PCR, immunofluorescence cytochemistry, ELISA, and von Kossa staining. HMGB1 protein increased hPDL cell proliferation, migration, osteoblastic marker gene expression, and protein production as well as mineralized nodule formation significantly. The present findings support the dual character of HMGB1 with anabolic therapeutic potential that might support the reestablishment of the structural and functional integrity of the periodontium following periodontal trauma such as orthodontic tooth movement.

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Section: Methodsmentioning

confidence: 99%

Anabolic Properties of High Mobility Group Box Protein-1 in Human Periodontal Ligament CellsIn Vitro

Wolf

Lossdörfer

Römer

et al. 2014

Mediators of Inflammation

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“…PTH1R is recognized by both PTH‐ and PTH‐related peptide (PTHrP) . PTH1R is coupled to the Gsα/PKA and the Gq/PLC/PKC . Previous studies have suggested that the activation of renal 1,25(OH) 2 D synthesis by PTH is via activating PKA signalling pathway and the regulation of renal 1,25(OH) 2 D secretion is via activating PKC signalling pathway .…”

Section: Introductionmentioning

confidence: 99%

Regulation of 25‐hydroxyvitamin D‐1‐hydroxylase and 24‐hydroxylase in keratinocytes by PTH and FGF23

Fan

Jiang

et al. 2018

Experimental Dermatology

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Renal 25-hydroxyvitamin D-1α-hydroxylase (1αOHase, CYP27B1) and 24-hydroxylase (24OHase, CYP24A1) are tightly regulated. However, little is known about the regulation of 1α(OH)ase and 24(OH)ase in extrarenal tissue such as the epidermis. This study was to determine the roles of parathyroid hormone (PTH) and fibroblast growth factor 23 (FGF 23) in the regulation of 1α(OH)ase and 24(OH)ase in epidermal keratinocytes as well as epidermal keratinocyte proliferation and differentiation. The results showed that PTH increased the protein level of 1α(OH)ase in human epidermal keratinocyte cell line HaCaT, but had no effect on the level of 24(OH)ase. The effect of PTH on 1α(OH)ase was blocked by the PKC inhibitor. Treatment with FGF23 decreased mRNA and protein levels of 1α(OH)ase and increased mRNA and protein levels of 24(OH)ase in HaCaT cells. The effect of FGF23 on 1α(OH)ase and 24(OH)ase was blocked by the mitogen-activated protein kinase/extracellular regulated protein kinase (MAPK/ERK) inhibitor. In addition, treatment with PTH enhanced levels of differentiation markers including keratin 1, involucrin, loricrin, and filaggrin but reduced levels of BrdU incorporation in HaCaT cells. These effects were inhibited by the PKC inhibitor. FGF23 enhanced proliferation of HaCaT cells, but reduced levels of early differentiation markers including keratin 1 and involucrin and enhanced levels of the later differentiation markers including loricrin and filaggrin. These results suggest that PTH stimulates 1α(OH)ase expression and differentiation of HaCaT cells and inhibits proliferation via PKC. The data also suggest that FGF23 inhibits 1α(OH)ase expression and stimulates 24(OH)ase expression via MAPK/ERK. In addition, FGF23 enhances proliferation and late differentiation and inhibits early differentiation of HaCaT keratinocytes.

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“…30,31 The latter have been reported to share osteoblast-like traits and respond accordingly to hormonal stimulation. 32,33 PDL cells were shown to express the PTH 1 receptor (1R) 29 and respond to anabolic doses of PTH with changes of many crucial cell functions including proliferation, differentiation, apoptosis, and the production of key molecules of bone remodeling. [15][16][17] These findings are in line with the data obtained here in the monoculture experiments regarding proliferation and differentiation and strengthen the view of PDL cells as important regulators of periodontal remodeling.…”

Section: Discussionmentioning

confidence: 99%

“…To visualize the effect of Wnt10b on hPDL cell differentiation along the osteoblastic pathway, immunocytochemical staining was performed using either a 1:50 (40 mg/ml) working dilution of the primary Ab for ALP (rabbit anti-human; Quartett, Germany) or 1:25 (80 mg/ml) for osteocalcin (rabbit anti-human; Biozol, Germany) in combination with a secondary fluorescence-labeled (Cy-3) Ab (1:100 (1.5 mg/ml) working dilution; goat anti-rabbit; Invitrogen, Germany) and analyzed microscopically as reported previously. 29 Negative controls were run by (i) omitting the primary Ab, (ii) omitting the primary and secondary Ab and using TBS/BSA instead, and (iii) by substituting the primary Ab by an unspecific IgG. The same protocol was followed for detection of LRP5 in hPDL cells using a monoclonal rabbit anti-human Ab (Cell Signaling Technology Europe, The Netherlands) using a 1:100 (20 mg/ml) working dilution.…”

Section: Immunofluorescence Cytochemistrymentioning

confidence: 99%

CD8+ T cells mediate the regenerative PTH effect in hPDL cells via Wnt10b signaling

et al. 2016

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It was the aim of the present investigation to examine whether the stimulating effect of parathyroid hormone (PTH) on human periodontal ligament (hPDL) cell proliferation and differentiation would be enhanced by hPDL/T-cell interaction involving Wnt10b signaling as a mediating pathway. hPDL cells were cultured from healthy premolar tissues of three adolescent orthodontic patients and exposed to PTH(1-34) in monocultures or co-cultures with CD8 T cells. At harvest, proliferation, alkaline phosphatase-specific activity (ALP), and osteocalcin production were determined by immunofluorescence cytochemistry, real-time PCR, biochemical assay, and ELISA. Wnt10b signaling was analyzed by the use of a specific WNT10b neutralizing antibody. PTH(1-34) stimulation of T cells significantly increased Wnt10b expression and production. Wnt10b exposure of hPDL cells enhanced proliferation and differentiation. PDL cells co-cultured with T cells showed a Wnt10b-dependent regulation of proliferation and differentiation parameters. The addition of a Wnt10b-neutralizing Ab to the co-culture medium resulted in a significant inhibition of the PTH(1-34) effect on proliferation, ALP-specific activity, and osteocalcin protein expression. Our findings provide novel insight into the mechanism of action of PTH on hPDL cells and establish the interplay of T cells and hPDL cells via the Wnt10b pathway as a modulating factor for the anabolic properties of the hormone in periodontal regeneration.

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Functional characterization of the parathyroid hormone 1 receptor in human periodontal ligament cells

Abstract: The findings further elucidate the characteristics of PTH action in dental tissues and widen the theoretical basis for the development of anabolic treatment strategies.

Cited by 12 publications

References 56 publications

Anabolic Properties of High Mobility Group Box Protein-1 in Human Periodontal Ligament CellsIn Vitro

Anabolic Properties of High Mobility Group Box Protein-1 in Human Periodontal Ligament CellsIn Vitro

Regulation of 25‐hydroxyvitamin D‐1‐hydroxylase and 24‐hydroxylase in keratinocytes by PTH and FGF23

CD8+ T cells mediate the regenerative PTH effect in hPDL cells via Wnt10b signaling

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