Neoepitopes reveal the features of type II collagen cleavage and the identity of a collagenase involved in the transformation of the epiphyses anlagen in development

Lee, Eunice R.; Lamplugh, Lisa; Kluczyk, Beata; Leblond, C. P.; Mort, John S.

doi:10.1002/dvdy.21960

Cited by 18 publications

(9 citation statements)

References 81 publications

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“…Briefly, sections were pretreated with 0.25 units/ml of protease‐free chondroitinase ABC in phosphate buffered saline (PBS; Sigma‐Aldrich), and 1% hyaluronidase in 0.1 M Tris acetate (Sigma‐Aldrich) for 60 minutes at 37°C. The specimens were incubated for 18 hours at 4°C with the following primary antibodies: goat polyclonal anti‐EphB4 (1:100 dilution; R&D Systems), rabbit polyclonal anti–C‐terminal peptide of aggrecan G1 domain (VDIPEN, 1:800 dilution; provided by Dr. J. S. Mort, Shriners Hospital for Children, McGill University Hospital Centre, Montreal, Quebec, Canada) (32), a rabbit polyclonal antibody that represents a type II collagen primary cleavage site (Col2‐3/4C short , 1:200 dilution; provided by Dr. A. R. Poole, Shriners Hospital for Children, McGill University Hospital Centre, Montreal, Quebec, Canada) (33), and mouse anti–type X collagen antibody (1:100 dilution; provided by Dr. E. Lee, Shriners Hospital for Children, McGill University Hospital Centre, Montreal, Quebec, Canada) (34).…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, sections were pretreated with 0.25 units/ml of protease-free chondroitinase ABC in phosphate buffered saline (PBS; Sigma-Aldrich), and 1% hyaluronidase in 0.1M Tris acetate (Sigma-Aldrich) for 60 minutes at 37°C. The specimens were incubated for 18 hours at 4°C with the following primary antibodies: goat polyclonal anti-EphB4 (1:100 dilution; R&D Systems), rabbit polyclonal anti-Cterminal peptide of aggrecan G1 domain ( (34).…”

Section: Bone-specific Ephb4 Receptor-overexpressing Micementioning

confidence: 99%

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In vivo bone‐specific EphB4 overexpression in mice protects both subchondral bone and cartilage during osteoarthritis

Valverde-Franco

Pelletier

Fahmi

et al. 2012

Arthritis & Rheumatism

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Objective. In vitro activation of the receptor EphB4 positively affects human osteoarthritis (OA) articular cell metabolism. However, the specific in vivo role of this ephrin receptor in OA remains unknown. We investigated in mice the in vivo effect of bone-specific EphB4 overexpression on OA pathophysiology.Methods. Morphometric, morphologic, and radiologic evaluations were performed on postnatal day 5 (P5) mice and on 10-week-old mice. Knee OA was induced surgically by destabilization of the medial meniscus (DMM) in 10-week-old male EphB4 homozygous transgenic (EphB4-Tg) and wild-type (WT) mice. Medial compartment evaluations of cartilage were performed using histology and immunohistochemistry, and evaluations of subchondral bone using histomorphometry, osteoclast staining, and micro-computed tomography.Results. There was no obvious phenotype difference in skeletal development between EphB4-Tg mice and WT mice at P5 or at 10 weeks. At 8 and 12 weeks post-DMM, the EphB4-Tg mice demonstrated significantly less cartilage alteration in the medial tibial plateau and the femoral condyle than did the WT mice.This was associated with a significant reduction of aggrecan and type II collagen degradation products, type X collagen, and collagen fibril disorganization in the operated EphB4-Tg mice. The medial tibial subchondral bone demonstrated a significant reduction in sclerosis, bone volume, trabecular thickness, and number of tartrate-resistant acid phosphatase-positive osteoclasts at both times assessed post-DMM in the EphB4-Tg mice than in the WT mice.Conclusion. This is the first in vivo evidence that bone-specific EphB4 overexpression exerts a protective effect on OA joint structural changes. The findings of this study stress the in vivo importance of subchondral bone biology in cartilage integrity.Osteoarthritis (OA) is the most common form of arthritis and a leading cause of long-term disability. With increasing life expectancy, OA is a major socioeconomic and clinical concern, as no curative treatment yet exists. While considerable advancement has been made toward a better understanding of the pathophysiology of the disease process, there is still much to be accomplished before a disease-modifying OA drug is developed that can effectively reduce or stop the disease progression. It is therefore of the utmost importance to identify new candidates that can contribute to the development of therapeutic agents to prevent or arrest the disease process.Although the hallmark of OA is the progressive degeneration of articular cartilage, the subchondral bone is also suggested to be an active component of the OA process in humans (1-4). The rationale is that because the subchondral bone plate is in direct contact with the cartilage, it influences not only mechanical effects, but also cartilage degradation by providing catabolic factors to this overlying tissue, thus promoting Dr.

