Individual cell bioprinting to generate multi-tissue type condensations for osteochondral tissue regeneration

单个细胞生物打印可生成用于骨软骨组织再生的多组织类型浓缩物

• 批准号: 10659772
• 负责人: Eben Alsberg
• 金额: $ 40.36万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-05 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659772
• 关键词: 3-Dimensional; 3D Print; Address; Affect; Alginates; Architecture; Bathing; Biomimetics; Blood Vessels; Cartilage; Cell Communication; Cells; Chemicals; Clinical; Communities; Complex; Defect; Degenerative polyarthritis; Endothelial Cells; Engineering; Evaluation; Face; Fibrocartilages; Functional Regeneration; Gel; Geometry; Growth Factor; Health; Human; Hydrogels; Immune; Individual; Inferior; Inflammatory; Knee; Liquid substance; Location; Maintenance; Mechanics; Mesenchymal Stem Cells; Modeling; Modification; Movement; Operative Surgical Procedures; Orthopedics; Oryctolagus cuniculus; Patients; Persons; Phase; Phenotype; Physical condensation; Positioning Attribute; Printing; Procedures; Property; Reaction; Research Personnel; Resolution; Role; Shapes; Solid; Structure; Techniques; Technology; Therapeutic; Thick; Thinness; Tissue Engineering; Tissue constructs; Tissues; Vascularization; bioink; bioprinting; bioscaffold; bone; bone repair; cartilage repair; cartilaginous; design; endothelial stem cell; healing; human stem cells; improved; in vivo; innovation; novel; osteochondral tissue; osteogenic; particle; physical property; repaired; scaffold; spatiotemporal; standard of care; stem cells; three dimensional structure; tissue injury; tissue regeneration

Abstract Osteochondral defects of the knee are common worldwide, yet there are few viable options for patients with damaged osteochondral tissue as current treatments do not consistently regenerate functional tissue. The standard of care for osteochondral defect repair is arthroscopic microfracture surgery, but this procedure often results in formation of mechanically inferior fibrocartilage formation. To overcome limitations of this and other surgical procedures, tissue engineering strategies, such as cell-laden biomaterial scaffolds, are promising alternative approaches to treat these defects. However, scaffold-based strategies face several challenges, such as interference with critical cell-cell interactions, potential immune and/or inflammatory reaction to the scaffold and its degradation byproducts, and unsynchronized scaffold degradation rate with that of new tissue formation. New cellular condensation strategies without a scaffold address these issues, however, it is still difficult to precisely control the architecture of the engineered tissues to mimic the sophisticated three-dimensional (3D) structure and organization of natural osteochondral tissues and their structure-derived functions. Recently, 3D bioprinting has been applied in tissue engineering with the potential to create complicated, high-resolution 3D structures. In addition, we have engineered the first technology capable of 3D printing a cell-only bioink and maintaining the printed structure, which is necessary to form cell condensations. The hypothesis of this proposal is that cellular condensation-based prevascularized osteochondral tissue constructs of precisely defined geometries can be directly assembled with human stem cells and endothelial cells via 3D bioprinting into a photocurable liquid-like solid, shear-thinning and rapid self-healing microgel slurry with spatially controlled presentation of tissue specific growth factors. Microgel photocrosslinking after printing will provide temporary mechanical stability for the printed constructs during culture to permit cellular condensation formation. This cell- only bioprinting strategy will be implemented to print seamlessly continuous two-phase osteochondral tissue constructs with a prevascularized bone phase and a cartilage phase. Specifically, this proposal aims to (1) determine the role of microgel properties on the resolution and fidelity of the cell-only 3D printed constructs, (2) engineer prevascularized osteochondral constructs with individual cell-only bioinks by spatiotemporally controlled delivery of vasculogenic, osteogenic and chondrogenic growth factors, and (3) determine the clinical potential of the 3D printed prevascularized osteochondral constructs by evaluation of new osteochondral tissue formation and integration with the host vascular networks and bone and cartilage repair in a full-thickness osteochondral rabbit defect model. This platform strategy has the potential to greatly enhance the lives of those suffering from osteochondral defects and may enable the engineering of other complex functional tissues in the body.

抽象的 膝关节骨软骨缺损在世界范围内很常见，但对于患有膝关节骨软骨缺损的患者来说，几乎没有可行的选择 受损的骨软骨组织，因为目前的治疗方法不能持续再生功能组织。这 骨软骨缺损修复的护理标准是关节镜微骨折手术，但该手术通常 导致机械性能较差的纤维软骨形成。为了克服这个和其他的限制 外科手术、组织工程策略（例如载有细胞的生物材料支架）前景广阔 治疗这些缺陷的替代方法。然而，基于支架的策略面临着一些挑战，例如 干扰关键的细胞间相互作用、支架的潜在免疫和/或炎症反应 及其降解副产物，以及支架降解速率与新组织形成不同步。 没有支架的新细胞凝聚策略解决了这些问题，然而，仍然很难 精确控制工程组织的结构以模仿复杂的三维 (3D) 天然骨软骨组织的结构和组织及其结构衍生的功能。最近，3D 生物打印已应用于组织工程，具有创建复杂、高分辨率 3D 的潜力 结构。此外，我们还设计了第一项能够 3D 打印纯细胞生物墨水的技术， 保持印刷结构，这是形成细胞凝聚所必需的。本提案的假设 是基于细胞浓缩的预血管化骨软骨组织结构，其结构精确定义 几何形状可以通过 3D 生物打印直接与人类干细胞和内皮细胞组装成 空间控制的光固化类液固体、剪切稀化和快速自修复微凝胶浆料 组织特异性生长因子的呈现。打印后的微凝胶光交联将提供暂时的 培养过程中印刷结构的机械稳定性，以允许细胞凝结形成。这个细胞—— 将实施唯一的生物打印策略来打印无缝连续的两相骨软骨组织 具有预血管化骨相和软骨相的构造。具体而言，该提案旨在 (1) 确定微凝胶特性对纯细胞 3D 打印结构的分辨率和保真度的作用，(2) 通过时空设计，使用仅单个细胞的生物墨水来设计预血管化骨软骨结构 血管生长因子、成骨生长因子和软骨生长因子的受控递送，以及（3）确定临床 通过评估新骨软骨组织来评估 3D 打印预血管化骨软骨结构的潜力 与宿主血管网络的形成和整合以及全层骨和软骨的修复 兔骨软骨缺损模型。该平台战略有可能极大地改善人们的生活 患有骨软骨缺陷，并可能使工程中的其他复杂功能组织成为可能 身体。