A Rab33b missense mouse model for Smith-McCort dysplasia shows bone resorption defects and altered protein glycosylation.

Smith McCort (SMC) dysplasia is a rare, autosomal recessive, osteochondrodysplasia that can be caused by pathogenic variants in either RAB33B or DYM genes. These genes codes for proteins that are located at the Golgi apparatus and have a role in intracellular vesicle trafficking. We generated mice that carry a Rab33b disease-causing variant, c.136A>C (p.Lys46Gln), which is identical to that of members from a consanguineous family diagnosed with SMC. In male mice at 4 months of age, the Rab33b variant caused a mild increase in trabecular bone thickness in the spine and femur and in femoral mid-shaft cortical thickness with a concomitant reduction of the femoral medullary area, suggesting a bone resorption defect. In spite of the increase in trabecular and cortical thickness, bone histomorphometry showed a 4-fold increase in osteoclast parameters in homozygous Rab33b mice suggesting a putative impairment in osteoclast function, while dynamic parameters of bone formation were similar in mutant versus control mice. Femur biomechanical tests showed an increased in yield load and a progressive elevation, from WT to heterozygote to homozygous mutants, of bone intrinsic properties. These findings suggest an overall impact on bone material properties which may be caused by disturbed protein glycosylation in cells contributing to skeletal formation, supported by the altered and variable pattern of lectin staining in murine and human tissue cultured cells and in liver and bone murine tissues. The mouse model only reproduced some of the features of the human disease and was sex-specific, manifesting in male but not female mice. Our data reveal a potential novel role of RAB33B in osteoclast function and protein glycosylation and their dysregulation in SMC and lay the foundation for future studies.

Discovery of small molecule agonists of the Relaxin Family Peptide Receptor 2.

The relaxin/insulin-like family peptide receptor 2 (RXFP2) belongs to the family of class A G-protein coupled receptors (GPCRs) and it is the only known target for the insulin-like factor 3 peptide (INSL3). The importance of this ligand-receptor pair in the development of the gubernacular ligament during the transabdominal phase of testicular descent is well established. More recently, RXFP2 has been implicated in maintaining healthy bone formation. In this report, we describe the discovery of a small molecule series of RXFP2 agonists. These compounds are highly potent, efficacious, and selective RXFP2 allosteric agonists that induce gubernacular invagination in mouse embryos, increase mineralization activity in human osteoblasts in vitro, and improve bone trabecular parameters in adult mice. The described RXFP2 agonists are orally bioavailable and display favorable pharmacokinetic properties, which allow for future evaluation of the therapeutic benefits of modulating RXFP2 activation in disease models.

Mechanical Analysis of 3 Posterior Fusion Assemblies Intended to Cross the Cervicothoracic Junction.

STUDY DESIGN: This was a biomechanical comparison study. OBJECTIVE: The objective of this study is to evaluate the mechanical properties of 3 posterior spinal fusion assemblies commonly used to cross the cervicothoracic junction. SUMMARY OF BACKGROUND: When posterior cervical fusions are extended into the thoracic spine, an instrumentation transition is often utilized. The cervical rod (3.5 mm) can continue using thoracic screws designed to accept the cervical rods. Alternatively, traditional thoracic screws may be used to accept thoracic rods (5.5 mm). This requires the use of a 3.5-5.5 mm transition rod or a separate 5.5 mm rod and a connector to fix the 3.5 and 5.5 mm rod together. Fusion success depends on the immobilization of vertebrae, yet the mechanics provided by these different assemblies are unknown. MATERIALS AND METHODS: Three titanium alloy posterior fusion assemblies intended to cross the cervicothoracic junction underwent static compressive bending, tensile bending, and torsion as described in ASTM F1717 to a torque of 2.5 Nm. Five samples of each assembly were attached to ultrahigh molecular weight polyethylene blocks via multiaxial screws for testing. Force and displacement were recorded, and the stiffness of each construct was calculated. RESULTS: The 2 assemblies that included a 5.5 mm rod were found to be stiffer and have less range of motion than the assembly that used only 3.5 mm rods. CONCLUSIONS: The results of this study indicate that incorporating a 5.5 mm rod in a fusion assembly adds significant stiffness to the construct. When the stability of a fusion is of heightened concern, as demonstrated by the ASTM F1717 vertebrectomy (worst-case scenario) model, including 5.5 mm rods may increase fusion success rates. LEVEL OF EVIDENCE: Level V.

Augmentation of core decompression with synthetic bone graft does not improve mechanical properties of the proximal femur.

Core decompression is a minimally invasive surgical technique used to treat patients with avascular necrosis of the femoral head. The procedure requires an entry hole in the lateral cortex of the femur which potentially leaves patients susceptible to subtrochanteric fractures. The purpose of this study was to determine if filling the core decompression tract with synthetic bone-graft mechanically strengthens the proximal femur. Twenty composite synthetic femurs underwent a core decompression procedure; ten were augmented with synthetic bone-graft (PRO-DENSE™, Wright Medical) and ten femurs were left unfilled as a control group. Compressive testing to failure was performed using a mechanical testing machine. Stiffness, fracture load, and toughness did not significantly differ between groups. More subtrochanteric fractures were seen in the control group (6 of 10 specimens) compared to the bone-graft augmented group (2 of 10 specimens). In conclusion, augmentation of a core decompression tract does not improve mechanical properties in a synthetic bone model but may be protective of subtrochanteric fracture.

Validation of a custom spine biomechanics simulator: A case for standardization.

Mechanical testing machines used in cadaveric spine biomechanics research vary between labs. It is a necessary first step to understand the capabilities and limitations in any testing machine prior to publishing experimental data. In this study, a reproducible protocol that uses a synthetic spine was developed and used to quantify the inherent rotation error and the ability to apply loads in a single physiologic plane (pure-moment) of a custom spine biomechanics simulator. Rotation error was evaluated by comparing data collected by the test machine and the data collected by an optical motion capture system. Pure-moment loading was assessed by comparing the out-of-plane loads to the primary plane load. Using synthetic functional spine units previously shown to have mechanics similar to the cadaveric human spine, the simulator was evaluated using a dynamic test protocol reflective of its future use in the study of cadaveric spine specimens. Rotation errors inherent in the test machine were <0.25° compared to motion capture. Out of plane loads were <4.0% of the primary plane load, which confirmed pure-moment loading. The authors suggest that a standard validation protocol for biomechanical spine testing machines is needed for transparency and accurate field-wide data interpretation and comparison. We offer recommendations based on the reproducible use of a synthetic spinal specimen for consideration.