Stromelysin (MMP-3) Synthesis Is Up-Regulated in Estrogen-Deficient Mouse Osteoblasts In Vivo and In Vitro

Abstract

Sex steroids are important regulators of bone cell function and osteoblast‐derived matrix metalloproteinases (MMPs) are key mediators of bone resorption during the initial stage of osteoid removal prior to osteoclast attachment. To investigate the mechanism of bone loss following estrogen deficiency, we examined the effects of estrogen on osteoblast synthesis of MMPs and tissue inhibitor of metalloproteinases (TIMPs). Immunolocalization in mouse bone samples ex vivo and primary mouse osteoblast (MOB) cultures was used to document the synthesis of mouse interstitial collagenase (MMP‐13), stromelysin‐1 (MMP‐3), gelatinase‐A (MMP‐2), and gelatinase‐B (MMP‐9). Endosteal bone lining cells from distal femoral head and lumbar vertebral body showed an increase in the pattern of synthesis of stromelysin‐1 following ovariectomy, compared with sham‐operated controls; the synthesis of other MMPs was unaffected. The expression of all classes of MMPs and TIMP‐1 and TIMP‐2 by MOB in culture was demonstrated by reverse transcriptase‐polymerase chain reaction. Following the withdrawal of 17β‐estradiol, MOB cultures showed a significant increase in the number of cells synthesizing stromelysin‐1; this effect was enhanced by stimulation with either interleukin‐1 or interleukin‐6. Northern blot analysis showed only a slight increase in stromelysin‐1 mRNA message following the withdrawal of 17β‐estradiol. Our data show an unexpected up‐regulation of stromelysin‐1 synthesis by osteoblasts both in vivo and in vitro following estrogen withdrawal. Although this effect was not reflected in a significant change in stromelysin‐1 mRNA expression in vitro, there is evidence to suggest a role for this enzyme in the early stages of bone loss during the pathogenesis of osteoporosis.