Bioprinted anisotropic scaffolds with fast stress relaxation bioink for engineering 3D skeletal muscle and repairing volumetric muscle loss.

Viscoelastic hydrogels can enhance 3D cell migration and proliferation due to the faster stress relaxation promoting the arrangement of the cellular microenvironment. However, most synthetic photocurable hydrogels used as bioink materials for 3D bioprinting are typically elastic. Developing a photocurable hydrogel bioink with fast stress relaxation would be beneficial for 3D bioprinting engineered 3D skeletal muscles in vitro and repairing volumetric muscle loss (VML) in vivo; however, this remains an ongoing challenge. This study aims to develop an interpenetrating network (IPN) hydrogel with tunable stress relaxation using a combination of gelatin methacryloyl (GelMA) and fibrinogen. These IPN hydrogels with faster stress relaxation showed higher 3D cellular proliferation and better differentiation. A 3D anisotropic biomimetic scaffold was further developed via a printing gel-in-gel strategy, where the extrusion printing of cell-laden viscoelastic FG hydrogel within Carbopol supported gel. The 3D engineered skeletal muscle tissue was further developed via 3D aligned myotube formation and contraction. Furthermore, the cell-free 3D printed scaffold was implanted into a rat VML model, and both the short and long-term repair results demonstrated its ability to enhance functional skeletal muscle tissue regeneration. These data suggest that such viscoelastic hydrogel provided a suitable 3D microenvironment for enhancing 3D myogenic differentiation, and the 3D bioprinted anisotropic structure provided a 3D macroenvironment for myotube organization, which indicated the potential in skeletal muscle engineering and VML regeneration. STATEMENT OF SIGNIFICANCE: The development of a viscoelastic 3D aligned biomimetic skeletal muscle scaffold has been focused on skeletal muscle regeneration. However, a credible technique combining viscoelastic hydrogel and printing gel-in-gel strategy for fabricating skeletal muscle tissue was rarely reported. Therefore, in this study, we present an interpenetrating network (IPN) hydrogel with fast stress relaxation for 3D bioprinting engineered skeletal muscle via a printing gel-in-gel strategy. Such IPN hydrogels with tunable fast stress relaxation resulted in high 3D cellular proliferation and adequate differentiation in vitro. Besides, the 3D hydrogel-based scaffolds also enhance functional skeletal muscle regeneration in situ. We believe that this study provides several notable advances in tissue engineering that can be potentially used for skeletal muscle injury treatment in clinical.

Cervical Sagittal Alignment in Patients With Basilar Invagination.

STUDY DESIGN: Retrospective study. OBJECTIVE: To present a morphological map of cervical sagittal alignment in basilar invagination (BI), a congenital anomaly of the craniovertebral junction, and contribute to a comprehensive understanding of cervical sagittal alignment in congenital cervical deformities. SUMMARY OF BACKGROUND DATA: Ideal cervical sagittal alignment and surgical targets are debated by scholars. However, most of the literature focuses on the description of cervical sagittal alignment in acquired cervical diseases and normal subjects and few on congenital cervical spine deformities. MATERIALS AND METHODS: This study analyzed cervical spine lateral radiographs of 87 BI patients and 98 asymptomatic subjects. They were analyzed for cranial, cervical spine, and thoracic inlet parameters. RESULTS: Patients with BI manifested significantly larger values for the following parameters than asymptomatic subjects: cranial tilt, cranial incidence angle, sagittal vertical axis (SVA) CGH-C7, C2-C7 angle, cervical tilt, and significantly smaller values for the following parameters: cranial slope, C0-C2 angle, C0-C7 angle, SVA C2-C7, spine tilt, thoracic inlet angle, and neck tilt. In the BI group, SVA C2-C7 was the cervical parameter most strongly correlated with the cranial, cervical spine, and thoracic inlet parameters, and was smaller in BI patients with fusion (atlanto-occipital assimilation) than in those without. CONCLUSION: A significant difference was observed between BI patients and asymptomatic subjects. BI patients have craniums tilted forward and downward, smaller upper cervical lordosis, larger lower cervical lordosis, and smaller thoracic inlet angle. In BI patients, the SVA C2-C7 is an important parameter in cervical sagittal alignment. In both individuals with congenital anomalies of the craniovertebral junction and the asymptomatic population, cervical spine alignment is significantly associated with cranial alignment, particularly thoracic inlet alignment.

