High-resolution 3D printing of angle-ply annulus fibrosus scaffolds for intervertebral disc regeneration

Intervertebral disc (IVD) degeneration is one of the leading causes of disability, and current therapies are mainly unsatisfactory. The key pathological feature during IVD degeneration is the dysfunction of annulus fibrosus (AF). Although tissue-engineered AF has shown great promise for IVD regeneration, the design and fabrication of biomimetic AF scaffold remains a challenge due to the complexity of its structure. Nowadays, 3D printing technology has drawn great attention due to its customizable processes and ability to produce complex tissue architecture. However, few existing 3D printing methods can accurately replicate the fine angle-ply architecture of native AF, which is one of the most critical steps for IVD regeneration, due to the limited printing resolution. In this study, we aimed to fabricate high-resolution polycaprolactone (PCL) scaffolds using a newly developed electrohydrodynamic 3D printing technique. The structural advantages of such scaffolds were verified by finite element analysis (FEA). The PCL scaffolds were further assembled into AF construct to replicate the angle-ply architecture of AF. The optimal assembling method was confirmed by FEA and mechanical tests. Thein vitroexperiments showed that the 3D printed AF scaffolds presented favorable biocompatibility and supported the adhesion and growth of AF cells. Thein vivoperformance of tissue-engineered IVDs (TE-IVDs), which consisted of 3D printed AF scaffold and GelMA hydrogel that simulated nucleus pulposus (NP), were evaluated using a rat total disc replacement model. We found that the implantation of TE-IVDs helped maintain the disc height, reduced the loss of NP water content, and partially restored the biomechanical function of IVD. In addition, the TE-IVDs achieved well integration with adjacent tissues and promoted new tissue formation. In summary, being able to accurately simulate the structural characteristics of native AF, the 3D printed angle-ply AF scaffolds hold potential for future applications in IVD regeneration.