Title:
Discordant recovery of bone mass and mechanical properties during prolonged recovery from disuse.

Abstract:
Profound bone loss at weight bearing sites is a primary effect of long-duration spaceflight. Moreover, a significant increase in estimated fracture risk remains even 1 year after returning to Earth; hence, it is important to define how quickly bone integrity can recover following prolonged disuse. This study characterized the loss and recovery dynamics of bone following a period of rodent hindlimb unloading in three anatomic sites. We hypothesized that the rat femoral neck would exhibit a discordant recovery dynamic most similar to that observed in astronauts' proximal femur; that is, bone mineral content (absolute mass) at this site would recover faster and more completely than would bone density and cortical area, and they will all recover before bone strength does. We characterized loss and long-term recovery of densitometric properties at the femoral neck, proximal tibia metaphysis, and tibia diaphysis, and also mechanical properties at the femoral neck and tibia diaphysis for which mechanical testing is amenable. We assessed the relationship between calculated strength indices and measured mechanical properties. Adult male Sprague-Dawley rats (6 months) were assigned to baseline, age-matched control (AC), and hindlimb unloaded (HU) groups. The HU group was unloaded for 28 days and then returned to normal cage activity for 84 days of weight bearing recovery (3 times the duration of HU). Fifteen animals were euthanized from each of the HU and AC groups on days 28, 56, 84, and 112 of the study. At baseline and then every 28 days in vivo longitudinal pQCT scans were taken at proximal tibia metaphysis (PTM) and tibia diaphysis (TD); ex vivo pQCT scans were taken later at the femoral neck (FN). TD and FN were tested to failure to measure mechanical properties. The hypothesis that the femoral neck in rats will exhibit a discordant recovery dynamic most similar to that observed in astronauts' proximal femurs was not supported by our data. At the femoral neck, densitometric and geometric variables (total BMC, total vBMD, cancellous vBMD, and cortical area) recovered to age-matched control levels after a recovery period twice the duration of unloading. Contrary to our hypothesis, changes in densitometric variables at the PTM provided a better model for changes in the human femoral neck with prolonged weightlessness. Following 28 days of HU, PTM total BMC recovered to age-matched control levels after roughly two times the duration of unloading; however, total vBMD did not recover even after three recovery periods. Cortical thinning occurred at the PTM following HU likely due to inhibition of periosteal growth; cortical shell thickness did not recover even after three recovery periods. Calculated strength indices suggested a loss in strength at the tibial diaphysis, which was not confirmed with direct testing of mechanical properties. HU had no effect on maximum fracture force at mid-tibia diaphysis; however, femoral neck experienced a significant loss of maximum force due to unloading that fully recovered after 28 days. Estimated strength indices for the femoral neck suggested a recovery period of 56 days in contrast to the 28-day recovery that was observed with mechanical testing. However, the inaccuracy of strength indices vs. directly measured mechanical properties highlights the continued importance of ground based animal models and mechanical testing. Our results demonstrate that the PTM in the rat better matches loss and recovery dynamics observed in astronauts' proximal femur than does the rat FN, at least in terms of densitometric variables. More complete utility of the rat PTM as a model in this case, however, depends upon meaningful characterization of changes in mechanical properties as well.