自体成骨细胞3D生物复合打印颅骨骨瓣修复新西兰兔颅骨缺损模型的实验研究

Skull defect is a common neurosurgical disease. The result of cranioplasty and thepatient's neurological functional rehabilitation is influenced by the repair materials. 3D bio-printing of tissue engineering bone is a new choice in cranioplasty. In this study, 3D bio-printing of autologous osteoblasts was combined with lectrospinning intra-layer output to create multilayer printing and thus construct individualized simulated skulls. This new technique satisfies the mechanical requirements of skulls and maintains the cells activity. Moreover, the uniform cell distribution in the scaffold's 3D space, and the exchange of nutrients and metabolic waste can all be guaranteed. The simulated skulls were immediately transplanted to a New Zealand rabbit skull bone defect model. Then histopathologic, cell immunologic detections and imaging methods were used to observe the bone regeneration of osteoblasts, structural changes of scaffolds, and the healing of bone margins. The objective of this study is to find out a novel biomaterial for repair of skull bone defects and to build a new treatment mode for skull bone defect repair in clinic. More importantly, this study provides experimental basis and theoretical significances for 3D bio-printing of multi-material multicellular bone.

颅骨缺损是神经外科常见疾病，修补材料对颅骨修补手术是否成功及病人神经功能恢复和长期预后起着决定性作用。自体成骨细胞3D生物复合打印颅骨骨瓣是颅骨修补材料新的研究方向。本项目采用自体成骨细胞3D生物打印联合静电纺丝技术，同层输出、多层打印，构建仿真颅骨瓣，使打印的颅骨瓣精密个体化，达到颅骨物理学、生物学等要求。同时，使细胞在支架立体空间中均匀分布，维持细胞正常营养及代谢，保证细胞活性。将打印好的仿真颅骨瓣即时移植修复新西兰兔颅骨缺损模型。应用组织病理学、细胞免疫学及影像学等方法检测成骨细胞活性、支架结构变化、颅骨瓣与骨缘融合情况等。目的在于为颅骨缺损修补探索一种全新的生物材料，为临床颅骨缺损修补建立新的治疗模式，同时也为将来3D打印生物多材料、多细胞的骨组织工程器官提供重要的实验基础和理论依据。

颅骨缺损是神经外科常见疾病，修补材料对颅骨修补手术是否成功及病人神经功能恢复和长期预后起着决定性作用。自体成骨细胞3D生物复合打印颅骨骨瓣是颅骨修补材料新的研究方向。通过生长因子递送，共培养，机械刺激等促进3D生物支架中细胞的增殖，诱导分化。细胞在支架立体空间中均匀分布，维持细胞正常营养及代谢，保证细胞活性。与单独的PCL、PLGA、海藻酸钠、壳聚糖相比，3D生物支架可以模仿组织结构，与多种细胞结合，加快颅骨缺损的修补。另外，在支架中加入脐带内皮细胞可以促进血管的生成。支架的多孔结构可以保护细胞，同时支架中的细胞可以利用多孔结构将代谢废物排除和吸收营养物质。此外，将打印好的仿真颅骨瓣即时移植修复新西兰兔颅骨缺损模型。应用组织病理学、免疫组化及影像学等方法发现3D生物支架维持了成骨细胞活性、加进了颅骨缺损的修补，促进了颅骨瓣与骨缘融合。由此可见，3D生物打印支架在颅骨缺损修补方面具有潜在的应用价值。

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