scholarly journals Mechanical Loading of Cartilage Explants with Compression and Sliding Motion Modulates Gene Expression of Lubricin and Catabolic Enzymes

Cartilage ◽  
2015 ◽  
Vol 6 (3) ◽  
pp. 185-193 ◽  
Author(s):  
Oliver R. Schätti ◽  
Michala Marková ◽  
Peter A. Torzilli ◽  
Luigi M. Gallo
Keyword(s):  
Gene Expression ◽  
Mechanical Loading ◽  
Cartilage Explants ◽  
Catabolic Enzymes ◽  
Sliding Motion

scholarly journals IL-1β Damages Fibrocartilage and Upregulates MMP-13 Expression in Fibrochondrocytes in the Condyle of the Temporomandibular Joint

2019 ◽  
Vol 20 (9) ◽  
pp. 2260 ◽  
Author(s):  
Hessam Tabeian ◽  
Beatriz F. Betti ◽  
Cinthya dos Santos Cirqueira ◽  
Teun J. de Vries ◽  
Frank Lobbezoo ◽  
...  
Keyword(s):  
Gene Expression ◽  
Temporomandibular Joint ◽  
Mechanical Loading ◽  
Cartilage Damage ◽  
Cartilage Degradation ◽  
Mechanical Stimuli ◽  
Condylar Cartilage ◽  
Catabolic Enzymes ◽  
Mechanical Tensile ◽  
And Function

The temporomandibular joint (TMJ), which differs anatomically and biochemically from hyaline cartilage-covered joints, is an under-recognized joint in arthritic disease, even though TMJ damage can have deleterious effects on physical appearance, pain and function. Here, we analyzed the effect of IL-1β, a cytokine highly expressed in arthritic joints, on TMJ fibrocartilage-derived cells, and we investigated the modulatory effect of mechanical loading on IL-1β-induced expression of catabolic enzymes. TMJ cartilage degradation was analyzed in 8–11-week-old mice deficient for IL-1 receptor antagonist (IL-1RA−/−) and wild-type controls. Cells were isolated from the juvenile porcine condyle, fossa, and disc, grown in agarose gels, and subjected to IL-1β (0.1–10 ng/mL) for 6 or 24 h. Expression of catabolic enzymes (ADAMTS and MMPs) was quantified by RT-qPCR and immunohistochemistry. Porcine condylar cells were stimulated with IL-1β for 12 h with IL-1β, followed by 8 h of 6% dynamic mechanical (tensile) strain, and gene expression of MMPs was quantified. Early signs of condylar cartilage damage were apparent in IL-1RA−/− mice. In porcine cells, IL-1β strongly increased expression of the aggrecanases ADAMTS4 and ADAMTS5 by fibrochondrocytes from the fossa (13-fold and 7-fold) and enhanced the number of MMP-13 protein-expressing condylar cells (8-fold). Mechanical loading significantly lowered (3-fold) IL-1β-induced MMP-13 gene expression by condylar fibrochondrocytes. IL-1β induces TMJ condylar cartilage damage, possibly by enhancing MMP-13 production. Mechanical loading reduces IL-1β-induced MMP-13 gene expression, suggesting that mechanical stimuli may prevent cartilage damage of the TMJ in arthritic patients.

Development of a Porcine Model to Assess the Effect of In-Situ Knee Joint Loading On Site-Specific Cartilage Gene Expression

2021 ◽  
Author(s):  
Baaba Otoo ◽  
LePing Li ◽  
David A. Hart ◽  
Walter Herzog
Keyword(s):  
Gene Expression ◽  
Mechanical Loading ◽  
Fluid Pressure ◽  
Cartilage Explants ◽  
Knee Joints ◽  
Expression Levels ◽  
Biological Responses ◽  
Loading Tests ◽  
Gene Expression Levels

