Differential gene expression response of synovial fibroblasts from temporomandibular joints and knee joints to dynamic tensile stress

Nazet, Ute; Neubert, Patrick; Schatz, Valentin; Grässel, Susanne; Proff, Peter; Schröder, Agnes; Kirschneck, Christian

doi:10.1007/s00056-021-00309-y

Cited by 4 publications

(9 citation statements)

References 55 publications

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“…The mice were euthanised and dissected conforming to national and institutional regulations. The cells have been isolated and were characterized previously [ 41 ]. The synovial fibroblasts were cultured at 37 °C in RPMI1640 with GlutaMAX™ (61870-010, Gibco, Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal calf serum (FCS, P30-3302, PAN-Biotech, Aidenbach, Germany), 1% antifungal/antibiotic (A5955, Sigma-Aldrich, Darmstadt, Germany), and 1% ascorbic acid (A8960, Sigma-Aldrich).…”

Section: Methodsmentioning

confidence: 99%

“…The qPCR protocol basically followed the previously published proceedings [ 41 , 44 ]. Briefly, RNA was extracted using RNA-Solv (R6830-01, VWR, Darmstadt, Germany) according to the manufacturer’s protocol.…”

Section: Methodsmentioning

confidence: 99%

“…A total of 200,000 primary synovial knee fibroblasts were seeded per well on collagencoated 6-well Bioflex plates (BF-3001C, Dunn Labortechnik, Asbach, Germany) and incubated overnight at 37 • C. On the following day, the culture medium was replaced by a medium with different sodium chloride concentrations (−20 mM, ±0 mM, +50 mM). Then, synovial fibroblasts were subjected to intermittent tensile strain (amplitude 15%; frequency 0.5 Hz; two eight-hour rest periods followed by 16 h of stretching; Figure 1b) [41,44,45]. The synovial fibroblasts were exposed to different sodium chloride concentrations for a total of 48 h.…”

Section: Experiments With Intermittent Tensile Strainmentioning

confidence: 99%

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Mechanical Stress Induces Sodium Entry and Osmoprotective Responses in Murine Synovial Fibroblasts

Proff,

Nazet,

Schröder

et al. 2024

Cells

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Osteoarthritis (OA) is a multifactorial disease depending on molecular, genetic, and environmental factors like mechanical strain. Next to the cartilage and the subchondral bone, OA also affects the synovium, which is critically involved in the maintenance of joint homeostasis. As there is a correlation between the extracellular sodium content in the knee joint and OA, this study investigates the impact of sodium on OA-associated processes like inflammation and bone remodeling without and with mechanical loading in synovial fibroblasts. For that purpose, murine synovial fibroblasts from the knee joint were exposed to three different extracellular sodium chloride concentrations (−20 mM, ±0 mM and +50 mM NaCl) in the absence or presence of compressive or intermittent tensile strain. In addition to the intracellular Na+ content and gene expression of the osmoprotective transcription factor nuclear factor of activated T cells 5 (Nfat5), the gene and protein expression of inflammatory mediators (interleukin-6 (IL6), prostaglandin endoperoxide synthase-2 (Ptgs2)/prostaglandin E2 (PGE2)), and factors involved in bone metabolism (receptor activator of NF-κB ligand (RANKL), osteoprotegerin (OPG)) were analyzed by qPCR and ELISA. Mechanical strain already increased intracellular Na+ and Nfat5 gene expression at standard salt conditions to levels obtained by exposure to increased extracellular Na+ content. Both high salt and compressive strain resulted in elevated IL6 and PGE2 release. Intermittent tensile strain did not increase Il6 mRNA expression or IL6 protein secretion but triggered Ptgs2 expression and PGE2 production. Increased extracellular Na+ levels and compressive strain increased RANKL expression. In contrast, intermittent tension suppressed RANKL expression without this response being subject to modification by extracellular sodium availability. OPG expression was only induced by compressive strain. Changes in extracellular Na+ levels modified the inflammatory response and altered the expression of mediators involved in bone metabolism in cells exposed to mechanical strain. These findings indicate that Na+ balance and Nfat5 are important players in synovial fibroblast responses to mechanical stress. The integration of Na+ and Na+-dependent signaling will help to improve the understanding of the pathogenesis of osteoarthritis and could lead to the establishment of new therapeutic targets.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Experiments With Intermittent Tensile Strainmentioning

confidence: 99%

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Mechanical Stress Induces Sodium Entry and Osmoprotective Responses in Murine Synovial Fibroblasts

