Embryonic stem cell therapy improves bone quality in a model of impaired fracture healing in the mouse; tracked temporally using in vivo micro-CT.

In the current study, we used an estrogen-deficient mouse model of osteoporosis to test the efficacy of a cell-generated bone tissue construct for bone augmentation of an impaired healing fracture. A reduction in new bone formation at the defect site was observed in ovariectomized fractures compared to the control group using repeated measures in vivo micro-computed tomography (µCT) imaging over 4 weeks. A significant increase in the bone mineral density (BMD), trabecular bone volume ratio, and trabecular number, thickness and connectivity were associated with fracture repair in the control group, whereas the fractured bones of the ovariectomized mice exhibited a loss in all of these parameters (p<0.001). In a separate group, ovariectomized fractures were treated with murine embryonic stem (ES) cell-derived osteoblasts loaded in a three-dimensional collagen I gel and recovery of the bone at the defect site was observed. A significant increase in the trabecular bone volume ratio (p<0.001) and trabecular number (p<0.01) was observed by 4 weeks in the fractures treated with cell-loaded collagen matrix compared to those treated with collagen I alone. The stem cell-derived osteoblasts were identified at the fracture site at 4 weeks post-implantation through in situ hybridization histochemistry. Although this cell tracking method was effective, the formation of an ectopic cellular nodule adjacent to the knee joints of two mice suggested that alternative in vivo cell tracking methods should be employed in order to definitively assess migration of the implanted cells. To our knowledge, this study is the first of its kind to examine the efficacy of stem cell therapy for fracture repair in an osteoporosis-related fracture model in vivo. The findings presented provide novel insight into the use of stem cell therapies for bone injuries.

Signs of irreversible architectural changes occur early in the development of experimental osteoporosis as assessed by in vivo micro-CT.

UNLABELLED: Using in vivo micro-computed tomography, we assessed bone loss in the rat during the first twelve weeks after ovariectomy when structural changes were most rapid. Significant changes to the trabecular architecture were observed, including irreversible changes reflected by reduction in connectivity after only two weeks. This highlights that topological changes to the structure occur early in this experimental model of osteoporosis. INTRODUCTION: The purpose of this study was to establish a longitudinal time course of bone loss in the ovariectomized (OVX) rat model during the initial twelve-week period where structural changes are most rapid, and to identify when irreversible changes occur using in vivo micro-computed tomography (micro-CT). METHODS: The proximal tibiae of OVX (N = 10) and sham (N = 10) operated mature female Wistar rats were micro-CT scanned every two weeks from week 0 to week 12, excluding week 10. Changes in architecture were quantified using direct three-dimensional techniques and serum osteocalcin and CTX-I was measured at weeks 0, 6 and 12. Biomechanical properties were determined from femoral three-point bending and L-4 vertebral compression at the end of the protocol. ANOVA and paired t-tests were used to analyze the longitudinal and endpoint data, respectively. RESULTS: All of the measured architectural parameters changed significantly over the study in the OVX group, including irreversible changes reflected by connectivity density after two weeks. Osteocalcin concentration was elevated in the OVX group. Moderate changes in the mechanical properties of the femora midshaft and vertebrae were observed. CONCLUSIONS: Changes to the bone architecture and mechanics within twelve weeks after OVX highlight the importance of early diagnosis and treatment of osteoporosis.