Immobilization causes rapid and massive bone loss. By comparing Botulinum Toxin A (BTX)-induced bone loss in mouse strains with different genetic backgrounds we investigated whether the genetic background had an influence on the severity of the osteopenia. Secondly, we investigated whether BTX had systemic effects on bone. Female mice from four inbred mouse strains (BALB/cJ, C57BL/6 J, DBA/2 J, and C3H/HeN) were injected unilaterally with BTX (n = 10/group) or unilaterally with saline (n = 10/group). Mice were euthanized after 21 days, and the bone properties evaluated using μCT, DXA, bone histomorphometry, and mechanical testing. BTX resulted in substantially lower trabecular bone volume fraction (BV/TV) and trabecular thickness in all mouse strains. The deterioration of BV/TV was significantly greater in C57BL/6 J (- 57%) and DBA/2 J (- 60%) than in BALB/cJ (- 45%) and C3H/HeN (- 34%) mice. The loss of femoral neck fracture strength was significantly greater in C57BL/6 J (- 47%) and DBA/2 J (- 45%) than in C3H (- 25%) mice and likewise the loss of mid-femoral fracture strength was greater in C57BL/6 J (- 17%), DBA/2 J (- 12%), and BALB/cJ (- 9%) than in C3H/HeN (- 1%) mice, which were unaffected. Using high resolution μCT we found no evidence of a systemic effect on any of the microstructural parameters of the contralateral limb. Likewise, there was no evidence of a systemic effect on the bone strength in any mouse strain. We did, however, find a small systemic effect on aBMD in DBA/2 J and C3H/HeN mice. The present study shows that BTX-induced immobilization causes the greatest loss of cortical and trabecular bone in C57BL/6 J and DBA/2 J mice. A smaller loss of bone microstructure and fracture strength was seen in BALB/cJ mice, while the bone microstructure and fracture strength of C3H/HeN mice were markedly less affected. This indicates that BTX-induced loss of bone is mouse strain dependent. We found only minimal systemic effects of BTX.

Modeling and remodeling induce significant changes of bone structure and mechanical properties with age. Therefore, it is important to gain knowledge of the processes taking place in bone over time. The rat is a widely used animal model, where much data has been accumulated on age-related changes of bone on the organ and tissue level, whereas features on the nano- and micrometer scale are much less explored. We investigated the age-related development of organ and tissue level bone properties such as bone volume, bone mineral density, and load to fracture and correlated these with osteocyte lacunar properties in rat cortical bone. Femora of 14 to 42-week-old female Wistar rats were investigated using multiple complementary techniques including X-ray micro-computed tomography and biomechanical testing. The body weight, femoral length, aBMD, load to fracture, tissue volume, bone volume, and tissue density were found to increase rapidly with age at 14-30 weeks. At the age of 30-42 weeks, the growth rate appeared to decrease. However, no accompanying changes were found in osteocyte lacunar properties such as lacunar volume, ellipsoidal radii, lacunar stretch, lacunar oblateness, or lacunar orientation with animal age. Hence, the evolution of organ and tissue level properties with age in rat cortical bone is not accompanied by related changes in osteocyte lacunar properties. This suggests that bone microstructure and bone matrix material properties and not the geometric properties of the osteocyte lacunar network are main determinants of the properties of the bone on larger length scales.

Thrombin is a key component in the coagulation cascade where it converts factor V, VIII, XI, and fibrinogen. In addition to the abundant production of thrombin in the liver, osteoclasts synthesize and secrete thrombin as well. Osteoblasts express thrombin receptors, and it has been reported that thrombin stimulates the expression of RANKL relatively to OPG, resulting in greater osteoclast activation and bone degradation. Pradaxa (dabigatran etexilate, DE) is a new anticoagulant, which has recently been approved for clinical use. DE is a direct thrombin inhibitor with potential to modulate the RANKL/OPG ratio and thereby limit osteoclast activation and bone degradation. The purpose of the study was to investigate whether DE can increase bone density, bone strength, and bone microstructure in healthy male and female mice and to investigate whether the effect of DE is sex-dependent. Twenty-eight 14-week-old male C57BL/6 mice were stratified by weight into 4 groups: 1. Control 3 weeks; 2. DE 3 weeks; 3. Control 6 weeks; 4. DE 6 weeks. An identical study design was applied to twenty-four 14-week-old female C57BL/6 mice. Chow mixed with DE was offered ad libitum, resulting in a dose of 1.70 mg DE/g body weight and 1.52 mg DE/g body weight, to female and male mice, respectively. The animals were euthanized after 3 or 6 weeks. Bone mineral density (aBMD) and bone mineral content (BMC) were evaluated with DEXA, 3D microstructural properties were determined with μCT, bone strength was determined with mechanical testing, and bone formation and resorption was evaluated with bone histomorphometry. In female mice, DE resulted in significant higher tibial aBMD values after 6 weeks of intervention. Furthermore, DE significantly increased tibial diaphyseal cortical bone area and tissue area, which was accompanied by significantly increased strength of the tibial shaft. DE had no effect on femoral cortical bone or on femoral and vertebral trabecular 3D microstructure. Finally, bone histomorphometry showed that DE had no effect on MS/BS or Oc.S/BS. In male mice, no bone positive effects of DE were found in any of the parameters investigated. In conclusion, intervention with DE may result in a weak positive site specific effect at tibial cortical bone in female mice, and importantly, no major deleterious effects of DE on bone tissue were seen in either female or male mice despite the relatively high dose of DE used.