Zirconia stimulates ECM-remodeling as a prerequisite to pre-osteoblast adhesion/proliferation by possible interference with cellular anchorage
- 281 Downloads
The biological response to zirconia (ZrO2) is not completely understood, which prompted us to address its effect on pre-osteoblastic cells in both direct and indirect manner. Our results showed that zirconia triggers important intracellular signaling mainly by governing survival signals which leads to cell adhesion and proliferation by modulating signaling cascade responsible for dynamic cytoskeleton rearrangement, as observed by fluorescence microscopy. The phosphorylations of Focal Adhesion Kinase (FAK) and Rac1 decreased in response to ZrO2 enriched medium. This corroborates the result of the crystal violet assay, which indicated a significant decrease of pre-osteoblast adhesion in responding to ZrO2 enriched medium. However, we credit this decrease on pre-osteoblast adhesion to the need to govern intracellular repertory of intracellular pathways involved with cell cycle progression, because we found a significant up-phosphorylation of Mitogen-Activated Protein Kinase (MAPK)-p38 and Cyclin-dependent kinase 2 (CDK2), while p15 (a cell cycle suppressor) decreased. Importantly, Protein phosphatase 2 A (PP2A) activity decreased, guaranteeing the significant up-phosphorylation of MAPK -p38 in response to ZrO2 enriched medium. Complementarily, there was a regulation of Matrix Metalloproteinases (MMPs) in response to Zirconia and this remodeling could affect cell phenotype by interfering on cell anchorage. Altogether, our results show a repertory of signaling molecules, which suggests that ECM remodel as a pre-requisite to pre-osteoblast phenotype by affecting their anchoring in responding to zirconia.
The authors are grateful to Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP) for the financial support (grants: #2015/03639-8, 2016/08888-9, 2014/22689-3).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict ofinterest.
- 12.LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem U S. 1951;193:265–75.Google Scholar
- 14.Lefebvre V, Peeters-Joris C, Vaes G. Production of gelatin-degrading matrix metalloproteinases (’type IV collagenases’) and inhibitors by articular chondrocytes during their dedifferentiation by serial subcultures and under stimulation by interleukin-1 and tumor necrosis factor alpha. Biochim Biophys Acta Neth. 1991;1094:8–18.CrossRefGoogle Scholar
- 18.Cavagis A, Takamori E, Granjeiro J, Oliveira R, Ferreira C, Peppelenbosch M, et al. TNFalpha contributes for attenuating both Y397FAK and Y416Src phosphorylations in osteoblasts. Oral Dis Den. 2014;20:780–6.Google Scholar
- 21.Dixon RDS, Chen Y, Ding F, Khare SD, Prutzman KC, Schaller MD et al. New Insights into FAK signaling and localization based on detection of a FAT domain folding intermediate. Chapel Hill: University of North Carolina; 2004;12:2161–71.Google Scholar
- 25.Fang X, Liu X, Yao L, Chen C, Lin J, Ni P, et al. New insights into FAK phosphorylation based on a FAT domain-defective mutation. PloS One. 2014;9:1–10.Google Scholar
- 38.Panagakos FS, Kumar S. Differentiation of human osteoblastic cells in culture: modulation of proteases by extracellular matrix and tumor necrosis factor-alpha. Inflammation. U S. 1995;19:423–43.Google Scholar
- 39.Wang R, Wang W, Ao L, Wang Z, Hao X, Zhang H. Benzo[a]pyrene-7,8-diol-9,10-epoxide suppresses the migration and invasion of human extravillous trophoblast HTR-8/SVneo cells by down-regulating MMP2 through inhibition of FAK/SRC/PI3K/AKT pathway. Toxicol Elsevier. 2017;386:72–83.CrossRefGoogle Scholar
- 41.Oum’hamed Z, Garnotel R, Josset Y, Trenteseaux C, Laurent-Maquin D. Matrix metalloproteinases MMP-2, -9 and tissue inhibitors TIMP-1, -2 expression and secretion by primary human osteoblast cells in response to titanium, zirconia, and alumina ceramics. J Biomed Mater Res. 2004;68:114–22.CrossRefGoogle Scholar