Collaborative Research: 4D Bioprinting of Near-infrared Light Responsive Smart Constructs for Pluripotent Stem Cell Derived Cardiomyocyte Engineering

合作研究：用于多能干细胞衍生心肌细胞工程的近红外光响应智能结构的 4D 生物打印

• 批准号: 1856321
• 负责人: Lijie Grace Zhang
• 金额: $ 27.5万
• 依托单位: George Washington University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1856321&HistoricalAwards=false
• 关键词: Collaborative; Research; 4D; Bioprinting; Near

4D bioprinting is an emerging manufacturing process to create smart tissue constructs where cellular behaviors can be regulated in both space and time. The objective of this study is to 4D bioprint dynamic light-responsive smart constructs to control the functions of cardiomyocytes (heart muscle cells) derived from human induced pluripotent stem cells. For this purpose, novel light-sensitive ink materials will be synthesized and characterized, and a series of smart structures will be printed to study the effect of the bioprinting process on the structure's dynamic shape changes. Finally, the effects of light-triggered 4D structures on regulating cardiomyocyte growth, differentiation, and beating will be explored. The manipulation of cardiac cell behaviors through 4D bioprinting processes will expand our understanding of cardiac cell function for potential cardiac engineering applications. In addition, this collaborative research will lay the foundation for next-generational 4D bioprinting platforms. Educational and outreach activities will involve a collaboration between the George Washington University and University of Maryland, College Park for sharing research experiences, improvement of existing courses, and inclusion of undergraduate and K-12 students, with broad representations of underrepresented minorities in research, to help educate the future bioengineering workforce.The objective of this study is to 4D bioprint reprogrammable near-infrared light (NIR) responsive smart constructs and to discover 4D dynamic effects on controlling human induced pluripotent stem cell derived cardiomyocyte (iPSC-CM) function and beating behaviors, hypothesizing that these structures will be successfully created and the 4D dynamic effect will greatly improve iPSC-CM functionality and beating behaviors. The first key innovation focuses on creating a new generation of light-sensitive smart inks with precisely controlled multi-responsive 4D effects and bio-functionality. A human benign NIR sensitive moiety will be used in the synthesized 4D ink as a model smart switch. The long-wavelength NIR can efficiently penetrate through the printed biomaterials compared to ultraviolet/visible light and will not harm the surrounding cells. The iPSC-CM is selected because it is a mechanically responsive cell line that is perfect for studying 4D dynamic effects for basic and translational cardiovascular research. This cell line also offers the key advantages of being in virtually unlimited cardiomyocyte supply, as well as having a high regenerative capacity. The project's objectives will be accomplished under three aims. The FIRST Aim is to formulate and characterize a novel smart ink with three key components: a natural triglyceride-based monomer that will serve as the printable matrix of the ink, a liquid crystal polymer that is a critical functional component to exert the reprogrammable property and a NIR moiety with a light polymerizable double bond group. The reaction and the molecular structures will be characterized by Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR) and differential scanning calorimetry (DSC). The 4D inks will be formulated by varying the ratios of the above components in order to achieve a desired, printable rheological property. The SECOND Aim is to bioprint smart structures and explore the effect of the bioprinting process on 4D dynamic shape changes. A custom designed stereolithography (SL) bioprinter, which is capable of controlling key bioprinting parameters (printing speed, printing layer height and laser intensity), will be used to print the synthesized ink materials. The relationship between the printing parameters and the 4D shape change of the light-sensitive smart structure will be established. The THIRD aim is to investigate the dynamic shape change of NIR responsive structures on regulating iPSC-CM functions. The interaction of iPSC-CMs with the NIR responsive structures will be thoroughly studied; the growth of iPSC-CMs on the bioprinted constructs will be determined; the effect of the 4D variations of the bioprinted constructs on calcium transience, myogenesis, and gene expression of iPSC-CMs will be qualitatively and quantitatively examined by Fluo-4 AM, immunocytochemistry, and gene analysis. This study will for the first time explore the fundamental interactions between NIR regulated 4D structures and cardiomyocyte behaviors.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

