Preventive effects of the anti-vasospasm agent via the regulation of the Rho-kinase pathway on the development of steroid-induced osteonecrosis in rabbits.

A number of studies have suggested that ischemia is the principal pathomechanism of osteonecrosis, however, the detailed mechanism responsible for ischemia remains unclear. We examined the effects of fasudil, an anti-vasospasm agent, on the development of steroid-induced osteonecrosis in rabbits. One group of rabbits received 15mg/kg of fasudil intravenously, which were then injected once intramuscularly with 20mg/kg of methylprednisolone (n=33), and one received methylprednisolone alone as a control (n=28). Eight rabbits from each group were sacrificed 24h after methylprednisolone injection to analyze them by the expression of endothelinA-receptor and eNOS. Two weeks after the steroid injection, the femora and humeri were examined histopathologically for the incidence of osteonecrosis. In addition, plasma from each of four osteonecrosis-positive or -negative rabbits was used for the proteomic analysis in the fasudil group. The incidence of osteonecrosis was significantly lower in the fasudil group (32%) than that in the control group (75%) (P<0.01). Immunohistochemically, endothelinA-receptor expressions levels were decreased in the smooth muscle of the bone marrow in the fasudil group in comparison to that in the control group. The eNOS expressions levels in both serum and bone marrow in the MF group were significantly higher than those in the M group (P<0.05). Based on the proteomic analysis, several proteins related to vasospasm, such as fibrinogen, thrombin, and apolipoprotein E, were identified in rabbits with osteonecrosis soon after steroid administration. This study indicates that vasospasm is one of the important factors involved in the pathogenesis of steroid-induced osteonecrosis and that the anti-vasospasm agents seem to decrease the incidence of steroid-induced osteonecrosis.