scholarly journals The Expression of Thrombospondin-4 Correlates with Disease Severity in Osteoarthritic Knee Cartilage

2019 ◽  
Vol 20 (2) ◽  
pp. 447 ◽  
Author(s):  
Kathrin Maly ◽  
Inna Schaible ◽  
Jana Riegger ◽  
Rolf Brenner ◽  
Andrea Meurer ◽  
...  
Keyword(s):  
Protein Extraction ◽  
Degradation Products ◽  
Joint Disease ◽  
Transcriptional Level ◽  
Matrix Remodeling ◽  
Knee Cartilage ◽  
Cartilage Extracellular Matrix ◽  
Specific Degradation ◽  
Thrombospondin 4 ◽  
Extracellular Glycoprotein

Osteoarthritis (OA) is a progressive joint disease characterized by a continuous degradation of the cartilage extracellular matrix (ECM). The expression of the extracellular glycoprotein thrombospondin-4 (TSP-4) is known to be increased in injured tissues and involved in matrix remodeling, but its role in articular cartilage and, in particular, in OA remains elusive. In the present study, we analyzed the expression and localization of TSP-4 in healthy and OA knee cartilage by reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry, and immunoblot. We found that TSP-4 protein expression is increased in OA and that expression levels correlate with OA severity. TSP-4 was not regulated at the transcriptional level but we detected changes in the anchorage of TSP-4 in the altered ECM using sequential protein extraction. We were also able to detect pentameric and fragmented TSP-4 in the serum of both healthy controls and OA patients. Here, the total protein amount was not significantly different but we identified specific degradation products that were more abundant in sera of OA patients. Future studies will reveal if these fragments have the potential to serve as OA-specific biomarkers.

scholarly journals COMP and TSP-4: Functional Roles in Articular Cartilage and Relevance in Osteoarthritis

2021 ◽  
Vol 22 (5) ◽  
pp. 2242
Author(s):  
Kathrin Maly ◽  
Enrique Andres Sastre ◽  
Eric Farrell ◽  
Andrea Meurer ◽  
Frank Zaucke
Keyword(s):  
Articular Cartilage ◽  
Matrix Protein ◽  
Joint Disease ◽  
Chondrocyte Phenotype ◽  
Functional Roles ◽  
Ecm Proteins ◽  
Cartilage Extracellular Matrix ◽  
Thrombospondin 4 ◽  
Chondrocyte Migration

Osteoarthritis (OA) is a slow-progressing joint disease, leading to the degradation and remodeling of the cartilage extracellular matrix (ECM). The usually quiescent chondrocytes become reactivated and accumulate in cell clusters, become hypertrophic, and intensively produce not only degrading enzymes, but also ECM proteins, like the cartilage oligomeric matrix protein (COMP) and thrombospondin-4 (TSP-4). To date, the functional roles of these newly synthesized proteins in articular cartilage are still elusive. Therefore, we analyzed the involvement of both proteins in OA specific processes in in vitro studies, using porcine chondrocytes, isolated from femoral condyles. The effect of COMP and TSP-4 on chondrocyte migration was investigated in transwell assays and their potential to modulate the chondrocyte phenotype, protein synthesis and matrix formation by immunofluorescence staining and immunoblot. Our results demonstrate that COMP could attract chondrocytes and may contribute to a repopulation of damaged cartilage areas, while TSP-4 did not affect this process. In contrast, both proteins similarly promoted the synthesis and matrix formation of collagen II, IX, XII and proteoglycans, but inhibited that of collagen I and X, resulting in a stabilized chondrocyte phenotype. These data suggest that COMP and TSP-4 activate mechanisms to protect and repair the ECM in articular cartilage.

scholarly journals Pharmacological Targeting of Heme Oxygenase-1 in Osteoarthritis

Antioxidants ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 419
Author(s):  
Yohei Sanada ◽  
Sho Joseph Ozaki Tan ◽  
Nobuo Adachi ◽  
Shigeru Miyaki
Keyword(s):  
Heme Oxygenase ◽  
Redox Homeostasis ◽  
Synovial Fibroblasts ◽  
Protective Role ◽  
Joint Disease ◽  
Heme Oxygenase 1 ◽  
Transcriptional Level ◽  
Related Factor ◽  
Mobility Limitations ◽  
Therapeutic Implications

