Zoledronate reduces loading-induced microdamage in cortical ulna of ovariectomized rats.

As a daily physiological mechanism in bone, microdamage accumulation dissipates energy and helps to prevent fractures. However, excessive damage accumulation might bring adverse effects to bone mechanical properties, which is especially problematic among the osteoporotic and osteopenic patients treated by bisphosphonates. Some pre-clinical studies in the literature applied forelimb loading models to produce well-controlled microdamage in cortical bone. Ovariectomized animals were also extensively studied to assimilate human conditions of estrogen-related bone loss. In the present study, we combined both experimental models to investigate microdamage accumulation in the context of osteopenia and zoledronate treatment. Three-month-old normal and ovariectomized rats treated by saline or zoledronate underwent controlled compressive loading on their right forelimb to create in vivo microdamage, which was then quantified by barium sulfate contrast-enhanced micro-CT imaging. Weekly in vivo micro-CT scans were taken to evaluate bone (re)modeling and to capture microstructural changes over time. After sacrifice, three-point-bending tests were performed to assess bone mechanical properties. Results show that the zoledronate treatment can reduce cortical microdamage accumulation in ovariectomized rats, which might be explained by the enhancement of several bone structural properties such as ultimate force, yield force, cortical bone area and volume. The rats showed increased bone formation volume and surface after the generation of microdamage, especially for the normal and the ovariectomized groups. Woven bone formation was also observed in loaded ulnae, which was most significant in ovariectomized rats. Although all the rats showed strong correlations between periosteal bone formation and microdamage accumulation, the correlation levels were lower for the zoledronate-treated groups, potentially because of their lower levels of microdamage. The present study provides insights to further investigations of pharmaceutical treatments for osteoporosis and osteopenia. The same experimental concept can be applied in future studies on microdamage and drug testing.

Impact loading intensifies cortical bone (re)modeling and alters longitudinal bone growth of pubertal rats.

Physical exercise is important for musculoskeletal development during puberty, which builds bone mass foundation for later in life. However, strenuous levels of training might bring adverse effects to bone health, reducing longitudinal bone growth. Animal models with various levels of physical exercise were largely used to provide knowledge to clinical settings. Experiments from our previous studies applied different levels of mechanical loading on rat tibia during puberty accompanied by weekly in vivo micro-CT scans. In the present article, we apply 3D image registration-based methods to retrospectively analyze part of the previously acquired micro-CT data. Longitudinal bone growth, growth plate thickness, and cortical bone (re)modeling were evaluated from rats' age of 28-77 days. Our results show that impact loading inhibited proximal bone growth throughout puberty. We hypothesize that impact loading might bring different growth alterations to the distal and proximal growth plates. High impact loading might lead to pathological consequence of osteochondrosis and catch-up growth due to growth inhibition. Impact loading also increased cortical bone (re)modeling before and after the peak proximal bone growth period of young rats, of which the latter case might be caused by the shift from modeling to remodeling as the dominant activity toward the end of rat puberty. We confirm that the tibial endosteum is more mechano-sensitive than the periosteum in response to mechanical loading. To our knowledge, this is the first study to follow up bone growth and bone (re)modeling of young rats throughout the entire puberty with a weekly time interval.

Validation of an in vivo micro-CT-based method to quantify longitudinal bone growth of pubertal rats.

Bone growth is an essential part of skeletal development during childhood and puberty. Accurately characterizing longitudinal bone growth helps to better understand the determining factors of peak bone mass, which has impacts on bone quality later in life. Animal models were largely used to study longitudinal bone growth. However, the commonly used histology-based method is destructive and unable to follow up the growth curve of live animals in longitudinal experiments. In this study, we validated an in vivo micro-CT-based method against the histology-based method to quantify longitudinal bone growth rates of young rats non-destructively. CD (Sprague Dawley) IGS rats aged 35, 49 and 63 days received the same treatments: two series of repeated in vivo micro-CT scans on their proximal hind limb at a five-day interval, and two calcein injections separated by three days. The longitudinal bone growth rate was quantified by registering time-lapse micro-CT images in 3D, calculating the growth distance on registered images, and dividing the distance by the five-day gap. The growth rate was also evaluated by measuring the 2D distance between consecutive calcein fluorescent bands on microscopic images, divided by the three-day gap. The two methods were both validated independently with reproducible repeated measurements, where the micro-CT-based method showed higher precision. They were also validated against each other with low relative errors and a strong Pearson sample correlation coefficient (0.998), showing a significant (p < 0.0001) linear correlation between paired results. We conclude that the micro-CT-based method can serve as an alternative to the histology-based method for the quantification of longitudinal growth. Thanks to its non-invasive nature and true 3D capability, the micro-CT-based method helps to accommodate in vivo longitudinal animal studies with highly reproducible measurements.