Multi-tissue type condensations for trachea tissue regeneration via individual cell bioprinting

通过单细胞生物打印进行气管组织再生的多组织类型浓缩

• 批准号: 10643041
• 负责人: Eben Alsberg
• 金额: --
• 依托单位: JESSE BROWN VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10643041
• 关键词: 3-Dimensional; 3D Print; Ablation; Address; Affect; Anastomosis - action; Animals; Autologous; Bacterial Infections; Bathing; Benign; Biochemical; Biocompatible Materials; Blood Vessels; Cartilage; Cell Communication; Cell Density; Cell Proliferation; Cell physiology; Cells; Cessation of life; Chondrogenesis; Complex; Defect; Devices; Engineering; Epithelial Cells; Epithelium; Extracellular Matrix; Gel; Gelatin; Geometry; Growth Factor; Gunshot wound; Histologic; Histology; Human; Immune response; Implant; Individual; Inflammatory Response; Intubation; Lasers; Legal patent; Life; Maintenance; Malignant Neoplasms; Manuals; Mechanics; Mesenchymal Stem Cells; Military Personnel; Modeling; Mucous Membrane; Nutrient; Organ; Oryctolagus cuniculus; Pain; Patients; Penetration; Performance; Phenotype; Physical condensation; Physiological; Polymers; Printing; Production; Quality of life; Radial; Resolution; Respiratory Failure; Role; Soldier; Source; Stents; Structure; Technology; Testing; Therapeutic; Tissue Differentiation; Tissue Engineering; Tissue constructs; Tissues; Trachea; Tracheal Stenosis; Tracheostomy procedure; Trauma; Tube; Tubular formation; Vascular System; Vascularization; Veterans; Work; airway epithelium; bioink; biomechanical test; bioprinting; bioscaffold; bronchial epithelium; cartilaginous; cell type; clinically relevant; combat; crosslink; design; disability; effective therapy; endothelial stem cell; flexibility; healing; in vivo; manufacture; mechanical properties; military veteran; particle; physical property; prevent; respiratory; restenosis; scaffold; self assembly; tissue regeneration

Abstract As a result of prolonged intubation, tracheostomy, external trauma, penetrating fragment projectiles, gunshot wounds and improvised explosive devices during combat, and benign or malignant tumors, many soldiers and veterans in the US military suffer from severe trachea stenosis or damage that can cause complete airway failure, Since there is no successful long-term treatment for long-segment tracheal stenosis or damage, tissue engineering strategies have been explored to develop neotracheas using different combinations of biomaterials and cell sources. However, biomaterial scaffold-based approaches often interfere with critical cell-cell interactions, cell proliferation and new extracellular matrix production that are important during the formation of functional trachea tissue. A functional replacement trachea must retain (1) radial rigidity to prevent restenosis, (2) anastomose with host vasculature to adequately provide nutrients to the implant, and (3) contain respiratory epithelium to provide a protective mucosal layer. Combining three-dimensional (3D) bioprinting technologies with scaffold-free tissue engineering principles presents a powerful platform for engineering a multi-tissue functional trachea, and would circumvent the aforementioned limitations of scaffold-based approaches. This proposal aims to leverage the benefits of our recently developed individual cell-only 3D bioprinting technology, which allows for printing of complex and high-resolution cell condensation-based tissue constructs to engineer functional tracheas. We plan to print scaffold-free, multi-tissue neotracheas using multiple discrete individual cell-only bioinks for spatially distinct differentiation of tissue types driven by spatially controlled presentation of tissue- specific growth factors. Construct self-assembly will be driven by the condensation of autologously sourced human mesenchymal stem cells (hMSCs) for cartilaginous tissue and autologous endothelial progenitor cells and hMSCs for prevascular tissue, with autologous human bronchial epithelial cells applied to line the lumen of the neotrachea. Specifically, this proposal aims to (1) examine the role of the physical properties of the microgel support slurry on cell-only bioink printing, condensation formation/maintenance, and chondrogenesis of the 3D bioprinted structures, (2) 3D bioprint cartilage ring constructs with chondrogenic bioink and prevascularized ring constructs with vasculogenic bioink using the individual cell-only bioprinting technology, and (3) engineer prevascularized and epithelized tracheal tissue with chondrogenic and vasculogenic bioinks using the individual cell-only bioprinting technology. As an exploratory aim, the capacity of the engineered tracheas to restore airway functionality will be evaluated in an animal defect model. This work ultimately seeks to utilize a facile and flexible individual cell-only bioink 3D printing platform to engineer a patient-specific replacement trachea that provides requisite physiologic and mechanical properties for replacement in those that are affected by long-segment tracheal stenosis. The inherent flexibility of this individual cell-only 3D printing platform to create complex structures composed of multiple spatially distinct tissue types can be leveraged to develop other multi-tissue organs.

抽象的 由于长时间插管、气管切开、外伤、穿透性碎片射弹、枪伤 战斗中的伤口和简易爆炸装置，以及良性或恶性肿瘤，许多士兵和 美国退伍军人患有严重的气管狭窄或损伤，可能导致气道完全衰竭， 由于长段气管狭窄或损伤尚无成功的长期治疗方法，组织 已经探索了使用不同的生物材料组合来开发新气管的工程策略 和细胞来源。然而，基于生物材料支架的方法通常会干扰关键的细胞-细胞 相互作用、细胞增殖和新的细胞外基质产生，这些在细胞形成过程中非常重要 功能性气管组织。功能性替换气管必须保持 (1) 径向刚性以防止再狭窄， (2) 与宿主脉管系统吻合，为植入物提供充足的营养，以及 (3) 包含呼吸系统 上皮细胞提供保护性粘膜层。将三维 (3D) 生物打印技术与 无支架组织工程原理为工程多组织功能提供了一个强大的平台 气管，并且将规避基于支架的方法的上述限制。该提案旨在 利用我们最近开发的单细胞 3D 生物打印技术的优势，该技术允许 打印复杂且高分辨率的基于细胞浓缩的组织结构以设计功能 气管。我们计划使用多个离散的单个细胞打印无支架的多组织新气管 生物墨水，用于由组织的空间控制呈现驱动的组织类型的空间明显分化 特定的生长因子。构建自组装将由自体来源的凝结驱动 用于软骨组织和自体内皮祖细胞的人间充质干细胞 (hMSC) 和hMSCs用于血管前组织，自体人支气管上皮细胞应用于排列血管内腔 新气管。具体来说，该提案旨在（1）检查微凝胶物理性质的作用 支持纯细胞生物墨水打印、凝结形成/维护和 3D 软骨形成的浆料 生物打印结构，(2) 具有软骨形成生物墨水和预血管化环的 3D 生物打印软骨环结构 使用仅单个细胞的生物打印技术构建血管生成生物墨水，以及 (3) 工程师 使用个体使用软骨形成和血管形成生物墨水来预血管化和上皮化气管组织 仅细胞生物打印技术。作为一个探索性目标，工程气管恢复气道的能力 将在动物缺陷模型中评估功能。这项工作最终寻求利用一种简便灵活的方法 仅单个细胞的生物墨水 3D 打印平台，用于设计患者特定的替换气管，提供 受长节段影响的患者需要更换所需的生理和机械特性 气管狭窄。这种单独的纯细胞 3D 打印平台具有固有的灵活性，可以创建复杂的 由多种空间上不同的组织类型组成的结构可用于开发其他多组织 器官。