Osteoarthritis (OA) is a common aging-associated disease that clinically manifests as joint pain, mobility limitations, and compromised quality of life. Today, OA treatment is limited to pain management and joint arthroplasty at the later stages of disease progression. OA pathogenesis is predominantly mediated by oxidative damage to joint cartilage extracellular matrix and local cells such as chondrocytes, osteoclasts, osteoblasts, and synovial fibroblasts. Under normal conditions, cells prevent the accumulation of reactive oxygen species (ROS) under oxidatively stressful conditions through their adaptive cytoprotective mechanisms. Heme oxygenase-1 (HO-1) is an iron-dependent cytoprotective enzyme that functions as the inducible form of HO. HO-1 and its metabolites carbon monoxide and biliverdin contribute towards the maintenance of redox homeostasis. HO-1 expression is primarily regulated at the transcriptional level through transcriptional factor nuclear factor erythroid 2 (NF-E2)-related factor 2 (Nrf2), specificity protein 1 (Sp1), transcriptional repressor BTB-and-CNC homology 1 (Bach1), and epigenetic regulation. Several studies report that HO-1 expression can be regulated using various antioxidative factors and chemical compounds, suggesting therapeutic implications in OA pathogenesis as well as in the wider context of joint disease. Here, we review the protective role of HO-1 in OA with a focus on the regulatory mechanisms that mediate HO-1 activity.

scholarly journals In Arabidopsis thaliana mitochondria 5′ end polymorphisms of nad4L-atp4 and nad3-rps12 transcripts are linked to RNA PROCESSING FACTORs 1 and 8

2021 ◽  
Author(s):  
Sarah Schleicher ◽  
Stefan Binder
Keyword(s):  
Arabidopsis Thaliana ◽  
Rna Processing ◽  
Degradation Products ◽  
Underlying Mechanism ◽  
Transcriptional Level ◽  
Pentatricopeptide Repeat ◽  
Repeat Proteins ◽  
P Type ◽  
Pentatricopeptide Repeat Proteins ◽  
Processing Factors

Abstract Key message RNA PROCESSING FACTORs 1 AND 8 (RPF1 and RPF8), both restorer of fertility like pentatricopeptide repeat proteins, are required for processing of dicistronic nad4L-atp4 and nad3-rps12 transcripts in Arabidopsis mitochondria. Abstract In mitochondria of Arabidopsis thaliana (Arabidopsis), the 5′ termini of many RNAs are generated on the post-transcriptional level. This process is still poorly understood in terms of both the underlying mechanism as well as proteins required. Our studies now link the generation of polymorphic 5′ extremities of the dicistronic nad3-rps12 and nad4L-atp4 transcripts to the function of the P-type pentatricopeptide repeat proteins RNA PROCESSING FACTORs 8 (RPF8) and 1 (RPF1). RPF8 is required to generate the nad3-rps12 -141 5′ end in ecotype Van-0 whereas the RPF8 allele in Col has no function in the generation of any 5′ terminus of this transcript. This observation strongly suggests the involvement of an additional factor in the generation of the -229 5′ end of nad3-rps12 transcripts in Col. RPF1, previously found to be necessary for the generation of the -228 5′ end of the major 1538 nucleotide-long nad4 mRNAs, is also important for the formation of nad4L-atp4 transcripts with a 5′ end at position -318 in Col. Many Arabidopsis ecotypes contain inactive RPF1 alleles resulting in the accumulation of various low abundant nad4L-atp4 RNAs which might represent precursor and/or degradation products. Some of these ecotypes accumulate major, but slightly smaller RNA species. The introduction of RPF1 into these lines not only establishes the formation of the major nad4L-atp4 dicistronic mRNA with the -318 5′ terminus, the presence of this gene also suppresses the accumulation of most alternative nad4L-atp4 RNAs. Beside RPF1, several other factors contribute to nad4L-atp4 transcript formation.

scholarly journals USP7 Inhibition Alleviates H2O2-Induced Injury in Chondrocytes via Inhibiting NOX4/NLRP3 Pathway

