Vascularization is critical for skull development, maintenance, and healing. Yet, there remains a significant knowledge gap in the relationship of blood vessels to cranial skeletal progenitors during these processes. Here, we introduce a quantitative 3D imaging platform to enable the visualization and analysis of high-resolution data sets (>100 GB) throughout the entire murine calvarium. Using this technique, we provide single-cell resolution 3D maps of vessel phenotypes and skeletal progenitors in the frontoparietal cranial bones. Through these high-resolution data sets, we demonstrate that CD31^hiEmcn^hi vessels are spatially correlated with both Osterix+ and Gli1+ skeletal progenitors during postnatal growth, healing, and stimulated remodeling, and are concentrated at transcortical canals and osteogenic fronts. Interestingly, we find that this relationship is weakened in mice with a conditional knockout of PDGF-BB in TRAP+ osteoclasts, suggesting a potential role for osteoclasts in maintaining the native cranial microvascular environment. Our findings provide a foundational framework for understanding how blood vessels and skeletal progenitors spatially interact in cranial bone, and will enable more targeted studies into the mechanisms of skull disease pathologies and treatments. Additionally, our technique can be readily adapted to study numerous cell types and investigate other elusive phenomena in cranial bone biology.

Low oxygen (O₂) diffusion into large tissue engineered scaffolds hinders the therapeutic efficacy of transplanted cells. To overcome this, we previously studied hollow, hyperbarically-loaded microtanks (μtanks) to serve as O₂ reservoirs. To adapt these for bone regeneration, we fabricated biodegradable μtanks from polyvinyl alcohol and poly(lactic-co-glycolic acid) and embedded them to form 3D-printed, porous poly-ε-caprolactone (PCL)-μtank scaffolds. PCL-μtank scaffolds were loaded with pure O₂ at 300 – 500 psi. When placed at atmospheric pressures, the scaffolds released O₂ over a period of up to 8 hours. We confirmed the inhibitory effects of hypoxia on the osteogenic differentiation of human adipose-derived stem cells (hASCs and we validated that μtank-mediated transient hyperoxia had no toxic impacts on hASCs, possibly due to upregulation of endogenous antioxidant regulator genes. We assessed bone regeneration in vivo by implanting O₂-loaded, hASC-seeded, PCL-μtank scaffolds into murine calvarial defects (4 mm diameters × 0.6 mm height) and subcutaneously (4 mm diameter × 8 mm height). In both cases we observed increased deposition of extracellular matrix in the O₂ delivery group along with greater osteopontin coverages and higher mineral deposition. This study provides evidence that even short-term O₂ delivery from PCL-μtank scaffolds may enhance hASC-mediated bone tissue regeneration.

An algorithm for automatic spinal pedicle screw planning is reported and evaluated in simulation and first clinical studies. A statistical atlas of the lumbar spine ( N = 40 members) was constructed for active shape model (ASM) registration of target vertebrae to an unsegmented patient CT. The atlas was augmented to include ‘reference’ trajectories through the pedicles as defined by a spinal neurosurgeon. Following ASM registration, the trajectories are transformed to the patient CT and accumulated to define a patient-specific screw trajectory, diameter, and length. The algorithm was evaluated in leave-one-out analysis ( N = 40 members) and for the first time in a clinical study ( N = 5 patients undergoing cone-beam CT (CBCT) guided spine surgery), and in simulated low-dose CBCT images. ASM registration achieved (2.0 ± 0.5) mm root-mean-square-error (RMSE) in surface registration in 96% of cases, with outliers owing to limitations in CT image quality (high noise/slice thickness). Trajectory centerlines were conformant to the pedicle in 95% of cases. For all non-breaching trajectories, automatically defined screw diameter and length were similarly conformant to the pedicle and vertebral body (98.7%, Grade A/B). The algorithm performed similarly in CBCT clinical studies (93% centerline and screw conformance) and was consistent at the lowest dose levels tested. Average runtime in planning five-level (lumbar) bilateral screws (ten trajectories) was (312.1 ± 104.0) s. The runtime per level for ASM registration was (41.2 ± 39.9) s, and the runtime per trajectory was (4.1 ± 0.8) s, suggesting a runtime of ~(45.3 ± 39.9) s with a more fully parallelized implementation. The algorithm demonstrated accurate, automatic definition of pedicle screw trajectories, diameter, and length in CT images of the spine without segmentation. The studies support translation to clinical studies in free-hand or robot-assisted spine surgery, quality assurance, and data analytics in which fast trajectory definition is a benefit to workflow.

Porous Magnesium (Mg) is a promising biodegradable scaffold for treating critical-size bone defects, and as an essential element for human metabolism, Mg has shown sufficient biocompatibility. Its elastic moduli and yield strengths are closer to those of cortical bone than common, inert metallic implants, effectively reducing stress concentrations around host tissue as well as stress shielding. More importantly, Mg can degrade and be absorbed in the human body in a safe and controlled manner, thereby reducing the need for second surgeries to remove implants. The development of porous Mg scaffolds via conventional selective laser melting techniques has been limited due to Mg’s low boiling point, high vapor pressures, high reactivity, and non-ideal microstructures in additively manufactured parts. Here we present an exciting alternative to conventional additive techniques: 3D weaving with Mg wires that have controlled chemistries and microstructures. The weaving process offers high throughput manufacturing as well as porous architectures that can be optimized for stiffness and porosity with topology optimization. Once woven, we dip-coat the weaves with polylactic acid to enhance their strength and corrosion resistance. Following fabrication, we characterize their mechanical properties, corrosion behavior, and cell compatibility in vitro, and we use an intramuscular implantation model to evaluate their in vivo corrosion behavior and tissue response.