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Journal of Histochemistry and Cytochemistry, Vol. 49, 1165-1176, September 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

ADAM-10 Protein Is Present in Human Articular Cartilage Primarily in the Membrane-bound Form and Is Upregulated in Osteoarthritis and in Response to IL-1{alpha} in Bovine Nasal Cartilage

Susan Chubinskayaa,b, Rita Mikhaila, Angela Deutschc, and Michael H. Tindalc
a Department of Biochemistry, Health Care Research Center, Mason, Ohio
b Section of Rheumatology, Health Care Research Center, Mason, Ohio
c Rush Medical College, Rush-Presbyterian–St Luke's Medical Center, Chicago, Illinois, and Procter & Gamble Pharmaceuticals, Inc.,, Health Care Research Center, Mason, Ohio

Correspondence to: Susan Chubinskaya, Dept. of Biochemistry, Rush Medical College at Rush-Presbyterian–St Luke's Medical Center, 1653 W. Congress Parkway, Chicago, IL 60612.

The objective of our study was to determine the tissue distribution and localization of ADAM-10 protein in human and bovine cartilage and the changes it undergoes with cartilage degeneration seen in osteoarthritis (OA) and under the influence of interleukin-1 (IL-1). Human normal and OA articular cartilage and bovine nasal cartilage cultured in the presence of IL-1{alpha} were processed for histology and immunohistochemistry. ADAM-10 protein was extracted from human cartilage and analyzed by Western blotting using anti-ADAM-10 antibodies. Fluor S Image analyzer and Quantity One software program were applied to quantify the total amount of ADAM-10. ADAM-10 protein was detected in both human and bovine cartilage. The strongest immunostaining was found in the cytoplasm and/or cell membranes of the superficial and upper middle layer of normal adult human cartilage, in the clusters and fibrillated areas of OA cartilage, and in IL-1{alpha}-stimulated bovine nasal cartilage. The distribution of ADAM-10 protein in bovine nasal cartilage was dependent on the length of exposure to IL-1{alpha} and corresponded to the areas of proteoglycan depletion. By Western blotting analysis of human cartilage, ADAM-10 was primarily detected in the membrane-enriched fraction and its levels were increased in degenerated and OA cartilage compared to normal cartilage. The results of this study suggest that ADAM-10 might be an important factor associated with cartilage degenerative processes. (J Histochem Cytochem 49:1165–1176, 2001)

Key Words: human articular cartilage, bovine nasal cartilage, osteoarthritis, ADAM-10 protein, interleukin-1, immunohistochemistry


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