Quantitative image analysis is a form of imaging that includes microscopic histological quantification, video microscopy, image analysis, and image processing. Hallmarks are the generation of reliable, reproducible, and efficient measurements via strict calibration and s ...

In rheumatoid arthritis (RA), the synovium represents the predominant site of inflammation and joint destruction and is regarded as the key organ involved in disease pathogenesis. It has been studied in different ways over the last 30 yr, yielding information about the mechanisms involved ...

Necrosis and apoptosis have been demonstrated in articular cartilage in response to trauma and disease. However, cell death in articular cartilage may also be thought of as a scale of cell death culminating in secondary necrosis with the failure to remove apoptotic cells from the tissue. The in s ...

Assay of collagenase activity involves the use of radiolabeled collagen. Stimulation of cartilage with proinflammatory cytokines results in the upregulation of collagenases and the subsequent release of degraded collagen fragments. These enzymes can be localized in both oste ...

Quantitation of glycosaminoglycans (GAGs) in the form of aggrecan fragments released from cartilage in culture is a simple way to determine the efficacy of different cytokines alone or in combination in simulating cartilage catabolism. Two approaches for GAG assay are described, with ...

Two methods for the assessment of the expression of gelatinases A and B, MMP-2 and MMP-9, in articular cartilage are described. Immunohistochemical analysis of tissue sections provides information about the precise localization of the enzymes within the tissue, pinpointing the cells t ...

Membrane-type 1 matrix metalloproteinase (MT1-MMP) is a type I transmembrane protein which exhibits biological activities on the cell surface. One of the characteristic functions of this enzyme is activation of proMMP-2 on the cell surface (1). This process can be monitored using gelatin ...

This chapter provides practical information on the assay of tissue inhibitor of metalloproteinase (TIMP) activity and background information enabling meaningful interpretation of the data. Protocols are given for assessing the presence of TIMPs in biological samples by immun ...

Osteoclasts are bone-resorbing multinucleated cells derived from the monocytemacrophage lineage. The authors have developed a mouse marrow culture system and a coculture system of mouse osteoblasts and hemopoietic cells, in which osteoclasts are formed in response to various o ...

Rodent arthritis models are tools to study joint destruction. This chapter describes methods to prepare and analyze histological sections from inflamed paws. Protocols to create decalcified paraffin-embedded as well as undecalcified plastic-embedded sections are provide ...

In rheumatology and joint research, as in other fields, a purely descriptional appqoach to morphology cannot satisfy the exactions of modern clinical medicine. Investigators now appreciate the need to gauge pathological changes and their response to treatment by quantifying susc ...

The in vitro isolation, propagation and study of endothelial cells (EC) is an invaluable means by which the function of the vascular endothelium in physiology and pathophysiology can be explored. In recent years heterogeneity between large and small vessel EC, between arteries and veins, a ...

A complete pattern of adhesion molecules and chemokines involved in leukocyte migration in different tissues and in homeostatic vs inflammatory conditions is still lacking. This chapter describes how to characterize the mechanisms of leukocyte recruitment in synovial vessels ...

To be able to visualize real time leukocyte — endothelial cell interactions in vitro opens up the possibility of exploring the complex cascade of events that culminate in leukocyte recruitment and diapedesis in a much more detailed and controlled way. Techniques have been developed where ...

Angiogenesis is the formation of new blood vessels from existing vessels. The formation of new vessels appears to be an early and fundamental process for the evolution of the inflammatory response in synovial joints affected by arthritis. The propagation of new vessels in the synovial membr ...

The study of fibroblast-like synoviocytes (FLS) has yielded important insights into the pathogenic mechanisms of rheumatoid arthritis. FLS can be cultured from synovial tissue obtained at joint replacement surgery, synovectomy, or synovial biopsy. After collagenase digesti ...

The ingress of leukocytes into the synovium is a key event in inflammatory arthritis, such as rheumatoid arthritis. A number of soluble inflammatory mediators, such as chemokines, cytokines, and soluble adhesion molecules are involved in this event. It is feasible to test the chemotactic ac ...

Evaluation of differentially regulated genes is essential for the development of novel therapeutic approaches in multifactorial diseases such as rheumatoid arthritis (RA). RA synovial fibroblasts (RASF) are key players in inflammation and cartilage destruction. Therefor ...

The use of gene transfer techniques has become of utmost importance both for the analysis of molecular pathways of rheumatic joint destruction and for the evaluation of novel therapeutic concepts to treat rheumatic diseases. However, gene transfer into synovial fibroblasts faces sev ...

Rheumatoid arthritis is characterized by inflammation of the joints and degradation and invasion by fibroblast-like synoviocytes (FLS) of the cartilage. To assess the invasiveness of FLS an in vitro invasion assay was developed. In this invasion assay the FLS grow through an artificial m ...