Mechanical stress influences the viability and morphology of human parametrial ligament fibroblasts

Hu, Ming; Li, Hong; Hong, Shasha; Min, Jie; Zhao, Yang; Yang, Qing; Zhang, Qifan; Tang, Jianming; Li, Yang

doi:10.3892/mmr.2016.6052

Cited by 17 publications

(16 citation statements)

References 16 publications

(16 reference statements)

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“…MTT assays also did not yield any significant effect of etoricoxib on cell viability, whereas the compressive force applied significantly reduced cell viability, that is, mitochondrial enzymatic activity, as assessed by the MTT assay. We thus hypothesize that either the mechanical stress itself [73] or an accompanying lack of oxygen [74] in our in vitro model, which is also supposed to occur during orthodontic tooth movement in vivo [75], most likely attenuated PDL fibroblast cell metabolism to a significant degree, whereas cell death was not significantly induced.…”

Section: Discussionmentioning

confidence: 99%

Effects of the Highly COX-2-Selective Analgesic NSAID Etoricoxib on Human Periodontal Ligament Fibroblasts during Compressive Orthodontic Mechanical Strain

Kirschneck

Küchler

Wolf

et al. 2019

Mediators of Inflammation

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Human periodontal ligament (hPDL) fibroblasts play a major role during periodontitis and orthodontic tooth movement, mediating periodontal inflammation, osteoclastogenesis, and collagen synthesis. The highly COX-2-selective NSAID etoricoxib has a favorable systemic side effect profile and high analgesic efficacy, particularly for orthodontic pain. In this in vitro study, we investigated possible side effects of two clinically relevant etoricoxib concentrations on the expression pattern of mechanically strained hPDL fibroblasts and associated osteoclastogenesis in a model of simulated orthodontic compressive strain occurring during orthodontic tooth movement. hPDL fibroblasts were incubated for 72 h under physiological conditions with etoricoxib at 0 μM, 3.29 μM, and 5.49 μM, corresponding to clinically normal and subtoxic dosages, with and without mechanical strain by compression (2 g/cm2) for the final 48 h, simulating conditions during orthodontic tooth movement in compressive areas of the periodontal ligament. We then determined gene and/or protein expression of COX-2, IL-6, PG-E2, RANK-L, OPG, ALPL, VEGF-A, P4HA1, COL1A2, and FN1 via RT-qPCR, ELISA, and Western blot analyses as well as apoptosis, necrosis, cell viability, and cytotoxicity via FACS, MTT, and LDH assays. In addition, hPDL fibroblast-mediated osteoclastogenesis was assessed by TRAP staining in coculture with RAW267.4 cells for another 72 h. Gene and protein expression of all evaluated factors was significantly induced by the mechanical compressive strain applied. Etoricoxib at 3.29 μM and 5.49 μM significantly inhibited PG-E2 synthesis, but not COX-2 and IL-6 gene expression nor RANK-L-/OPG-mediated osteoclastogenesis or angiogenesis (VEGF-A). Extracellular matrix remodeling (COL1A2, FN1) and bone anabolism (ALPL), by contrast, were significantly stimulated particularly at 5.49 μM. In general, no adverse etoricoxib effects on hPDL fibroblasts regarding apoptosis, necrosis, cell viability, or cytotoxicity were detected. Clinically dosed etoricoxib, that is, a highly selective COX-2 inhibition, did not have substantial effects on hPDL fibroblast-mediated periodontal inflammation, extracellular matrix remodeling, RANK-L/OPG expression, and osteoclastogenesis during simulated orthodontic compressive strain.

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Section: Discussionmentioning

confidence: 99%

Effects of the Highly COX-2-Selective Analgesic NSAID Etoricoxib on Human Periodontal Ligament Fibroblasts during Compressive Orthodontic Mechanical Strain

Kirschneck

Küchler

Wolf

et al. 2019

Mediators of Inflammation

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“…Our group used the four‐point bending mechanics loading system to treat pelvic floor fibroblasts in the earlier studies. It was confirmed that the mechanical loading of suitable parameters could well simulate fibroblast stretch injury and cause cell damage 27–29 . Considering these research, we established a DRGs injury model in vitro that was stretched by a four‐point bending system in order to simulate pudendal nerve injury in the process of pregnancy and childbirth.…”

