Electron Spin Resonance Probe Incorporation into Bioinks Permits Longitudinal Oxygen Imaging of Bioprinted Constructs.

IF 3 4区医学 Q2 RADIOLOGY, NUCLEAR MEDICINE & MEDICAL IMAGING

Molecular Imaging and Biology Pub Date : 2024-06-01 Epub Date: 2023-12-01 DOI:10.1007/s11307-023-01871-0

Sajad Sarvari, Duncan McGee, Ryan O'Connell, Oxana Tseytlin, Andrey A Bobko, Mark Tseytlin

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引用次数: 0

Abstract

Purpose: Bioprinting is an additive manufacturing technology analogous to 3D printing. Instead of plastic or resin, cell-laden hydrogels are used to produce a construct of the intended biological structure. Over time, cells transform this construct into a functioning tissue or organ. The process of printing followed by tissue maturation is referred to as 4D bioprinting. The fourth dimension is temporal. Failure to provide living cells with sufficient amounts of oxygen at any point along the developmental timeline may jeopardize the bioprinting goals. Even transient hypoxia may alter cells' differentiation and proliferation or trigger apoptosis. Electron paramagnetic resonance (EPR) imaging modality is proposed to permit 4D monitoring of oxygen within bioprinted structures.

Procedures: Lithium octa-n-butoxy-phthalocyanine (LiNc-BuO) probes have been introduced into gelatin methacrylate (GelMA) bioink. GelMA is a cross-linkable hydrogel, and LiNc-BuO is an oxygen-sensitive compound that permits longitudinal oximetric measurements. The effects of the oxygen probe on printability have been evaluated. A digital light processing (DLP) bioprinter was built in the laboratory. Bioprinting protocols have been developed that consider the optical properties of the GelMA/LiNc-BuO composites. Acellular and cell-laden constructs have been printed and imaged. The post-printing effect of residual photoinitiator on oxygen depletion has been investigated.

Results: Models have been successfully printed using a lab-built bioprinter. Rapid scan EPR images reflective of the expected oxygen concentration levels have been acquired. An unreported problem of oxygen depletion in bioprinted constructs by the residual photoinitiator has been documented. EPR imaging is proposed as a control method for its removal. The oxygen consumption rates by HEK293T cells within a bioprinted cylinder have been imaged and quantified.

Conclusions: The feasibility of the cointegration of 4D EPR imaging and 4D bioprinting has been demonstrated. The proof-of-concept experiments, which were conducted using oxygen probes loaded into GelMA, lay the foundation for a broad range of applications, such as bioprinting with many types of bioinks loaded with diverse varieties of molecular spin probes.

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电子自旋共振探针纳入生物墨水允许纵向氧成像的生物打印结构。

目的:生物打印是一种类似于3D打印的增材制造技术。而不是塑料或树脂，细胞负载的水凝胶被用来产生预期的生物结构的结构。随着时间的推移，细胞将这种结构转化为一个功能性的组织或器官。组织成熟后的打印过程被称为4D生物打印。第四个维度是暂时的。如果不能在发育过程中的任何时刻为活细胞提供足够的氧气，可能会危及生物打印的目标。即使是短暂的缺氧也可能改变细胞的分化和增殖或引发细胞凋亡。电子顺磁共振(EPR)成像模式提出，允许4D监测氧气在生物打印结构。程序:锂八正丁氧基酞菁(LiNc-BuO)探针已引入明胶甲基丙烯酸酯(GelMA)生物墨水。GelMA是一种可交联的水凝胶，而LiNc-BuO是一种氧敏感化合物，允许纵向氧饱和度测量。评价了氧探针对印刷适性的影响。在实验室中建立了数字光处理(DLP)生物打印机。考虑到GelMA/LiNc-BuO复合材料的光学特性，生物打印方案已经开发出来。无细胞和载细胞结构已被打印和成像。研究了残留光引发剂对印后耗氧的影响。结果:使用实验室建造的生物打印机成功打印了模型。快速扫描EPR图像反映预期的氧浓度水平已经获得。一个未报道的问题，氧气消耗在生物打印结构的残留光引发剂已被记录。提出了EPR成像作为一种控制方法，以消除其。HEK293T细胞在生物打印圆柱体内的耗氧率已被成像和量化。结论:4D EPR成像与4D生物打印协整的可行性已得到证实。概念验证实验使用装载在GelMA中的氧探针进行，为广泛的应用奠定了基础，例如使用装载不同种类分子自旋探针的多种生物墨水进行生物打印。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Molecular Imaging and Biology 医学-核医学

CiteScore

6.90

自引率

3.20%

发文量

审稿时长

3 months

期刊介绍： Molecular Imaging and Biology (MIB) invites original contributions (research articles, review articles, commentaries, etc.) on the utilization of molecular imaging (i.e., nuclear imaging, optical imaging, autoradiography and pathology, MRI, MPI, ultrasound imaging, radiomics/genomics etc.) to investigate questions related to biology and health. The objective of MIB is to provide a forum to the discovery of molecular mechanisms of disease through the use of imaging techniques. We aim to investigate the biological nature of disease in patients and establish new molecular imaging diagnostic and therapy procedures. Some areas that are covered are: Preclinical and clinical imaging of macromolecular targets (e.g., genes, receptors, enzymes) involved in significant biological processes. The design, characterization, and study of new molecular imaging probes and contrast agents for the functional interrogation of macromolecular targets. Development and evaluation of imaging systems including instrumentation, image reconstruction algorithms, image analysis, and display. Development of molecular assay approaches leading to quantification of the biological information obtained in molecular imaging. Study of in vivo animal models of disease for the development of new molecular diagnostics and therapeutics. Extension of in vitro and in vivo discoveries using disease models, into well designed clinical research investigations. Clinical molecular imaging involving clinical investigations, clinical trials and medical management or cost-effectiveness studies.