Regulation of Matrix Metalloproteinases (MMP-2, -3, -9, and -13) by Interleukin-1 and Interleukin-6 in Mouse Calvaria: Association of MMP Induction with Bone Resorption

Abstract

Interleukin-1 (IL-1) greatly induces osteoclast formation and stimulates bone resorption of mouse calvaria in culture. In the presence of soluble IL-6 receptor (sIL-6R), IL-6 similarly induces osteoclast formation, but the potency of IL-6 in inducing bone resorption in organ culture is weaker than that of IL-1. To study the differences in bone-resorbing activity between IL-1 and IL-6, we examined the effects of the two cytokines on the induction of matrix metalloproteinases (MMPs). In mouse calvarial cultures, IL-1 markedly enhanced the messenger RNA (mRNA) expression of MMP-13 (collagenase 3), MMP-2 (gelatinase A), MMP-9 (gelatinase B), and MMP-3 (stromelysin 1), which associated with increases in bone matrix degradation. A hydroxamate inhibitor of MMPs significantly suppressed bone-resorbing activity induced by IL-1. Gelatin zymography showed that both pro- and active-forms of MMP-2 and MMP-9 were detected in the conditioned medium collected from calvarial cultures, and IL-1 markedly stimulated both pro- and active-forms of the two gelatinases. IL-6 with sIL-6R also stimulated mRNA expression and biological activities of these MMPs, but the potency was much weaker than that of IL-1. Conditioned medium collected from IL-1-treated calvariae degraded native type I collagen, but 3/4- and 1/4-length collagen fragments were not detected, suggesting that both collagenases and gelatinases synergistically degraded type I collagen into smaller fragments. In mouse osteoblastic cells, the expression ofMMP-2, MMP-3, and MMP-13 mRNAs could be detected, and they were markedly enhanced by IL-1alpha on days 2 and 5. IL-6 with sIL-6R also induced expression of MMP-13 and MMP-2 mRNAs on day 2, but the expression was rather transient. These results demonstrate that the potency of induction of MMPs by IL-1 and IL-6 is closely linked to the respective bone-resorbing activity, suggesting that MMP-dependent degradation of bone matrix plays a key role in bone resorption induced by these cytokines.

Activation of the receptor EphB4 by its specific ligand ephrin B2 in human osteoarthritic subchondral bone osteoblasts

“…Interestingly, osteoblasts treated with ephrin B2 significantly inhibited the expression of MMP-1, MMP-9, and MMP-13, but not MMP-2. This could have occurred through a direct effect of EphB4 receptor activation or through an indirect effect via inhibition of the cytokine IL-1␤, a major factor involved in the up-regulation of MMP in articular joint tissues (41). The inhibition of these MMPs is important, since these proteases are well-known for their induction of bone resorption via the degradation of extracellular matrix components, including the collagen fiber network, as well as other extracellular components, such as fibronectin and aggrecans.…”

Significant increases in serum and plasma concentrations of matrix metalloproteinases 3 and 9 in patients with rapidly destructive osteoarthritis of the hip

“…MMP-9 has been implicated in cellular migration and invasion in patients with such conditions as inflammation, tumor invasion, and metastasis. In addition, recent studies of this proteinase in the skeleton suggest that MMP-9 could exist in matrix vesicles (27), digest cartilage proteoglycans (28,29), and play important roles in chondrocyte apoptosis (30) and osteoclastic bone resorption (31)(32)(33). MMP-9 can also be produced by activated osteoclasts (34).…”

Role of RANKL-induced osteoclast formation and MMP-dependent matrix degradation in bone destruction by breast cancer metastasis

“…For Northern blotting, 20 mg of total RNA was resolved using electrophoresis on a 1% agarose -formaldehyde gel and transferred onto a nylon membrane (Hybond-N, Amersham Pharmacia Biotech, Tokyo, Japan), which was then hybridised with a 32 P-labelled cDNA probe, as reported previously . A 946-bp fragment of mouse RANKL cDNA and a 485-bp fragment of mouse MMP-13 were prepared using RT -PCR cloning and used as the probes (Kusano et al, 1998;Yasuda et al, 1998). Mouse MT1-MMP cDNA was kindly provided by Dr M Seiki, and a 656-bp fragment was used as the probe (Sato et al, 1994).…”

“…More than 20 different mammalian MMPs have been identified, and divided into four subgroups; collagenase (MMP-1, MMP-8, MMP-13 and MMP-18), gelatinase (MMP-2 and MMP-9), stromelysin (MMP-3 and MMP-10) and membrane-type metalloproteinase (MMPs-14 -17) (Mauviel, 1993;Nagase and Woessner, 1999;Vu and Werb, 2000). It is known that osteoblasts produce various MMPs such as MMP-1, MMP-2, MMP-13 (collagenase 3) and MMP-14 (MT1-MMP), and that osteoclasts selectively produce MMP-9 (MacDougall and Matrisian, 1995;Kusano et al, 1998;Westermarck and Kahari, 1999). We have reported that the induction of MMPs (such as MMP-2, -3 and -13) in osteoblasts is essential for bone resorption (Kusano et al, 1998).…”

“…It is known that osteoblasts produce various MMPs such as MMP-1, MMP-2, MMP-13 (collagenase 3) and MMP-14 (MT1-MMP), and that osteoclasts selectively produce MMP-9 (MacDougall and Matrisian, 1995;Kusano et al, 1998;Westermarck and Kahari, 1999). We have reported that the induction of MMPs (such as MMP-2, -3 and -13) in osteoblasts is essential for bone resorption (Kusano et al, 1998). On the other hand, the roles of MMPs in infiltration and metastasis of tumour cells have been widely investigated (MacDougall and Matrisian, 1995;Nakahara et al, 1997;Westermarck and Kahari, 1999).…”