Mechanical Strain Stimulates Osteoblast Proliferation Through the Estrogen Receptor in Males as Well as Females

Abstract

Mechanical strain, testosterone, and estrogen all stimulate proliferation of primary cultures of male rat long bone (LOB)‐derived osteoblast‐like cells as determined by [³H]thymidine incorporation. The maximum proliferative effect of a single period of mechanical strain (3400 με, 1 Hz, and 600 cycles) is additional to that of testosterone (10⁻⁸ M) or estrogen (10⁻⁸ M). The cells' proliferative response to strain is abolished both by concentrations of tamoxifen that cause proliferation (10⁻⁸ M) and by those that have no effect (10⁻⁶ M). Strain‐related proliferation also is reduced by the estrogen antagonist ICI 182,780 (10⁻⁸ M) but is unaffected by the androgen receptor antagonist hydroxyflutamide (10⁻⁷ M). Tamoxifen, ICI 182,780, and the aromatase inhibitor 4‐dihydroandrostenedione, at concentrations that have no effect on basal proliferation, significantly reduce the proliferative effect of the aromatizable androgen testosterone but not that of the nonaromatizable androgen 5α‐dihydrotestosterone. Hydroxyflutamide, at a concentration that has no effect on basal proliferation (10⁻⁷ M), eliminates the proliferative effect of 5α‐dihydro‐testosterone but had no significant effect on that caused by testosterone. Proliferation associated with strain is blocked by neutralizing antibody to insulin‐like growth factor II (IGF‐II) but not by antibody to IGF‐I. Proliferation associated with testosterone is blocked by neutralizing antibody to IGF‐I but is unaffected by antibody to IGF‐II. These data suggest that in rat osteoblast‐like cells from males, as from females, strain‐related proliferation is mediated through the estrogen receptor (ER) in a manner that does not compete with estrogen but that can be blocked by ER modulators. Proliferation associated with testosterone appears to follow its aromatization to estrogen and is mediated through the ER, whereas proliferation associated with 5α‐dihydrotestosterone is mediated by the androgen receptor. Strain‐related proliferation in males, as in females, is mediated by IGF‐II, whereas proliferation associated with estrogen and testosterone is mediated by IGF‐I.