Therapeutic ultrasound stimulates MC3T3-E1 cell proliferation through the activation of NF-κB1, p38α, and mTOR

Tassinary, João Alberto Fioravante; Lunardelli, Adroaldo; Basso, Bruno Souza; Stülp, Simone; Pozzobon, Adriane; Pedrazza, Leonardo; Bartrons, Ramón; Ventura, Francesc; Rosa, José Luís; Melo, Denizar Alberto da Silva; Nunes, Fernanda Bordignon; Donadio, Márcio Vinı́cius Fagundes; Oliveira, Jarbas Rodrigues de

doi:10.1002/lsm.22414

Cited by 11 publications

(11 citation statements)

References 23 publications

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“…A previous study also suggested that LIPUS did not have any changes in temperature. The main effect of LIPUS could be caused by “nonthermal effects,” such as cavitation effects and microstreaming effects …”

Section: Discussionmentioning

confidence: 99%

LIPUS inhibited the expression of inflammatory factors and promoted the osteogenic differentiation capacity of hPDLCs by inhibiting the NF‐κB signaling pathway

Liu

Zhou

et al. 2019

J of Periodontal Research

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Background and Objectives As a chronic infectious disease, periodontitis could lead to tooth and bone loss. Low‐intensity pulsed ultrasound (LIPUS) is a safe, noninvasive treatment method to effectively inhibit inflammation and promote bone differentiation. However, the application of LIPUS in curing periodontitis is still rare. Our study aimed to explore the ability of LIPUS to inhibit inflammatory factors and promote the osteogenic differentiation capacity of human periodontal ligament cells (hPDLCs), and its underlying mechanism. Material and Methods Human periodontal ligament cells were obtained and cultured from the premolar tissue samples for experiments. First, hPDLCs were treated for 24 hours using lipopolysaccharide (LPS) and then exposed to LIPUS (10 mW/cm2, 30 mW/cm2, 60 mW/cm2, and 90 mW/cm2) to determine the appropriate intensity to inhibit expression of the inflammatory factors interleukin‐6 (IL‐6) and interleukin‐8 (IL‐8) expression. The expression of IL‐6 and IL‐8 was detected by real‐time PCR and enzyme‐linked immunosorbent assay. The safety of the most appropriate intensity of LIPUS was tested by a cell counting kit 8 test and an apoptosis assay. Then, LPS‐induced hPDLCs were treated in osteogenic medium for 7‐21 days with or without LIPUS (90 mW/cm2, 30 min/d) stimulation. The osteogenic genes RUNX2, OPN, OSX, and OCN were measured by real‐time PCR. Additionally, osteogenic differentiation capacity was determined using alkaline phosphatase (ALP) staining, ALP activity analysis, and Alizarin red staining. The activity of the nuclear factor‐kappa B (NF‐κB) signaling pathway was determined by western blotting, real‐time PCR, immunofluorescence, and pathway blockade assays. Results Lipopolysaccharide significantly upregulated the production and gene expression of IL‐6 and IL‐8, while LIPUS stimulation significantly inhibited IL‐6 and IL‐8 expression in an intensity‐dependent manner. LIPUS (90 mW/cm2) was chosen as the most appropriate intensity, and there was no detrimental influence on cell proliferation and status with or without osteogenic medium. In addition, consecutive stimulation with LIPUS (90 mW/cm2) for 30 min/d for 7 days could also inhibit IL‐6 and IL‐8 gene expression, upregulate the expression of the osteogenesis‐related genes RUNX2, OPN, OSX, and OCN, and promote osteogenic differentiation capacity in osteogenic medium in inflamed hPDLCs. The NF‐κB signaling pathway was inhibited with LIPUS (90 mW/cm2) via inhibition of the phosphorylation of IκBα and the translocation of p65 into the nucleus in inflamed hPDLCs. Additional investigations of the NF‐κB inhibitor, BAY 11‐7082, revealed that LIPUS (90 mW/cm2) acted similarly to BAY 11‐7802 to inhibit the NF‐κB signaling pathway and increase osteogenesis‐related genes and promote the osteogenic differentiation capacity of inflamed hPDLCs. Conclusion Low‐intensity pulsed ultrasound (90 mW/cm2) stimulation could be a safe method to inhibit IL‐6 and IL‐8 in hPDLCs by inhibiting the NF‐κB signaling pathway. The effect of LIPUS (90 mW...