Injectable remote magnetic nanofiber/hydrogel multiscale scaffold for functional anisotropic skeletal muscle regeneration.

Developing an injectable anisotropic scaffold with precisely topographic cues to induce 3D cellular organization plays a critical role in volumetric muscle loss (VML) repair in vivo. However, controlling aligned myofiber regeneration in vivo based on previous injectable scaffolds continues to prove challenging, especially in a 3D configuration. Herein, we prepare the monodisperse remote magnetic controlled short nanofibers (MSNFs) with a high yield using an advanced coaxial electrospinning-cyrocutting method. An injectable anisotropic MSNF/Gel nanofiber/hydrogel scaffold based on MSNFs within photocurable hydrogel is further designed, showing the ability to guide 3D cellular alignment and organization by the precise microarchitecture control via a remote magnetic field. MSNF/Gel anisotropic scaffolds were able to recreate the macroscale and microscale topographical features of orbicular muscle and bipennate muscle mimicking their anatomical locations. Furthermore, the resultant MSNF/Gel anisotropic scaffolds significantly enhanced aligned myofiber formation in vivo and improved functional recovery of injured muscles in animal VML models. In summary, this approach offers a new promising tissue engineering strategy not only for the aligned myofiber formation for enhancing skeletal muscle regeneration in vivo but also for other biofabrication of living constructs containing complex anisotropy in vitro.

Anatomical analysis of the C2 pedicle in patients with basilar invagination.

PURPOSE: To evaluate and describe the morphologic features of the C2 pedicle in patients with basilar invagination (BI) for informing the placement of pedicle screws. C2 pedicle screw placement is an important surgical technique for the treatment of atlantoaxial instability in patients with BI. However, no systematic and comprehensive anatomical study of the C2 pedicle in patients with BI has been reported. METHODS: The data from 100 patients diagnosed with BI (BI group) and 100 patients without head or cervical disease (control group) were included in the study. Radiographic parameters, including the pedicle width, length, height, transverse angle, lamina angle, and superior angle, were measured and analyzed on CT images. After summary analysis, the effect of C2-3 congenital fusion on C2 pedicle deformity in patients with BI was also investigated. RESULTS: The width, length, and height of the C2 pedicle of the BI patients were smaller than those of the control group. The pedicle cancellous bone was smaller in the BI group, while no significant difference in cortical bone was observed. In total, 44% of the pedicles were smaller than 4.5 mm in the BI group. Patients with C2-3 congenital fusion presented with smaller pedicle transverse angles and larger pedicle superior angles than those without fusion. Wide variations in the left and right angles of the pedicle were observed in the BI group with atlantoaxial dislocation or atlantooccipital fusion. CONCLUSION: The C2 pedicle in the BI group was thinner than that in the control group due to a smaller cortical bone. Cases of C2-3 congenital fusion, atlantoaxial dislocation, and atlantooccipital fusion displayed variation in the angle of the C2 pedicle.

3D bioprinted multiscale composite scaffolds based on gelatin methacryloyl (GelMA)/chitosan microspheres as a modular bioink for enhancing 3D neurite outgrowth and elongation.

The integration of multiscale micro- and macroenvironment has been demonstrated as a critical role in designing biomimetic scaffolds for peripheral nerve tissue regeneration. While it remains a remarkable challenge for developing a biomimetic multiscale scaffold for enhancing 3D neuronal maturation and outgrowth. Herein, we present a 3D bioprinted multiscale scaffold based on a modular bioink for integrating the 3D micro- and macroenvironment of native nerve tissue. Gelatin methacryloyl (GelMA)/Chitosan Microspheres (GC-MSs) were prepared by a microfluidic approach, and the effect of these microspheres on enhancing neurite outgrowth and elongation of PC12 cells was demonstrated. The 3D multiscale composite scaffolds were bioprinted based on microspheres and hydrogel as the modular bioink. The co-culture of PC12 cells and RSC96 Schwann cells within these 3D biomimetic scaffolds were investigated to evaluate such a 3D multiscale environment for neurite outgrowth and Schwann cell proliferation. These results indicate that such multiscale composite scaffold with hydrogel microspheres provided a suitable 3D microenvironment for enhancing neurite growth, and the 3D printed hydrogel network provided a 3D macroenvironment mimicking the epineurium layer for Schwann cells proliferation and nerve cell organization, which is promising for the great potential applications in nerve tissue engineering.