Comparison of three methods for estimation of bone resorption following ovariectomy in the distal femur and the proximal tibia of the rat

The study of estrogen-deficient bone loss requires accurate assessment of bone resorption; however, there has been considerable variance in the values reported for this variable. We compared three techniques for estimating bone resorption in the adult rat at four anatomical sites, the metaphysis and epiphysis of the distal femur and the proximal tibia. The techniques include an osteoclast-morphology-based method (VK-Oc), a bone-surface-based method (Pit-Oc), and an enzymatic method (AP-Oc). Thirty 6-month-old rats were either ovariectomized (ovx) or sham operated (sham) and killed at 0, 9, or 18 days postoperation. Each method was analyzed for variance in the ovary-intact groups and for the ability to detect the increase in osteoclast surface known to occur following ovariectomy. A 50-fold variation for the estimation of extent of osteoclast surface was entirely accounted for by these three methods for osteoclast estimation. The VK-Oc method was the most consistent for discriminating levels of osteoclast surface between ovary-intact and ovariectomized rats, detecting an increase at three of the four sites. There was no difference between the methods in their ability to produce consistent values for ovary-intact groups. The Pit-Oc method produced the largest numerical difference between the two operation groups with a 2.5–3-fold increase compared with 1.25–1.5-fold for the other methods (p< 0.001). However, the greater variance associated with this method limited the ability to detect the increase in osteoclast surface following ovariectomy. The AP-Oc method lacked the sensitivity of the other two methods. The oophorectomy-induced increase of osteoclast surface in the metaphysis of distal femur was larger and more consistently demonstrated than increases at the other sites.