2021 ◽  
Vol 11 ◽  
Author(s):  
Gang Liu ◽  
Qingbai Liu ◽  
Bin Yan ◽  
Ziqiang Zhu ◽  
Yaozeng Xu
Keyword(s):  
Treatment Options ◽  
Current Treatment ◽  
Pathological Process ◽  
Joint Disease ◽  
Matrix Remodeling ◽  
Ros Production ◽  
Caspase 1 ◽  
Enhanced Production

Osteoarthritis (OA), the most common form of arthritis, is a very common joint disease that often affects middle-aged to elderly people. However, current treatment options for OA are predominantly palliative. Thus, understanding its pathological process and exploring its potential therapeutic approaches are of great importance. Rat chondrocytes were isolated and exposed to hydrogen peroxide (H2O2) to mimic OA. The effects of H2O2 on ubiquitin-specific protease 7 (USP7) expression, reactive oxygen species (ROS) levels, proliferation, inflammatory cytokine release, and pyroptosis were measured. USP7 was knocked down (KD) or overexpressed to investigate the role of USP7 in OA. Co-immunoprecipitation (Co-IP) was used to study the interaction between USP7 and NAD(P)H oxidases (NOX)4 as well as NOX4 ubiquitination. NOX4 inhibitor was applied to study the involvement of NOX4 in USP7-mediated OA development. USP7 inhibitor was given to OA animals to further investigate the role of USP7 in OA in vivo. Moreover, H2O2 treatment significantly increased USP7 expression, enhanced ROS levels, and inhibited proliferation in rat chondrocytes. The overexpression of USP7 enhanced pyroptosis, ROS production, interleukin (IL)-1β and IL-18 levels, and the expression level of NLRP3, GSDMD-N, active caspase-1, pro-caspase-1, matrix metalloproteinases (MMP) 1, and MMP13, which was abolished by ROS inhibition. The USP7 KD protected rat chondrocytes against H2O2-induced injury. Co-IP results showed that USP7 interacted with NOX4, and USP7 KD enhanced NOX4 ubiquitinylation. The inhibition of NOX4 blocked the pro-OA effect of USP7. Moreover, the USP7 inhibitor given to OA animals suppressed OA in vivo. USP7 inhibited NOX4 ubiquitination for degradation which leads to elevated ROS production. ROS subsequently activates NLPR3 inflammasome, leading to enhanced production of IL-1β and IL-18, GSDMD-N-dependent pyroptosis, and extracellular matrix remodeling. Thus, UPS7 contributes to the progression of OA via NOX4/ROS/NLPR3 axis.

THE ROLE OF VARIOUS HAEMOSTASIS PARAMETERS IN MONITORING FIBRINOLYSIS WITH PRO-UROKINASE/STREPTOKINASE IN TREATMENT OF MYOCARDIAL INFARCTION

1987 ◽  
Author(s):  
F Keller ◽  
P Schanzenbächer ◽  
F Dati ◽  
J Huber ◽  
K kochsiek
Keyword(s):  
Myocardial Infarction ◽  
Degradation Products ◽  
Therapy Monitoring ◽  
Latex Agglutination ◽  
Systemic Activity ◽  
Thromboplastic Activity ◽  
Specific Degradation ◽  
D Dimer

The new drug pro-urokinase, a proenzyme of urokinase (scu-PA), seems to have advantages in comparison with other fibrinolytic agents. Properties like higher fibrin specifity, non-systemic activity and lower antigenity may lead to a lower rate of complications. In a pilot study 10 patients with acute myocardial infarction have been treated under angiographical control with pro-urokinase (3-9 millions IU) by i.v. application. In case of no perfusion a further administration of streptokinase was carried on. The blood samples were obtained at therapy begin and after 5, 10, 30, 60 and 120 minutes. The therapy monitoring was performed by determination of established haemostasis parameters, like fibrinogen, fibrin(ogen)-split products (FSP), a2-antiplasmin. Plasminogen and batroxobin-time. Furthermore, the diagnostic relevance of new laboratory tests for fibrinolysis, D-Dimer and thrombin-anti thrombin Ill-complex (TAT) has been investigated considering some typical follow-ups. D-Dimer were determined by latex agglutination test and TAT by enzyme immunoassay.Generally the application of pro-urokinase in contrast to streptokinase results in minimal changes of the classic fibrinolysis parameters like fibrinogen, FSP, batroxobin-time etc. demonstrating no systemic lysis. The appearance of plasmic degradation products of cross-linked fibrin (D-Dimer) is a specific indi-cater of the release of thrombotic material. Other non-specific degradation products (fibrinogenolysis) were detected by the measurement of FSP. In some cases in which perfusion ocurred an increase of TAT followed by a rapid decrease was observed. This indicates a higher thromboplastic activity which may originate from the infarcted area producing TAT complex formation.