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Section: Discussionmentioning

confidence: 99%

LIPUS inhibited the expression of inflammatory factors and promoted the osteogenic differentiation capacity of hPDLCs by inhibiting the NF‐κB signaling pathway

Liu

Zhou

et al. 2019

J of Periodontal Research

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“…ERK1/2 pathway involvement was previously observed 8,9 . The p38 MAPK pathway, which is another significant subgroup of the MAPK superfamily, was involved in the response to proliferation-inducing stimuli in an osteoblastic cell line 11 . In addition, according to Sinfield's study, the p38α subtype facilitates IL-1-induced IL-6 secretion by human cardiac fibroblasts 12 .…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, we investigated IκBα and pp65 as key factors to indirectly assess NF-κB activation. Notably, MAPKs, including ERK and p38, are typically involved in bone metabolism in combination with NF-κB pathway activation 11,19 . Both pathways are associated with the inhibition of osteoclast formation by K7 in mouse bone marrow-derived macrophages 19 as well as the therapeutic ultrasound-induced proliferation of MC3T3-E1 cells 11 .…”

Section: Discussionmentioning

confidence: 99%

“…The p38 pathway is a stress response signaling pathway that is activated by intracellular and extracellular stresses, including environmental changes and exposure to inflammatory cytokines or other biosynthetic stresses 10 . The p38 pathway is thus involved in variety of biological responses as well as molecular regulation 11,12 . We hypothesized a role for p38 in IL-6 secretion by MLO-Y4 cells stimulated by CCF.…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, NF-κB is a nuclear transcription factor that is activated rapidly by different stimuli, such as inflammatory cytokines, including IL-1 and TNF-α; a number of growth factors; and mechanical stress inducers 15,16 . Furthermore, both p38 and NF-κB exert synergistic regulatory and biological effects on bone metabolism 11,[17][18][19] . The purpose of this study was to therefore investigate whether p38 and NF-κB are activated during CCF-stimulated IL-6 secretion in MLO-Y4 cells.…”

Section: Introductionmentioning

confidence: 99%

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The p38 MAPK and NF-κB Pathways are Involved in Cyclic Compressive Force-induced IL-6 Secretion in MLO-Y4 Cells

Chen

et al. 2018

Braz. arch. biol. technol.

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We previously revealed the involvement of extracellular regulated protein kinases 1/2 (ERK1/2) in interleukin-6 (IL-6) secretion induced by cyclic compressive force (CCF) in MLO-Y4 cells. In this study, we investigated the contributions of the p38 mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) pathways to IL-6 secretion by stimulating MLO-Y4 cells with CCF. At 80% confluence, different magnitudes (1000μstrain, 2000 μstrain and 4000 μstrain), frequencies (0.5 Hz, 1.0 Hz and 2.0 Hz) and durations (10 min, 30 min, 1 h, 3 h and 6 h) of CCF were loaded onto cells using a four-point bending system. Flow Cytometry (FCM) analysis was used to analyze cell mortality rates after CCF loading. p38 and p65 phosphorylation as well as IκBα degradation in MLO-Y4 cells were detected by Western blotting (WB). Changes in IL-6 secretion after inhibitor treatment were assessed by enzyme-linked immunosorbent assays (ELISAs). Cellular viability was over 90 percent after CCF. p38 and p65 phosphorylation increased under all conditions, whereas IκBα protein levels decreased. However, phosphorylation and degradation were not completely dependent on the loading magnitude, frequency or duration. Furthermore, p38 inhibition using the specific inhibitor SB203580 reduced both p38 phosphorylation and IL-6 secretion. Similarly, NF-κB inhibition using BAY 11-7082 decreased p65 phosphorylation and IL-6 secretion but increased the concentration of IκBα. These findings reveal significant roles for the p38 and NF-κB signaling pathways in IL-6 secretion induced by CCF in MLO-Y4 cells.

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Mammalian target of rapamycin as a therapeutic target in osteoporosis

Shen

Ren²,

Qiu

et al. 2017

Journal Cellular Physiology

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The mechanistic target of rapamycin (mTOR) plays a key role in sensing and integrating large amounts of environmental cues to regulate organismal growth, homeostasis, and many major cellular processes. Recently, mounting evidences highlight its roles in regulating bone homeostasis, which sheds light on the pathogenesis of osteoporosis. The activation/inhibition of mTOR signaling is reported to positively/negatively regulate bone marrow mesenchymal stem cells (BMSCs)/osteoblasts-mediated bone formation, adipogenic differentiation, osteocytes homeostasis, and osteoclasts-mediated bone resorption, which result in the changes of bone homeostasis, thereby resulting in or protect against osteoporosis. Given the likely importance of mTOR signaling in the pathogenesis of osteoporosis, here we discuss the detailed mechanisms in mTOR machinery and its association with osteoporosis therapy.

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Therapeutic ultrasound stimulates MC3T3-E1 cell proliferation through the activation of NF-κB1, p38α, and mTOR

Cited by 11 publications

References 23 publications

LIPUS inhibited the expression of inflammatory factors and promoted the osteogenic differentiation capacity of hPDLCs by inhibiting the NF‐κB signaling pathway

LIPUS inhibited the expression of inflammatory factors and promoted the osteogenic differentiation capacity of hPDLCs by inhibiting the NF‐κB signaling pathway

The p38 MAPK and NF-κB Pathways are Involved in Cyclic Compressive Force-induced IL-6 Secretion in MLO-Y4 Cells

Mammalian target of rapamycin as a therapeutic target in osteoporosis

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