Temporal changes in bone mass and mechanical properties in a murine model of tumor osteolysis.

Pathological fracture is a devastating complication of osteolytic bone metastases. The progression of osteolysis and its effect on bone fracture risk are poorly understood. The goal of this study was to determine the temporal changes in bone strength following tumor inoculation in a preclinical model of tumor osteolysis. In addition, a predictive model was developed between non-invasive radiographic measures and bone strength. The right femora of female nude mice were injected with breast cancer cells; the left limb served as a sham-operated control. Radiographs and DEXA scans were obtained at the time of surgery and at 3, 6, and 9 weeks. Groups of mice were euthanized at each time point for mechanical assessment. Micro-CT analysis was performed on a sub-set of mice with advanced state disease to quantify bone loss. Radiographs documented an increase in tumor osteolysis over time, with 58% of the mice showing signs of osteolysis at 3 weeks, 75% at 6 weeks, and 81% at 9 weeks. BMD measurements revealed a 21.6% increase from baseline in the controls whereas tumor-injected femora failed to increase in BMD over the same time course. Tumor-bearing limbs exhibited statistically significant decreases in torque at failure (86%), energy to failure (88%), and initial stiffness (94%) compared to the controls. Both lysis scores and BMD measurements proved to be modest predictors of mechanical strength, accounting for approximately 73% and 41% of variation in torque at failure, respectively. Micro-CT analysis revealed decreases in both total bone volume in the distal femur (31%) and metaphyseal fractional trabecular bone (89%). We have shown that non-invasive radiographic techniques provide a useful tool for monitoring the progression of tumor osteolysis and for predicting the mechanical strength of tumor-bearing bones in this model. By integrating non-invasive measures of tumor osteolysis and fracture risk, we have validated a clinically relevant platform for evaluating new therapeutic approaches for preserving and/or restoring bone affected by metastatic disease.

Preclinical evaluation of a poly (vinyl alcohol) hydrogel implant as a replacement for the nucleus pulposus.

STUDY DESIGN: An in vivo investigation into the safety of a novel hydrogel implant designed to replace the diseased nucleus pulposus. OBJECTIVES: To determine the local and systemic safety of this new implant in a nonhuman primate model. SUMMARY OF BACKGROUND DATA: A poly (vinyl alcohol) (PVA) hydrogel has been developed as a prosthetic replacement for the diseased nucleus pulposus. METHODS: PVA implants were inserted into discectomy defects created in the L3-L4 or L4-L5 intervertebral disc in 20 male baboons. Empty discectomy defects served as a surgical control in 8 additional animals. Routine follow-up evaluations included radiography, magnetic resonance imaging, gross pathology, and histopathology of both local and remote tissues. RESULTS: Insertion of the PVA hydrogel from an anterior direction produced extrusions in 5 animals from the first series of 15 surgeries (33%). A modified surgical technique, involving an anterolateral rather than anterior approach, was used in 5 animals, but the extrusion rate remained high (20%). Despite these surgical complications, the PVA implants were well tolerated over 24 months in vivo, with no evidence of device-related pathology in the adjacent disc tissue, spinal cord, or remote tissues. CONCLUSION: Implantation of the PVA implant for periods of up to 24 months produced no evidence of local or systemic toxicity. Additional studies are now needed to determine the efficacy of the device in its intended application.

Defect repair in the rat mandible with bone morphogenic proteins and marrow cells.

OBJECTIVE: To investigate the ability of a bone growth factor mixture and bone marrow cells to repair a critical size defect of the rat mandibular body. DESIGN: Prospective, randomized controlled trial. SUBJECTS: Thirty-seven male Fischer rats. INTERVENTIONS: Critical size defects 4 mm in diameter were created in the left mandibular bodies of the rats. The defects were filled with a bone marrow cell suspension (group 1), a synthetic bone matrix consisting of bovine collagen and calcium hydroxyapatite cement (group 2), the matrix and marrow cells (group 3), the matrix with 100 micro g of bone growth factor mixture (group 4), or the matrix with bone growth factor mixture and marrow cells (group 5). Animals were killed after 8 weeks, and the nondemineralized specimens were processed histologically. Specimens from group 1 were not processed because there was no grossly appreciable bone regeneration. Stereologic techniques were used to determine and compare the volume fractions and volume estimates of mature bone, new bone, osteoid, marrow, remaining cement, and fibrous tissue in each defect. RESULTS: Volumes of mature bone, new bone, and remaining cement did not differ significantly among the groups (P =.30 for mature bone, P =.17 for new bone, and P =.34 for cement). However, group 4 and 5 specimens contained significantly more osteoid and larger marrow spaces than did the group 2 and 3 specimens (P<.001 for both). The specimens in groups 2 and 3 contained significantly more fibrous tissue ingrowth than did those in groups 4 and 5 (P<.001). CONCLUSION: The synthetic bone substitute containing bone growth factor mixture was effective in stimulating new bone and osteoid development in the rat mandibular model.

Characterization of a developing lumbar arthrodesis in a sheep model with quantitative instability.

BACKGROUND CONTEXT: Mechanical forces have been considered responsible for stress shielding an arthrodesis, but the biology of a developing lumbar fusion has not been well characterized. PURPOSE: A large animal model was used to test the hypothesis that mechanical forces modify the biological processes involved in a developing bony fusion. STUDY DESIGN: Lumbar fusion was performed in an ovine model using custom instrumentation that permitted a controlled degree of anterior-posterior translation after surgery. Fusion sites were evaluated by radiography, microradiography, histology and histomorphometry at time points that corresponded with predicted early and later stages of bone healing. METHODS: Fourteen skeletally mature ewes underwent lumbar spinal fusion under general anesthesia. In the control (stable) group, the spine was rigidly fixed with a cage anteriorly and pedicle screws posteriorly. In the experimental (unstable) group, the spine was destabilized by an annulectomy (with no anterior implant) and custom pedicle screws that allowed 2 mm of anterior-posterior translation. Animals were euthanized 6 and 12 weeks after surgery. RESULTS: Radiographs confirmed that the fusion mass had not fully consolidated at either time point. Microradiographs revealed a trend toward increased bone formation at 6 weeks in the stable case as compared with the unstable, but by 12 weeks, this trend had reversed (p=.03). Intramembranous bone formation was the primary mechanism of healing near the transverse process in animals with both stable and unstable fixation. In the area between the two transverse processes, new bone formation occurred primarily through endochondral ossification. At 12 weeks, the stable case had significantly more cartilage formed (p=.023) but less newly formed bone (p=.07) as compared with the quantitatively unstable. CONCLUSIONS: This clinically realistic animal model allowed characterization of the biology of the developing arthrodesis before fusion. Under stable or unstable conditions, endochondral ossification was the predominant mechanism of new bone formation within the intertransverse process region. This finding, which contrasts with previous reports from small animal models of spine fusion, may reflect a difference in biology that results from the increased size of the intertransverse space in sheep as compared with small animals. Interestingly, mechanical instability increased the formation of new bone within this region, but not at the transverse process. Endochondral ossification therefore appears to respond to mechanical factors in the fusion site. The ovine model shows promise as an alternative to the rabbit model and may provide a more stringent test for potential new surgical and nonsurgical strategies for spine fusion.