4D Bioprinting是一个新兴的制造过程，可以创建智能组织构建体，其中可以在时空和时间上调节细胞行为。这项研究的目的是针对4D Bioprint动态光响应式智能构建体，以控制源自人类诱导的多能干细胞的心肌细胞（心肌细胞）的功能。 为此，将合成和表征新颖的光敏墨水材料，并将打印一系列智能结构，以研究生物印刷过程对结构动态形状变化的影响。最后，将探索光触发的4D结构对调节心肌细胞生长，分化和跳动的影响。通过4D生物打印过程对心脏细胞行为的操纵将扩展我们对潜在心脏工程应用的心脏细胞功能的理解。此外，这项合作研究将为下一代4D生物打印平台奠定基础。教育和宣传活动将涉及乔治华盛顿大学与马里兰州大学公园，分享研究经验，改善现有课程，包括本科生和K-12学生的成员，并在研究领域内部不足的少数群体代表不足的少数群体，以帮助未来的生物工程构造，以帮助未来的生物工程。为了发现4D动态影响对控制人类诱导的多能干细胞衍生的心肌细胞（IPSC-CM）功能和跳动行为，假设这些结构将成功创建，而4D动态效应将大大改善IPSC-CM功能和殴打行为。第一个关键创新的重点是创建新一代的光敏智能油墨，具有精确控制的多响应4D效果和生物功能。 人类良性NIR敏感部分将用于合成的4D墨水作为型号智能开关。与紫外线/可见光相比，长波长NIR可以有效地穿过印刷的生物材料，不会损害周围细胞。 选择IPSC-CM是因为它是一种机械响应的细胞系，非常适合研究基本和转化心血管研究的4D动态效应。 该细胞系还提供了几乎无限的心肌细胞供应以及具有较高再生能力的关键优势。该项目的目标将以三个目标来实现。 第一个目的是用三个关键组件制定和表征新型的智能墨水：一种基于天然甘油三酸酯的单体，它将用作墨水的可打印矩阵，墨水是一种液晶聚合物，是一种至关重要的功能组件，可启动可重编程的特性和具有轻度聚合聚合物的双键双键组的NIR部分。反应和分子结构将以傅立叶变换红外光谱法（FTIR），核磁共振（NMR）和差异扫描量热法（DSC）为特征。 4d墨水将通过改变上述组件的比率来制定4D墨水，以实现所需的可打印的流变特性。 第二个目的是生物生物智能结构并探索生物打印过程对4D动态形状变化的影响。定制设计的立体光刻（SL）Bioprointer，能够控制关键的生物打印参数（打印速度，打印层高度和激光强度），用于打印合成的墨水材料。将建立印刷参数与光敏智能结构的4D形状变化之间的关系。第三个目的是研究在调节IPSC-CM函数的NIR响应结构的动态形状变化。 IPSC-CM与NIR响应性结构的相互作用将得到彻底研究； IPSC-CM在生物打印构建体上的生长将确定；生物打印构建体的4D变化对IPSC-CMS钙瞬时，肌发生和基因表达的影响将通过Fluo-4 AM，免疫细胞化学和基因分析进行定性和定量检查。这项研究将首次探讨NIR管制的4D结构与心肌细胞行为之间的基本相互作用。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子和更广泛影响的评估来评估的支持标准。

项目成果

4D printing soft robotics for biomedical applications

• DOI: 10.1016/j.addma.2020.101567
• 发表时间: 2020-12-01
• 期刊: ADDITIVE MANUFACTURING
• 影响因子: 11
• 作者: Hann, Sung Yun;Cui, Haitao;Zhang, Lijie Grace
• 通讯作者: Zhang, Lijie Grace

4D printing in biomedical applications: emerging trends and technologies

• DOI: 10.1039/d1tb01335a
• 发表时间: 2021-08-09
• 期刊: JOURNAL OF MATERIALS CHEMISTRY B
• 影响因子: 7
• 作者: Agarwal, Tarun;Hann, Sung Yun;Maiti, Tapas Kumar
• 通讯作者: Maiti, Tapas Kumar

Recent advances in bioprinting technologies for engineering cardiac tissue.

用于工程心脏组织的生物打印技术的最新进展。

• DOI: 10.1016/j.msec.2021.112057
• 发表时间: 2021-05
• 期刊: MATERIALS SCIENCE & ENGINEERING C-MATERIALS FOR BIOLOGICAL APPLICATIONS
• 影响因子: 7.9
• 作者: Agarwal, Tarun;Fortunato, Gabriele Maria;Hann, Sung Yun;Ayan, Bugra;Vajanthri, Kiran Yellappa;Presutti, Dario;Cui, Haitao;Chan, Alex H. P.;Costantini, Marco;Onesto, Valentina;Di Natale, Concetta;Huang, Ngan F.;Makvandi, Pooyan;Shabani, Majid;Maiti, Tapas Kumar;Zhang, Lijie Grace;De Maria, Carmelo
• 通讯作者: De Maria, Carmelo

4D physiologically adaptable cardiac patch: A 4-month in vivo study for the treatment of myocardial infarction

• DOI: 10.1126/sciadv.abb5067
• 发表时间: 2020-06-01
• 期刊: SCIENCE ADVANCES
• 影响因子: 13.6
• 作者: Cui, Haitao;Liu, Chengyu;Zhang, Lijie Grace
• 通讯作者: Zhang, Lijie Grace