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Journal of Dental Research
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Biological

Induction of MMP-3 by Hyaluronan Oligosaccharides in Temporomandibular Joint Chondrocytes

S. Ohno1,2, M. Ohno-Nakahara1,2, C.B. Knudson1 and W. Knudson1,*

1 Department of Biochemistry, Rush Medical College, Rush University Medical Center, 1653 W. Congress Parkway, Chicago, IL 60612, USA; and
2 Department of Orthodontics and Craniofacial Developmental Biology, Division of Cervico-Gnathostomatology, Hiroshima University Graduate School of Biomedical Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan;

Correspondence: * corresponding author, wknudson{at}rush.edu

Low-molecular-weight hyaluronan (LMW-HA) is often increased in osteoarthritic joints; however, its biological function in cartilage has not been clarified. We hypothesize that LMW-HA causes the catabolic activation of chondrocytes through its interaction with CD44. Cartilage explants and chondrocytes, derived from bovine temporomandibular joints (TMJ), were examined for matrix loss and the expression of matrix metalloproteinase-3 (MMP-3) following treatment with hyaluronan oligosaccharides (HAoligos). Hyaluronan and CD44 were uniformly distributed throughout the fibrous and cartilaginous zones of the TMJ condyle. Treatment of cartilage explants with HAoligos resulted in cartilage matrix loss with increased secreted caseinolytic activity. HAoligos treatment of TMJ chondrocytes resulted in enhanced MMP-3 expression, whereas wash-out of the HAoligos in the middle of the experimental period reduced this induction. These results suggest that HAoligos activate chondrocytes, resulting in a substantial enhancement of proteinase expression, and the removal of HAoligos by wash-out reverses this catabolic activation.

Key Words: temporomandibular joint • chondrocytes • hyaluronan • hyaluronan oligosaccharides • matrix metalloproteinase-3.

Abbreviations: mRNA, messenger ribonucleic acid • hr, hour • min, minute • DNA, deoxyribonucleic acid • SDS, sodium dodecyl sulfate • PAGE, polyacrylamide gel electrophoresis • GAPDH, glyceraldehyde-3-phosphate dehydrogenase • IgG, immunoglobulin G • HA, hyaluronan • LMW-HA, low-molecular-weight hyaluronan • HMW-HA, high-molecular-weight hyaluronan • HAoligos, hyaluronan oligosaccharides • HA6, hyaluronan hexasaccharides • OA, osteoarthritis • TMJ, temporomandibular joint • MMP-3, metalloproteinase-3 • RT-PCR, reverse-transcriptase/polymerase chain-reaction • DMEM, Dulbecco’s modified Eagle’s medium • FBS, fetal bovine serum • DAPI, 4', 6-diamidino-2-phenylindole, dihydrochloride • CP, cycle number at the crossing point.

Journal of Dental Research, Vol. 84, No. 11, 1005-1009 (2005)
DOI: 10.1177/154405910508401107
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