Section: Discussionmentioning

confidence: 92%

Protective effect and potential mechanism of Schwann cell‐derived exosomes on mechanical damage of rat dorsal root ganglion cells

Huang

et al. 2021

J of Obstet and Gynaecol

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Background Pudendal nerve (PN) injury was one of the most important pathogenesis of stress urinary incontinence (SUI). Schwann cell (SC)‐derived exosomes could promote axonal regeneration. Wnt protein could significantly promote axonal regeneration and participate in the regulation of proliferation and differentiation of neural stem cells. Therefore, we sought to determine whether SCs‐derived exosomes might also protect against damaged dorsal root ganglion cells (DRGs) through the Wnt/β‐catenin pathway. Material and Methods The DRGs injury model was fabricated using a four‐point bending system. The exosomes were separated from the SCs supernatant. XAV939, which was a small molecule inhibitor, was used to inhibit the Wnt/β‐catenin pathway. Next, Cell Counting Kit‐8 (CCK8) kit was used to detect cell activity. We evaluated the proliferative activity of DRG cells using the cell cycle and apoptosis detection kit. We assessed the cell apoptotic rates through the Annexin V/PE double staining. Finally, we detect the expression of downstream proteins of Wnt/β‐catenin pathway in DRG cells using western blotting. Results SC‐derived exosomes had protective effects on DRGs after mechanical damage, which could promote cell proliferation, transition of the cell cycle to the G2 phase, and inhibit cell apoptosis. Exogenous administration of XAV939 suppressed the promoting effect of SCs ‐derived exosomes on DRG cells and the expression of downstream proteins of Wnt/β‐catenin pathway in DRG cells was also suppressed. Conclusion These results suggested that SC‐derived exosomes have a repairing effect on DRG cells injury caused by cyclic mechanical stretching (CMS) and the Wnt/β‐catenin pathway is potentially involved in the process.

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“…The previous study revealed that mechanical forces significantly effect cell activity, such as reduced cell proliferation, mitochondria vacuolization, or the cell cytoskeleton depolymerization. Among these mechanical forces, shear stress and hydrostatic pressure directly affect the physical and biological characteristics of cells and become important participants in cell viability [40,41]. In the final part of the simulation, we mimicked conditions such as hydrostatic pressure, capillary phenomenon, and liquid disturbance during spheroid formation.…”

Section: Discussionmentioning

confidence: 99%

A Dynamic Hanging-Drop System for Mesenchymal Stem Cell Culture

Huang

Tzeng

Chen

et al. 2020

IJMS

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There have been many microfluid technologies combined with hanging-drop for cell culture gotten developed in the past decade. A common problem within these devices is that the cell suspension introduced at the central inlet could cause a number of cells in each microwell to not regularize. Also, the instability of droplets during the spheroid formation remains an unsolved ordeal. In this study, we designed a microfluidic-based hanging-drop culture system with the design of taper-tube that can increase the stability of droplets while enhancing the rate of liquid exchange. A ring is surrounding the taper-tube. The ring can hold the cells to enable us to seed an adequate amount of cells before perfusion. Moreover, during the period of cell culture, the mechanical force around the cell is relatively low to prevent stem cells from differentiate and maintain the phenotype. As a result of our hanging system design, cells are designed to accumulate at the bottom of the droplet. This method enhances convenience for observation activities and analysis of experiments. Thus, this microfluid chip can be used as an in vitro platform representing in vivo physiological conditions, and can be useful in regenerative therapy.

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Mechanical stress influences the viability and morphology of human parametrial ligament fibroblasts

Cited by 17 publications

References 16 publications

Effects of the Highly COX-2-Selective Analgesic NSAID Etoricoxib on Human Periodontal Ligament Fibroblasts during Compressive Orthodontic Mechanical Strain

Effects of the Highly COX-2-Selective Analgesic NSAID Etoricoxib on Human Periodontal Ligament Fibroblasts during Compressive Orthodontic Mechanical Strain

Protective effect and potential mechanism of Schwann cell‐derived exosomes on mechanical damage of rat dorsal root ganglion cells

A Dynamic Hanging-Drop System for Mesenchymal Stem Cell Culture

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