Premature Osteoblast Clustering by Enamel Matrix Proteins Induces Osteoblast Differentiation through Up-Regulation of Connexin 43 and N-Cadherin

Miron, Richard J.; Hedbom, Erik; Ruggiero, Sabrina; Bosshardt, Dieter D.; Zhang, Yufeng; Mauth, Corinna; Gemperli, Anja C.; Iizuka, Tetsuya; Buser, Daniel; Sculean, Anton

doi:10.1371/journal.pone.0023375

Cited by 42 publications

(37 citation statements)

References 69 publications

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“…In this study, a time-dependant reduction in osteoblast differentiation was observed when cells were left unpassaged from 7 to 28 days following treatment with EMD. Interestingly, we have previously reported that EMD enhances osteoblast differentiation via cell-cell contacts by upregulating osteoblast gap junctional proteins, connexin 43 ( cx43 ) [32]. Gap junctions are aqueous transmembrane channels that connect the cytoplasm of two adjacent cells and allow the diffusion of small molecules with a molecular mass of less than 1 kDa such as small metabolites, ions, and intracellular signaling molecules (calcium, cAMP, and inositol triphosphate) to pass through [57].…”

Section: Discussionmentioning

confidence: 99%

“…Gap junctions are aqueous transmembrane channels that connect the cytoplasm of two adjacent cells and allow the diffusion of small molecules with a molecular mass of less than 1 kDa such as small metabolites, ions, and intracellular signaling molecules (calcium, cAMP, and inositol triphosphate) to pass through [57]. Cx43 molecules are absolutely essential for osteoblast differentiation and function [58]–[61] and EMD targets their upregulation [32]. The results from the present study may also indicate that cx43 may be implicated in the down-regulation of osteoblast differentiation in response to EMD following cell maturation.…”

Section: Discussionmentioning

confidence: 99%

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Influence of Enamel Matrix Derivative on Cells at Different Maturation Stages of Differentiation

et al. 2013

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Enamel matrix derivative (EMD), a porcine extract harvested from developing porcine teeth, has been shown to promote formation of new cementum, periodontal ligament and alveolar bone. Despite its widespread use, an incredibly large variability among in vitro studies has been observed. The aim of the present study was to determine the influence of EMD on cells at different maturation stages of osteoblast differentiation by testing 6 cell types to determine if cell phenotype plays a role in cell behaviour following treatment with EMD. Six cell types including MC3T3-E1 pre-osteoblasts, rat calvarial osteoblasts, human periodontal ligament (PDL) cells, ROS cells, MG63 cells and human alveolar osteoblasts were cultured in the presence or absence of EMD and proliferation rates were quantified by an MTS assay. Gene expression of collagen1(COL1), alkaline phosphate(ALP) and osteocalcin(OC) were investigated by real-time PCR. While EMD significantly increased cell proliferation of all cell types, its effect on osteoblast differentiation was more variable. EMD significantly up-regulated gene expression of COL1, ALP and OC in cells early in their differentiation process when compared to osteoblasts at later stages of maturation. Furthermore, the effect of cell passaging of primary human PDL cells (passage 2 to 15) was tested in response to treatment with EMD. EMD significantly increased cell proliferation and differentiation of cells at passages 2–5 however had completely lost their ability to respond to EMD by passages 10+. The results from the present study suggest that cell stimulation with EMD has a more pronounced effect on cells earlier in their differentiation process and may partially explain why treatment with EMD primarily favors regeneration of periodontal defects (where the periodontal ligament contains a higher number of undifferentiated progenitor cells) over regeneration of pure alveolar bone defects containing no periodontal ligament and a more limited number of osteoprogenitor cells.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Influence of Enamel Matrix Derivative on Cells at Different Maturation Stages of Differentiation

et al. 2013

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“…For counting the cell number, 4′,6-diamidino-2-phenylindole (DAPI) was applied to visualize the nuclei as previously described [48]. At each time point, the Ti structures were washed with PBS to remove non-attached cells and fixed in 4% formaldehyde for 10 min followed by staining with DAPI.…”

Section: Methodsmentioning

confidence: 99%

Behavior of Gingival Fibroblasts on Titanium Implant Surfaces in Combination with either Injectable-PRF or PRP

