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J. Biochem, 2004, Vol. 135, No. 5 567-575
© 2004 The Japanese Biochemical Society

CELL

Induction of CD44 and MMP Expression by Hyaluronidase Treatment of Articular Chondrocytes

Maiko Ohno-Nakahara*,1,2, Kobun Honda*,1, Kotaro Tanimoto1, Nobuaki Tanaka1, Takeyoshi Doi1, Aya Suzuki1, Kiyoshi Yoneno1, Yuki Nakatani1, Masashi Ueki1, Shigeru Ohno§,1,2, Warren Knudson2, Cheryl B. Knudson2 and Kazuo Tanne1

1 Department of Orthodontics and Craniofacial Developmental Biology, Hiroshima University Graduate School of Biomedical Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553; and 2 Department of Biochemistry, Rush Medical College, Rush University Medical Center, Chicago, IL 60612, USA

In this study, the effects of fragmentation of the glycosoaminoglycans of the cell-associated matrix by hyaluronidase (HAase) on the expression of CD44 receptor and matrix metalloproteinase (MMP) mRNAs in cultured articular chondrocytes were examined. Chondrocytes, isolated from rabbit and bovine articular cartilage, were treated with bovine testicular HAase (0–200 units/ml) in the presence or absence of an antibody for CD44. The mRNA levels of CD44, CD44 variant (CD44v), MMPs (MMP-1, -3 and -9), and tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2) were determined by RT-PCR. The treatment of cultured chondrocytes with HAase resulted in the production of low molecular weight fragments of hyaluronan (HA). The expression of CD44, CD44v and MMP (MMP-1, -3 and -9) mRNAs, but not TIMP-1 or TIMP-2 mRNA, was up-regulated in the cultures treated with HAase, whereas this expression was not affected by treatment with purified HA of 1.0 x 105 Da. Furthermore, the induction of CD44 and MMPs on treatment with HAase was suppressed by an anti-CD44 antibody. The results suggest that the fragmentation of HA may lead to cartilage destruction in terms of the enhanced expression of MMPs as well as the upregulation of CD44.

* Maiko Ohno-Nakahara and Kobun Honda contributed equally to this work.

§ To whom correspondence should be addressed. Department of Orthodontics and Craniofacial Developmental Biology, Hiroshima University Graduate School of Biomedical Sciences, Tel: +81-82-257-5686, Fax: +81-82-257-5687, E-mail: shigebon{at}hiroshima-u.ac.jp


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