Abstract Cyclic mechanical loading of cartilage induces stresses and fluid flow which are thought to modulate chondrocyte metabolism. The uneven surface, plus the heterogeneity of cartilage within a joint, makes stress and fluid pressure distribution in the tissue non-uniform, and gene expression may vary at different sites as a function of load magnitude, frequency and time. In previous studies, cartilage explants were used for loading tests to investigate biological responses of the cartilage to mechanical loading. In contrast, we used loading tests on intact knee joints, to better reflect the loading conditions in a joint, and thus provide a more physiologically relevant mechanical environment. Gene expression levels in loaded samples for a selection of relevant genes were compared with those of the corresponding unloaded control samples to characterize potential differences. Furthermore, the effect of load magnitude and duration on gene expression levels were investigated. We observed differences in gene expression levels between samples from different sites in the same joint and between corresponding samples from the same site in loaded and unloaded joints. Consistent with previous findings, our results indicate that there is a critical upper and lower threshold of loading for triggering the expression of certain genes. Variations in gene expression levels may reflect the effect of local loading, topography and structure of the cartilage in an intact joint on the metabolic activity of the associated cells.

Anti-Inflammatory Effects of Novel Standardized Platelet Rich Plasma Releasates on Knee Osteoarthritic Chondrocytes and Cartilage in vitro

2019 ◽  
Vol 20 (11) ◽  
pp. 920-933 ◽  
Author(s):  
Lucía Gato-Calvo ◽  
Tamara Hermida-Gómez ◽  
Cristina R. Romero ◽  
Elena F. Burguera ◽  
Francisco J. Blanco
Keyword(s):  
Gene Expression ◽  
Ex Vivo ◽  
Platelet Rich Plasma ◽  
Cartilage Explants ◽  
Ex Vivo Model ◽  
Chondrocyte Viability ◽  
Platelet Concentration ◽  
The Absolute ◽  
Anti Inflammatory

Background: Platelet Rich Plasma (PRP) has recently emerged as a potential treatment for osteoarthritis (OA), but composition heterogeneity hampers comparison among studies, with the result that definite conclusions on its efficacy have not been reached. Objective: 1) To develop a novel methodology to prepare a series of standardized PRP releasates (PRP-Rs) with known absolute platelet concentrations, and 2) To evaluate the influence of this standardization parameter on the anti-inflammatory properties of these PRP-Rs in an in vitro and an ex vivo model of OA. Methods: A series of PRPs was prepared using the absolute platelet concentration as the standardization parameter. Doses of platelets ranged from 0% (platelet poor plasma, PPP) to 1.5·105 platelets/µl. PRPs were then activated with CaCl2 to obtain releasates (PRP-R). Chondrocytes were stimulated with 10% of each PRP-R in serum-free culture medium for 72 h to assess proliferation and viability. Cells were co-stimulated with interleukin (IL)-1β (5 ng/ml) and 10% of each PRP-R for 48 h to determine the effects on gene expression, secretion and intra-cellular content of common markers associated with inflammation, catabolism and oxidative stress in OA. OA cartilage explants were co-stimulated with IL-1β (5 ng/ml) and 10% of either PRP-R with 0.75·105 platelets/µl or PRP-R with 1.5·105 platelets/µl for 21 days to assess matrix inflammatory degradation. Results: Chondrocyte viability was not affected, and proliferation was dose-dependently increased. The gene expression of all pro-inflammatory mediators was significantly and dose-independently reduced, except for that of IL-1β and IL-8. Immunoblotting corroborated this effect for inducible NO synthase (NOS2). Secreted matrix metalloproteinase-13 (MMP-13) was reduced to almost basal levels by the PRP-R from PPP. Increasing platelet dosage led to progressive loss to this anti-catabolic ability. Safranin O and toluidine blue stains supported the beneficial effect of low platelet dosage on cartilage matrix preservation. Conclusion: We have developed a methodology to prepare PRP releasates using the absolute platelet concentration as the standardization parameter. Using this approach, the composition of the resulting PRP derived product is independent of the donor initial basal platelet count, thereby allowing the evaluation of its effects objectively and reproducibly. In our OA models, PRP-Rs showed antiinflammatory, anti-oxidant and anti-catabolic properties. Platelet enrichment could favor chondrocyte proliferation but is not necessary for the above effects and could even be counter-productive.