Wang

Zhang

Choukroun³

et al. 2017

IJMS

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Various strategies have been employed to speed tissue regeneration using bioactive molecules. Interestingly, platelet concentrates derived from a patient’s own blood have been utilized as a regenerative strategy in recent years. In the present study, a novel liquid platelet formulation prepared without the use of anti-coagulants (injectable-platelet-rich fibrin, i-PRF) was compared to standard platelet-rich plasma (PRP) with gingival fibroblasts cultured on smooth and roughened titanium implant surfaces. Standard PRP and i-PRF (centrifuged at 700 rpm (60× g) for 3 min) were compared by assays for fibroblast biocompatibility, migration, adhesion, proliferation, as well as expression of platelet-derived growth factor (PDGF), transforming growth factor-β (TGF-β), collagen1 (COL1) and fibronectin (FN). The results demonstrate that i-PRF induced significantly higher cell migration, as well as higher messenger RNA (mRNA) levels of PDGF, TGF-β, collagen1 and fibronectin when compared to PRP. Furthermore, collagen1 synthesis was highest in the i-PRF group. These findings demonstrate that liquid platelet concentrates can be formulated without the use of anticoagulants and present much translational potential for future research. Future animal and clinical trials are now necessary to further investigate the potential of utilizing i-PRF for soft tissue regenerative protocols in combination with various biomaterials.

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“…A number of studies have demonstrated the osteoconductive activity of EMD, in particular, for human periodontal ligament cells and osteoblastic cell types [13]. EMD enhances osteoblast differentiation and mineralization [14], as well as contributes to multi-lineage differentiation of human periodontal ligament cells [15]. Furthermore, transplantation of induced pluripotent stem cells combined with EMD greatly enhanced periodontal tissue regeneration [16].…”

Section: Introductionmentioning

confidence: 99%

Identification of Novel Amelogenin-Binding Proteins by Proteomics Analysis

et al. 2013

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Emdogain (enamel matrix derivative, EMD) is well recognized in periodontology. It is used in periodontal surgery to regenerate cementum, periodontal ligament, and alveolar bone. However, the precise molecular mechanisms underlying periodontal regeneration are still unclear. In this study, we investigated the proteins bound to amelogenin, which are suggested to play a pivotal role in promoting periodontal tissue regeneration. To identify new molecules that interact with amelogenin and are involved in osteoblast activation, we employed coupling affinity chromatography with proteomic analysis in fractionated SaOS-2 osteoblastic cell lysate. In SaOS-2 cells, many of the amelogenin-interacting proteins in the cytoplasm were mainly cytoskeletal proteins and several chaperone molecules of heat shock protein 70 (HSP70) family. On the other hand, the proteomic profiles of amelogenin-interacting proteins in the membrane fraction of the cell extracts were quite different from those of the cytosolic-fraction. They were mainly endoplasmic reticulum (ER)-associated proteins, with lesser quantities of mitochondrial proteins and nucleoprotein. Among the identified amelogenin-interacting proteins, we validated the biological interaction of amelogenin with glucose-regulated protein 78 (Grp78/Bip), which was identified in both cytosolic and membrane-enriched fractions. Confocal co-localization experiment strongly suggested that Grp78/Bip could be an amelogenin receptor candidate. Further biological evaluations were examined by Grp78/Bip knockdown analysis with and without amelogenin. Within the limits of the present study, the interaction of amelogenin with Grp78/Bip contributed to cell proliferation, rather than correlate with the osteogenic differentiation in SaOS-2 cells. Although the biological significance of other interactions are not yet explored, these findings suggest that the differential effects of amelogenin-derived osteoblast activation could be of potential clinical significance for understanding the cellular and molecular bases of amelogenin-induced periodontal tissue regeneration.

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Premature Osteoblast Clustering by Enamel Matrix Proteins Induces Osteoblast Differentiation through Up-Regulation of Connexin 43 and N-Cadherin

Cited by 42 publications

References 69 publications

Influence of Enamel Matrix Derivative on Cells at Different Maturation Stages of Differentiation

Influence of Enamel Matrix Derivative on Cells at Different Maturation Stages of Differentiation

Behavior of Gingival Fibroblasts on Titanium Implant Surfaces in Combination with either Injectable-PRF or PRP

Identification of Novel Amelogenin-Binding Proteins by Proteomics Analysis

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