scholarly journals Altered Integration of Matrilin-3 into Cartilage Extracellular Matrix in the Absence of Collagen IX

2005 ◽  
Vol 25 (23) ◽  
pp. 10465-10478 ◽  
Author(s):  
Bastian Budde ◽  
Katrin Blumbach ◽  
Joni Ylöstalo ◽  
Frank Zaucke ◽  
Harald W. A. Ehlen ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Knockout Mice ◽  
Matrix Protein ◽  
Protein Extraction ◽  
Direct Interaction ◽  
Matrix Proteins ◽  
Functional Protein ◽  
Cartilage Extracellular Matrix ◽  
Vertebral Bodies ◽  
Collagen Ix

ABSTRACT The matrilins are a family of four noncollagenous oligomeric extracellular matrix proteins with a modular structure. Matrilins can act as adapters which bridge different macromolecular networks. We therefore investigated the effect of collagen IX deficiency on matrilin-3 integration into cartilage tissues. Mice harboring a deleted Col9a1 gene lack synthesis of a functional protein and produce cartilage fibrils completely devoid of collagen IX. Newborn collagen IX knockout mice exhibited significantly decreased matrilin-3 and cartilage oligomeric matrix protein (COMP) signals, particularly in the cartilage primordium of vertebral bodies and ribs. In the absence of collagen IX, a substantial amount of matrilin-3 is released into the medium of cultured chondrocytes instead of being integrated into the cell layer as in wild-type and COMP-deficient cells. Gene expression of matrilin-3 is not affected in the absence of collagen IX, but protein extraction from cartilage is greatly facilitated. Matrilin-3 interacts with collagen IX-containing cartilage fibrils, while fibrils from collagen IX knockout mice lack matrilin-3, and COMP-deficient fibrils exhibit an intermediate integration. In summary, the integration of matrilin-3 into cartilage fibrils occurs both by a direct interaction with collagen IX and indirectly with COMP serving as an adapter. Matrilin-3 can be considered as an interface component, capable of interconnecting macromolecular networks and mediating interactions between cartilage fibrils and the extrafibrillar matrix.

Incorporation of 13C-Labeled Coniferyl Alcohol into Developing Ginkgo biloba L. Lignin Revealed by Analytical Pyrolysis and CuO Oxidation in Combination with Isotope Ratio Monitoring-Gas Chromatography-Mass Spectrometry

Holzforschung ◽  
2000 ◽  
Vol 54 (1) ◽  
pp. 39-54 ◽  
Author(s):  
T. I. Eglinton ◽  
M. A. Goñi ◽  
J. J. Boon ◽  
E. R. E. van der Hage ◽  
N. Terashima ◽  
...  
Keyword(s):  
Ginkgo Biloba ◽  
Degradation Products ◽  
Chemical Degradation ◽  
Original Position ◽  
Side Chain ◽  
Analytical Pyrolysis ◽  
Isotopic Analyses ◽  
Specific Degradation ◽  
Lignin Macromolecule ◽  
Cuo Oxidation

Summary A suite of four samples of xylem tissue from Ginkgo (Ginkgo biloba L.) shoots grown in a medium containing coniferin 13C-labeled at differing side-chain carbon atoms were studied using thermal and chemical degradation methods in combination with molecular-level isotopic analyses. The aims of the study were threefold: (1) to verify conclusions drawn from Nuclear Magnetic Resonance experiments previously performed on the same tissue samples, (2) to investigate degradation mechanisms and (3) to quantify the proportion of labeled material in each sample. Isotopic analysis of specific degradation products revealed the presence of the label exclusively within lignin-derived (phenolic) products and that the label is retained in its original position on the side-chain. These two results clearly indicate that there is no “scrambling” of carbon atoms as a result of thermal or chemical degradation, and thus lend strong support to analytical pyrolysis and chemolysis as viable approaches for structural investigations of the lignin macromolecule. Indeed, the isotopic enrichment of specific degradation products provides new evidence for certain types of linkages within the lignin polymer. The distribution and isotopic composition of the degradation products also strongly suggest an origin from newly-formed lignin as opposed to DHP-type products or unreacted substrate. As such, the data provides added confidence in the selective labeling approach for elucidation of the structure and biosynthesis of lignin. Isotopic mass balance calculations reveal that certain pyrolysis and CuO oxidation products show enhanced labeling which may be indicative of preferential incorporation of their specific precursors into the growing lignin macromolecule or heterogeneous lignin deposition.

