The aim of this study was to investigate the response of human periodontal ligament (hPDL) fibroblasts to an intermittent compressive force and its effect on the expression of SOST, POSTN, and TGFB1. A computerized cell compressive force loading apparatus was introduced, and hPDL cells were subjected to intermittent compressive force. The changes in messenger RNA (mRNA) and protein expression were monitored by real-time polymerase chain reaction and Western blot analysis, respectively. An increased expression of SOST, POSTN, and TGFB1 was observed in a time-dependent fashion. Addition of cycloheximide, a transforming growth factor (TGF)-β inhibitor (SB431542), or a neutralizing antibody against TGF-β1 attenuated the force-induced expression of SOST and POSTN as well as sclerostin and periostin, indicating a role of TGF-β1 in the pressure-induced expression of these proteins. Enzyme-linked immunosorbent assay analysis revealed an increased level of TGF-β1 in the cell extracts but not in the medium, suggesting that intermittent compressive force promoted the accumulation of TGF-β1 in the cells or their surrounding matrix. In conclusion, an intermittent compressive force regulates SOST/POSTN expression by hPDL cells via the TGF-β1 signaling pathway. Since these proteins play important roles in the homeostasis of the periodontal tissue, our results indicate the importance of masticatory forces in this process.
Mechanical force regulates periodontal ligament cell (PDL) behavior. However, different force types lead to distinct PDL responses. Here, we report that pretreatment with an intermittent compressive force (ICF), but not a continuous compressive force (CCF), promoted human PDL (hPDL) osteogenic differentiation as determined by osteogenic marker gene expression and mineral deposition in vitro. ICF-induced osterix (OSX) expression was inhibited by cycloheximide and monensin. Although CCF and ICF significantly increased extracellular adenosine triphosphate (ATP) levels, pretreatment with exogenous ATP did not affect hPDL osteogenic differentiation. Gene-expression profiling of hPDLs subjected to CCF or ICF revealed that extracellular matrix (ECM)-receptor interaction, focal adhesion, and transforming growth factor beta (TGF-β) signaling pathway genes were commonly upregulated, while calcium signaling pathway genes were downregulated in both CCF-and ICF-treated hPDLs. The TGFB1 mRNA level was significantly increased, while those of TGFB2 and TGFB3 were decreased by ICF treatment. In contrast, CCF did not modify TGFB1 expression. Inhibiting TGF-β receptor type I or adding a TGF-β1 neutralizing antibody attenuated the ICF-induced OSX expression. Exogenous TGF-β1 pretreatment promoted hPDL osteogenic marker gene expression and mineral deposition. Additionally, pretreatment with ICF in the presence of TGF-β receptor type I inhibitor attenuated the ICF-induced mineralization. In conclusion, this study reveals the effects of ICF on osteogenic differentiation in hPDLs and implicates TGF-β signaling as one of its regulatory mechanisms.
The data contained within this article relate to a rotating rod within a stationary ring that was used to generate shear stress on cells and tissues via a medium. The geometry of the rotating rod within a stationary ring was designed to work with a 35-mm diameter culture dish. The data of the shear stress distribution are presented in terms of area-weighted average shear stress and the uniformity index, which were calculated for medium volumes of 4 and 5 ml at various rotational speeds ranging from 0 to 1000 rpm.
Objectives: Mechanical injury of dental pulp leads to root resorption by osteoclasts/ odontoclasts. S100 proteins have been demonstrated to be involved in inflammatory processes and bone remodeling. This study aimed to investigate the effect of mechanical stress on S100A7 expression by human dental pulp cells (HDPCs) and the effect of S100A7 proteins on osteoclast differentiation.
Materials and Methods: Isolated HDPCs were stimulated with compressive loading(2 and 6 hr), or shear loading (2, 6, and 16 hr). S100 mRNA expression and S100A7 protein levels were determined by real-time PCR and ELISA, respectively. Osteoclast differentiation was analyzed using primary human monocytes. The differentiation and activity of osteoclasts were examined by TRAcP staining and dentine resorption.In addition, the expression of S100A7 was analyzed in pulp tissues obtained from orthodontically treated teeth.
Results:Compressive and shear mechanical stress significantly upregulated both mRNA and protein level of S100A7. Dental pulp tissues from orthodontically treated teeth exhibited higher S100A7 mRNA levels compared to non-treated control teeth. S100A7 promoted osteoclast differentiation by primary human monocytes.Moreover, S100A7 significantly enhanced dentine resorption by these cells.
Conclusions:Mechanical stress induced expression of S100A7 by human dental pulp cells and this may promote root resorption by inducing osteoclast differentiation and activity.
K E Y W O R D Sdental pulp, inflammation, mechanical stress, Osteoclast, S100 protein, S100A7 | 813 CHAROENPONG Et Al.
A polymer electrolyte fuel cell membrane electrode assembly (PEFC MEA) model that focuses on the structural and reaction parameters of catalyst layers has been developed. The reaction and structural parameters were modeled independently by considering the oxygen reduction reaction (ORR) activity in terms of current per unit active surface area of Pt [A cm -2 -Pt]. The catalyst layer models were constructed based on an assumption of cylindrical secondary pore structure, which was verified by measuring the primary pore size using mercury porosimetry. We found that penetration of Nafion ® electrolyte into the primary pores of Pt/C catalysts was restricted, and thus diffusion and reaction in primary pores became negligible. Moreover, the experimental results demonstrated that for the same catalyst, ORR activity remained almost constant, irrespective of the agglomerate size (i.e., 2.7 × × × × × 10 -6 A cm -2 -Pt at 60°C for Pt/C TEC10E50E in this study). This supported our idea that reaction parameters and structural parameters should be considered independently in PEFC modeling. The cell performance predicted with the developed model was satisfactorily accurate as compared to that obtained from experiments. As a result, the modeling developed in this study can be used to construct simple PEFC models that yield results with good accuracy and can be a useful tool for the development of PEFCs in future.
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