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Section: Methodsmentioning

confidence: 99%

Section: Bone-specific Ephb4 Receptor-overexpressing Micementioning

confidence: 99%

In vivo bone‐specific EphB4 overexpression in mice protects both subchondral bone and cartilage during osteoarthritis

Valverde-Franco

Pelletier

Fahmi

et al. 2012

Arthritis & Rheumatism

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“…[72] Another collagen-binding peptidel abeled with 99m Tc wasr eported recently. [73] Peptide CBP1495i sapeptide domain from pro-MMP-2, [74] which is ap recursor of the enzyme MMP-2 that interacts with type Ic ollagen. [75] The efficacy of probe 99m Tc-CBP1495 ( Figure 6B)w as evaluated in rat modelso fb leomycin-induced pulmonary fibrosis and CCl 4 -induced liver fibrosis.…”

Section: Pet/spectimagingmentioning

confidence: 99%

Molecular Probes for Imaging Fibrosis and Fibrogenesis

Désogère

Montesi

Caravan

2018

Chemistry A European J

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Fibrosis, or the accumulation of extracellular matrix molecules that make up scar tissue, is a common result of chronic tissue injury. Advances in the clinical management of fibrotic diseases have been hampered by the low sensitivity and specificity of noninvasive early diagnostic options, lack of surrogate end points for use in clinical trials, and a paucity of noninvasive tools to assess fibrotic disease activity longitudinally. Hence, the development of new methods to image fibrosis and fibrogenesis is a large unmet clinical need. Herein, an overview of recent and selected molecular probes for imaging of fibrosis and fibrogenesis by magnetic resonance imaging, positron emission tomography, and single photon emission computed tomography is provided.

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“…This can be used in a variety of biological applications. For example, an antibody against a neoepitope of collagen II has been used to assess the role of MMP13 in bone development (Lee, Lamplugh, Kluczyk, Leblond, & Mort, 2009). …”

Section: Advanced Imaging For Visualization Of Collagen and Other Ecmmentioning

confidence: 99%

Methods for the visualization and analysis of extracellular matrix protein structure and degradation

Léonard

Loughran

Klymenko

et al. 2018

Methods in Cell Biology

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This chapter highlights methods for visualization and analysis of extracellular matrix (ECM) proteins, with particular emphasis on collagen type I, the most abundant protein in mammals. Protocols described range from advanced imaging of complex in vivo matrices to simple biochemical analysis of individual ECM proteins. The first section of this chapter describes common methods to image ECM components and includes protocols for second harmonic generation, scanning electron microscopy, and several histological methods of ECM localization and degradation analysis, including immunohistochemistry, Trichrome staining, and in situ zymography. The second section of this chapter details both a common transwell invasion assay and a novel live imaging method to investigate cellular behavior with respect to collagen and other ECM proteins of interest. The final section consists of common electrophoresis-based biochemical methods that are used in analysis of ECM proteins. Use of the methods described herein will enable researchers to gain a greater understanding of the role of ECM structure and degradation in development and matrix-related diseases such as cancer and connective tissue disorders.

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Neoepitopes reveal the features of type II collagen cleavage and the identity of a collagenase involved in the transformation of the epiphyses anlagen in development

Cited by 18 publications

References 81 publications

In vivo bone‐specific EphB4 overexpression in mice protects both subchondral bone and cartilage during osteoarthritis

In vivo bone‐specific EphB4 overexpression in mice protects both subchondral bone and cartilage during osteoarthritis

Molecular Probes for Imaging Fibrosis and Fibrogenesis

Methods for the visualization and analysis of extracellular matrix protein structure and degradation

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