scholarly journals Increased Knee Cartilage Volume in Degenerative Joint Disease using Percutaneously Implanted, Autologous Mesenchymal Stem Cells

2008 ◽  
Vol 3;11 (5;3) ◽  
pp. 343-353
Author(s):  
Dan Busse
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Pain Management ◽  
Degenerative Joint Disease ◽  
Joint Disease ◽  
Knee Cartilage ◽  
Pain Scores ◽  
Interventional Pain Management ◽  
Autologous Mesenchymal Stem Cells

Background: The ability to repair tissue via percutaneous means may allow interventional pain physicians to manage a wide variety of diseases including peripheral joint injuries and osteoarthritis. This review will highlight the developments in cellular medicine that may soon permit interventional pain management physicians to treat a much wider variety of clinical conditions and highlight an interventional case study using these technologies Objective: To determine if isolated and expanded human autologous mesenchymal stem cells could effectively regenerate cartilage and meniscal tissue when percutaneously injected into knees. Design: Case Study Setting: Private Interventional Pain Management practice. Methods: An IRB approved study with a consenting volunteer in which mesenchymal stem cells were isolated and cultured ex-vivo from bone marrow aspiration of the iliac crest. The mesenchymal stem cells were then percutaneously injected into the subject’s knee with MRI proven degenerative joint disease. Preand post-treatment subjective visual analog pain scores, physical therapy assessments, and MRIs measured clinical and radiographic changes. Results: At 24 weeks post-injection, the patient had statistically significant cartilage and meniscus growth on MRI, as well as increased range of motion and decreased modified VAS pain scores. Conclusion: The described process of autologous mesenchymal stem cell culture and percutaneous injection into a knee with symptomatic and radiographic degenerative joint disease resulted in significant cartilage growth, decreased pain and increased joint mobility in this patient. This has significant future implications for minimally invasive treatment of osteoarthritis and meniscal injury. Key words: autologous mesenchymal stem cells, osteoarthritis, knee, cartilage

scholarly journals The ECM-Cell Interaction of Cartilage Extracellular Matrix on Chondrocytes

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Yue Gao ◽  
Shuyun Liu ◽  
Jingxiang Huang ◽  
Weimin Guo ◽  
Jifeng Chen ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Large Family ◽  
Type Ii Collagen ◽  
Cell Interaction ◽  
Matrix Remodeling ◽  
Cell Matrix ◽  
Kinase C ◽  
Cartilage Extracellular Matrix ◽  
Adhesive Interactions ◽  
Wnt Signal

Cartilage extracellular matrix (ECM) is composed primarily of the network type II collagen (COLII) and an interlocking mesh of fibrous proteins and proteoglycans (PGs), hyaluronic acid (HA), and chondroitin sulfate (CS). Articular cartilage ECM plays a crucial role in regulating chondrocyte metabolism and functions, such as organized cytoskeleton through integrin-mediated signaling via cell-matrix interaction. Cell signaling through integrins regulates several chondrocyte functions, including differentiation, metabolism, matrix remodeling, responses to mechanical stimulation, and cell survival. The major signaling pathways that regulate chondrogenesis have been identified as wnt signal, nitric oxide (NO) signal, protein kinase C (PKC), and retinoic acid (RA) signal. Integrins are a large family of molecules that are central regulators in multicellular biology. They orchestrate cell-cell and cell-matrix adhesive interactions from embryonic development to mature tissue function. In this review, we emphasize the signaling molecule effect and the biomechanics effect of cartilage ECM on chondrogenesis.

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