Mechanical loading-induced gene expression and BMD changes are different in two inbred mouse strains

2005 ◽  
Vol 99 (5) ◽  
pp. 1951-1957 ◽  
Author(s):  
Chandrasekhar Kesavan ◽  
Subburaman Mohan ◽  
Susanna Oberholtzer ◽  
Jon E. Wergedal ◽  
David J. Baylink
Keyword(s):  
Gene Expression ◽  
Mechanical Loading ◽  
Quantitative Computed Tomography ◽  
Age Groups ◽  
Marker Genes ◽  
Type I ◽  
Mineral Density ◽  
Anabolic Response ◽  
Bone Response ◽  
C3h Mice

Our goal is to evaluate skeletal anabolic response to mechanical loading in different age groups of C57B1/6J (B6) and C3H/HeJ (C3H) mice with variable loads using bone size, bone mineral density (BMD), and gene expression changes as end points. Loads of 6–9 N were applied at 2 Hz for 36 cycles for 12 days on the tibia of 10-wk-old female B6 and C3H mice. Effects of a 9-N load on 10-, 16-, and 36-wk-old C3H mice were also studied. Changes in bone parameters were measured using peripheral quantitative computed tomography, and gene expression was determined by real-time PCR. Total volumetric BMD was increased by 5 and 15%, respectively, with 8- and 9-N loads in the B6, but not the C3H, mice. Increases of 20 and 12% in periosteal circumference were reflected by dramatic 44 and 26% increases in total area in B6 and C3H mice, respectively. The bone response to bending showed no difference in the three age groups of B6 and C3H mice. At 2 days, mechanical loading resulted in significant downregulation in expression of bone resorption (BR), but not bone formation (BF) marker genes. At 4 and 8 days of loading, expression of BF marker genes (type I collagen, alkaline phosphatase, osteocalcin, and bone sialoprotein) was increased two- to threefold and expression of BR marker genes (matrix metalloproteinase-9 and thrombin receptor-activating peptide) was decreased two- to fivefold. Although expression of BF marker genes was upregulated four- to eightfold at 12 days of training, expression of BR marker genes was upregulated seven- to ninefold. Four-point bending caused significantly greater changes in expression of BF and BR marker genes in bones of the B6 than the C3H mice. We conclude that mechanical loading-induced molecular pathways are activated to a greater extent in the B6 than in the C3H mice, resulting in a higher anabolic response in the B6 mice.

scholarly journals Mechanical Loading in Osteocytes Induces Formation of a Src/Pyk2/MBD2 Complex That Suppresses Anabolic Gene Expression

PLoS ONE ◽  
2014 ◽  
Vol 9 (5) ◽  
pp. e97942 ◽  
Author(s):  
Julia M. Hum ◽  
Richard N. Day ◽  
Joseph P. Bidwell ◽  
Yingxiao Wang ◽  
Fredrick M. Pavalko
Keyword(s):  
Gene Expression ◽  
Mechanical Loading

Influence of a single loading episode on gene expression in healing rat Achilles tendons

2012 ◽  
Vol 112 (2) ◽  
pp. 279-288 ◽  
Author(s):  
Pernilla Eliasson ◽  
Therese Andersson ◽  
Per Aspenberg
Keyword(s):  
Gene Expression ◽  
Mechanical Loading ◽  
Tendon Healing ◽  
Oxygen Species ◽  
Secondary Effects ◽  
Gene Expression Response ◽  
Affymetrix Microarray ◽  
Healing Tendon ◽  
Expression Response ◽  
Stimulation Of

