Development of a multifunctional, acoustofluidic 3D bioprinter with single-cell resolution

开发具有单细胞分辨率的多功能声流控 3D 生物打印机

• 批准号: 10596518
• 负责人: Zhenhua Tian
• 金额: $ 34.98万
• 依托单位: VIRGINIA POLYTECHNIC INST AND ST UNIV
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10596518
• 关键词: 3-Dimensional; 3D Print; Acoustics; Address; Anisotropy; Architecture; Area; Biological; Biological Models; Biomedical Research; Biomimetics; Blood Vessels; Blood capillaries; Cell Communication; Cell Density; Cell Shape; Cell physiology; Cells; Communities; Complex; Development; Diameter; Drug Screening; Emerging Technologies; Ensure; Face; Holography; Hydrogels; Implant; In Vitro; Individual; Ink; Light; Microfluidics; Mississippi; Motor Neurons; Muscle; Muscle Fibers; Pattern; Performance; Physiological; Polymers; Positioning Attribute; Printing; Property; Protocols documentation; Regenerative Medicine; Research; Research Personnel; Resolution; Rotation; Series; Skeletal Muscle; Structure; Surface; System; Technology; Tissue Engineering; Tissue Model; Tissues; Vascular System; Vascularization; anticancer research; biomaterial compatibility; bioprinting; cell type; density; digital; improved; in vitro Model; in vivo; instrument; internal control; invention; large print; meter; microsystems; neuromuscular; next generation; photopolymerization; polymerization; prototype; scaffold; tool; tumor

Project Summary Three-dimensional (3D) bioprinting is a rapidly emerging technology that has the potential to quickly print customized, functional, biological tissues. In recent years, much progress has been made in modifying traditional printing systems for 3D bioprinting. However, their printed tissues often lack the resolution and complexity necessary to achieve the essential functions, physiological conditions, and anisotropic properties found in native tissues. Therefore, there is a critical need to develop next-generation 3D bioprinting instruments that address the following three key technologic limitations: (1) the inability to control internal cell positions in printed matrix material voxels and achieve the desired cell-cell spacing (i.e., cell proximity resolution < 10 µm) that is critical to ensure proper tissue functions from the cellular level and studying cell-cell interactions in the 3D microenvironments; (2) the inability to print tissues with high local cell densities (>109 cells/mL) as observed in vivo; and (3) the inability to perform scaffold-free printing of large-scale tissues with multiscale biomimetic cellular architectures (e.g., cell pattern and alignment), which are essential to achieve desired anisotropic tissue properties and key functions that depend on multiscale cell arrangements. Over the last ten years, we have developed a series of acoustofluidic (i.e., the fusion of acoustic and microfluidic) technologies, which are excellent candidates to address the bottlenecks above. In particular, we have recently developed acoustofluidic holography, an acoustics-based, biocompatible, and high-resolution cell manipulation technology that allows one to pattern, rotate, and concentrate cell seeded matrix materials before polymerization. Building upon this technology, in this R01 project, we propose to develop and validate an acoustofluidic 3D bioprinting prototype to print functional tissues with high cell proximity resolution (<10 µm) and complex features (such as biomimetic cellular architectures, controlled anisotropic properties, and high cell densities) in a biocompatible, fast, and scalable manner. Our acoustofluidic 3D bioprinter will be validated by printing vascularized tumor spheroids with stroma and anisotropic, innervated, vascularized skeletal muscle tissues. Compared to current 3D bioprinting instruments, our acoustofluidic 3D bioprinter will have multiple advantages including: (1) ability to control internal cell positions of printed matrix voxels and achieve high cell proximity resolution (< 10 µm); (2) ability to print tissues with high local cell densities (>109 cells/mL) and controlled density distributions; (3) ability to print tissues with multiscale cellular architectures and control tissues’ anisotropic properties without using scaffolds; and (4) high biocompatibility (>95% viability). With these advantages, the proposed acoustofluidic 3D bioprinting technology has the potential to significantly exceed current standards and address unmet needs in the 3D bioprinting field. We expect that our acoustofluidic 3D bioprinting technology will be of tremendous value to biomedical research communities working on fundamental in vitro and in vivo studies, cancer research, cell-cell interaction studies, tissue engineering, regenerative medicine, and drug screening.

项目概要 三维（3D）生物打印是一项快速新兴的技术，具有快速打印的潜力 近年来，在改造传统的功能性生物组织方面取得了很大进展。 然而，它们打印的组织通常缺乏分辨率和复杂性。 实现天然植物的基本功能、生理条件和各向异性特性所必需的 因此，迫切需要开发能够解决组织问题的下一代 3D 生物打印仪器。 以下三个关键技术限制：（1）无法控制打印矩阵中的内部单元位置 材料体素并实现所需的细胞间距（即细胞邻近分辨率 < 10 µm），这对于 从细胞水平确保适当的组织功能并研究 3D 中的细胞间相互作用 微环境；(2) 无法打印具有高局部细胞密度（>109 个细胞/mL）的组织，如 体内；（3）无法使用多尺度仿生细胞进行大规模组织的无支架打印 架构（例如，细胞图案和对齐），这对于实现所需的各向异性组织至关重要 在过去的十年里，我们已经掌握了依赖于多尺度细胞排列的特性和关键功能。 开发了一系列声流控（即声学与微流控的融合）技术， 特别是，我们最近开发了声流控技术。 全息术，一种基于声学、生物相容性和高分辨率的细胞操纵技术，允许 在此基础上，在聚合之前对细胞接种的基质材料进行图案化、旋转和浓缩。 技术，在这个 R01 项目中，我们建议开发和验证声流控 3D 生物打印原型 打印具有高细胞邻近分辨率（<10 µm）和复杂特征（例如仿生）的功能组织 细胞结构、受控的各向异性特性和高细胞密度）在生物相容性、快速和 我们的声流控 3D 生物打印机将通过打印血管化肿瘤球体进行验证。 基质和各向异性、神经支配、血管化的骨骼肌组织与当前的 3D 生物打印相比。 仪器，我们的声流控3D生物打印机将具有多重优势，包括：（1）能够控制内部 打印矩阵体素的细胞位置并实现高细胞邻近分辨率（< 10 µm）；（2）打印能力 具有高局部细胞密度（>109 个细胞/mL）和受控密度分布的组织；（3）打印组织的能力； 具有多尺度细胞结构，无需使用支架即可控制组织的各向异性； (4)高生物相容性(>95%的生存能力)，提出了声流控3D生物打印。 技术有潜力显着超越当前标准并解决 3D 领域未满足的需求 我们预计我们的声流控 3D 生物打印技术将为生物打印领域带来巨大价值。 致力于基础体外和体内研究、癌症研究、细胞间研究的生物医学研究团体 相互作用研究、组织工程、再生医学和药物筛选。