Proff,

Nazet,

Schröder

et al. 2024

Cells

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“…Additionally, a racehorse model was established as a specific joint-loading model used to measure the impact of effort on knee OA development, since microstructural changes in articular cartilage due to overloading of the equine joint were evidenced [ 44 , 45 ]. As the OA may progress differently in different joints [ 46 ], more research is still needed in equine TMJ OA, especially in the area of markers and signaling pathways.…”

Section: Tmj Oa In Humans and Horsesmentioning

confidence: 99%

Equine Models of Temporomandibular Joint Osteoarthritis: A Review of Feasibility, Biomarkers, and Molecular Signaling

Jasiński,

Turek,

Kaczorowski

et al. 2024

Biomedicines

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Osteoarthritis (OA) of the temporomandibular joint (TMJ) occurs spontaneously in humans and various animal species, including horses. In humans, obtaining tissue samples is challenging and clinical symptoms appear late in the disease progression. Therefore, genetically modified, induced, and naturally occurring animal models play a crucial role in understanding the pathogenesis and evaluating potential therapeutic interventions for TMJ OA. Among the naturally occurring models, the equine TMJ OA model is characterized by slow, age-related progression, a wide range of clinical examinations, and imaging modalities that can be performed on horses, as well as easy tissue and synovial fluid collection. The morphological and functional similarities of TMJ structures in both species make the equine model of TMJ OA an excellent opportunity to track disease progression and response to treatment. However, much work remains to be carried out to determine the utility of human TMJ OA biomarkers in horses. Among the main TMJ OA biomarkers, IL-1, IL-6, TGF-β, TNF-α, and PGE2 have been recently investigated in the equine model. However, the majority of biomarkers for cartilage degradation, chondrocyte hypertrophy, angiogenesis, and TMJ overload—as well as any of the main signaling pathways—have not been studied so far. Therefore, it would be advisable to focus further research on equine specimens, considering both mediators and signaling.

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“…However, a major gap remains in our understanding of the mechanobiology of OA synovium. Importantly, tensile strain loading of synovial cells in vitro has been shown to induce changes in extracellular matrix, inflammatory, and oxidative stress gene expression 19 – 22 , all of which could impact joint health and homeostasis. Therefore, our objective was to assess the physiologic responses of intact human synovial tissues to mechanical loading with cyclic tensile strain.…”

Section: Introductionmentioning

confidence: 99%

Tensile strain and altered synovial tissue metabolism in human knee osteoarthritis

Philpott

Birmingham

Fiset

et al. 2022

Sci Rep

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Synovium is critical for maintaining joint homeostasis and may contribute to mechanobiological responses during joint movement. We investigated mechanobiological responses of whole synovium from patients with late-stage knee osteoarthritis (OA). Synovium samples were collected during total knee arthroplasty and assigned to histopathology or cyclic 10% tensile strain loading, including (1) static (control); (2) low-frequency (0.3 Hz); and iii) high-frequency (1.0 Hz) for 30-min. After 6-h incubation, tissues were bisected for RNA isolation and immunostaining (3-nitrotyrosine; 3-NT). RNA sequencing was analyzed for differentially expressed genes and pathway enrichment. Cytokines and lactate were measured in conditioned media. Compared to controls, low-frequency strain induced enrichment of pathways related to interferon response, Fc-receptor signaling, and cell metabolism. High-frequency strain induced enrichment of pathways related to NOD-like receptor signaling, high metabolic demand, and redox signaling/stress. Metabolic and redox cell stress was confirmed by increased release of lactate into conditioned media and increased 3-NT formation in the synovial lining. Late-stage OA synovial tissue responses to tensile strain include frequency-dependent increases in inflammatory signaling, metabolism, and redox biology. Based on these findings, we speculate that some synovial mechanobiological responses to strain may be beneficial, but OA likely disturbs synovial homeostasis leading to aberrant responses to mechanical stimuli, which requires further validation.

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Differential gene expression response of synovial fibroblasts from temporomandibular joints and knee joints to dynamic tensile stress

Cited by 4 publications

References 55 publications

Mechanical Stress Induces Sodium Entry and Osmoprotective Responses in Murine Synovial Fibroblasts

Mechanical Stress Induces Sodium Entry and Osmoprotective Responses in Murine Synovial Fibroblasts

Equine Models of Temporomandibular Joint Osteoarthritis: A Review of Feasibility, Biomarkers, and Molecular Signaling

Tensile strain and altered synovial tissue metabolism in human knee osteoarthritis

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