Mechanical loading stimulates tendon healing via mechanisms that are largely unknown. Genes will be differently regulated in loaded healing tendons, compared with unloaded, just because of the fact that healing processes have been changed. To avoid such secondary effects and study the effect of loading per se, we therefore studied the gene expression response shortly after a single loading episode in otherwise unloaded healing tendons. The Achilles tendon was transected in 30 tail-suspended rats. The animals were let down from the suspension to load their tendons on a treadmill for 30 min once, 5 days after tendon transection. Gene expression was studied by Affymetrix microarray before and 3, 12, 24, and 48 h after loading. The strongest response in gene expression was seen 3 h after loading, when 150 genes were up- or downregulated (fold change ≥2, P ≤ 0.05). Twelve hours after loading, only three genes were upregulated, whereas 38 were downregulated. Fewer than seven genes were regulated after 24 and 48 h. Genes involved in the inflammatory response were strongly regulated at 3 and 12 h after loading; this included upregulation of iNOS, PGE synthase, and IL-1β. Also genes involved in wound healing/coagulation, angiogenesis, and production of reactive oxygen species were strongly regulated by loading. Microarray results were confirmed for 16 selected genes in a repeat experiment ( N = 30 rats) using real-time PCR. It was also confirmed that a single loading episode on day 5 increased the strength of the healing tendon on day 12. In conclusion, the fact that there were hardly any regulated genes 24 h after loading suggests that optimal stimulation of healing requires a mechanical loading stimulus every day.

scholarly journals Mechanical injury of cartilage explants causes specific time-dependent changes in chondrocyte gene expression

2005 ◽  
Vol 52 (8) ◽  
pp. 2386-2395 ◽  
Author(s):  
Jennifer H. Lee ◽  
Jonathan B. Fitzgerald ◽  
Michael A. DiMicco ◽  
Alan J. Grodzinsky
Keyword(s):  
Gene Expression ◽  
Mechanical Injury ◽  
Time Dependent ◽  
Cartilage Explants ◽  
Specific Time

scholarly journals Different Cyclic Mechanical Loading Patterns Alter Expression of Osteogenic Markers in a Fibroblast-Osteoblast Co-Culture Model Resembling the Tendon-To-Bone Interface After Autologous ACL Reconstruction

2014 ◽  
Vol 2 (11_suppl3) ◽  
pp. 2325967114S0019
Author(s):  
Philip Peter Roessler ◽  
Johannes Struewer ◽  
Jürgen Rudolph Paletta ◽  
Turgay Efe
Keyword(s):  
Gene Expression ◽  
Mechanical Loading ◽  
Mechanical Load ◽  
Cruciate Ligament ◽  
Culture Conditions ◽  
Bone Morphogenetic Protein 2 ◽  
Marker Genes ◽  
Bone Interface ◽  
Loading Patterns

Objectives: The aim of the study was to evaluate the influence of different cyclical mechanical loading patterns on co-cultures of fibroblasts and osteoblasts in vitro, simulating the conditions of the tendon-to-bone interface after anterior cruciate ligament reconstruction. Methods: Tendon-derived rodent fibroblasts (TDF) and osteoblast-like cells (OBL) were co-cultured to simulate the tendon-to-bone interface. Cyclical loading was applied for one hour twice a day for three days, with a frequency of 1 Hz and 3 % strain. Alkaline phosphatase (AP), osteocalcin (OC), collagen type 1 (COL1A1), and bone morphogenetic protein 2 (BMP-2) gene expression and protein deposition were detected by real-time polymerase chain reaction (qPCR) and immunocytochemical analysis. Results: Mechanical loading significantly decreased AP, OC, and COL1A1 gene expression in both OBL and TDF, compared to non-loaded cultures. However, mechanical load increased gene expression of the same marker genes including BMP-2 during co-culture. Immunocytochemistry demonstrated increased deposition of corresponding proteins in the same range, independent of culture conditions. Higher depositions of BMP-2 were shown under loading conditions for osteoblast and TDF monocultures. Prolongation of mechanical loading resulted in cell detachment and spheroid formation. Conclusion: Cyclical mechanical loading caused downregulation of genes involved in osteointegration and osteoinduction, such as OC, ALP, and COL1A1 in monocultures of osteoblasts and fibroblasts; co-cultures lacked this phenomenon. Immunocytochemistry and qPCR analysis showed slight upregulations of marker genes and corresponding proteins. This might be due to the potential stabilising effects of osteoblast-fibroblast cross talk in the co-culture environment, resembling fibrocartilage formation at the tendon-to-bone interface.

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