Kérourédan Olivia, Washio Ayako, Handschin Charles, Devillard Raphaël, Kokabu Shoichiro, Kitamura Chiaki, Tabata Yasuhiko
INSERM, U1026 BIOTIS, University of Bordeaux, 146 rue Léo Saignat, Bordeaux 33076, France.
Faculty of Dentistry, University of Bordeaux, 146 rue Léo Saignat, Bordeaux 33076, France.
Biomed Mater. 2024 Feb 16;19(2). doi: 10.1088/1748-605X/ad270a.
Despite significant advances in the management of patients with oral cancer, maxillofacial reconstruction after ablative surgery remains a clinical challenge. In bone tissue engineering, biofabrication strategies have been proposed as promising alternatives to solve issues associated with current therapies and to produce bone substitutes that mimic both the structure and function of native bone. Among them, laser-assisted bioprinting (LAB) has emerged as a relevant biofabrication method to print living cells and biomaterials with micrometric resolution onto a receiving substrate, also called 'biopaper'. Recent studies have demonstrated the benefits of prevascularization using LAB to promote vascularization and bone regeneration, but mechanical and biological optimization of the biopaper are needed. The aim of this study was to apply gelatin-sheet fabrication process to the development of a novel biopaper able to support prevascularization organized by LAB for bone tissue engineering applications. Gelatin-based sheets incorporating bioactive glasses (BGs) were produced using various freezing methods and crosslinking (CL) parameters. The different formulations were characterized in terms of microstructural, physical, mechanical, and biological properties in monoculture and coculture. Based on multi-criteria analysis, a rank scoring method was used to identify the most relevant formulations. The selected biopaper underwent additional characterization regarding its ability to support mineralization and vasculogenesis, its bioactivity potential anddegradability. The biopaper 'Gel5wt% BG1wt%-slow freezing-CL160 °C 24 h' was selected as the best candidate, due to its suitable properties including high porosity (91.69 ± 1.55%), swelling ratio (91.61 ± 0.60%), Young modulus (3.97 × 10± 0.97 × 10Pa) but also its great cytocompatibility, osteogenesis and bioactivity properties. The preorganization of human umbilical vein endothelial cell using LAB onto this new biopaper led to the formation of microvascular networks. This biopaper was also shown to be compatible with 3D-molding and 3D-stacking strategies. This work allowed the development of a novel biopaper adapted to LAB with great potential for vascularized bone biofabrication.
尽管口腔癌患者的治疗取得了显著进展，但切除术后的颌面重建仍然是一项临床挑战。在骨组织工程中，生物制造策略已被提出作为有前景的替代方法，以解决当前治疗相关的问题，并生产出模仿天然骨结构和功能的骨替代物。其中，激光辅助生物打印（LAB）已成为一种相关的生物制造方法，可将活细胞和生物材料以微米级分辨率打印到接收基板上，该基板也称为“生物纸”。最近的研究表明，使用LAB进行预血管化有助于促进血管化和骨再生，但生物纸的机械和生物学性能仍需优化。本研究的目的是将明胶片制造工艺应用于新型生物纸的开发，该生物纸能够支持LAB组织的预血管化，用于骨组织工程应用。采用不同的冷冻方法和交联（CL）参数制备了含有生物活性玻璃（BGs）的明胶基片。对不同配方在单培养和共培养中的微观结构、物理、机械和生物学性能进行了表征。基于多标准分析，采用等级评分法确定最相关的配方。对所选生物纸在支持矿化和血管生成的能力、生物活性潜力和降解性方面进行了额外的表征。生物纸“Gel5wt% BG1wt%-缓慢冷冻-CL160 °C 24 h”被选为最佳候选者，因为它具有合适的性能，包括高孔隙率（91.69 ± 1.55%）、溶胀率（91.61 ± 0.60%）、杨氏模量（3.97 × 10± 0.97 × 10Pa），以及良好的细胞相容性、成骨和生物活性性能。使用LAB将人脐静脉内皮细胞预组织到这种新型生物纸上，导致形成微血管网络。该生物纸还被证明与3D成型和3D堆叠策略兼容。这项工作使得开发出一种适用于LAB的新型生物纸，具有用于血管化骨生物制造的巨大潜力。
