Nishiguchi Akihiro, Kapiti Gent, Höhner J Robin, Singh Smriti, Moeller Martin
DWI-Leibniz-Institute for Interactive Materials, Forckenbeckstrasse 50, D-52056 Aachen Germany.
ACS Appl Bio Mater. 2020 Apr 20;3(4):2378-2384. doi: 10.1021/acsabm.0c00116. Epub 2020 Mar 31.
Living tissues dynamically modulate their structure and functions through physical and biochemical interactions in the three-dimensional (3D)-microenvironment for their homeostasis or the developmental process of an embryo. However, the manipulation of cellular functions in vitro is still challenging due to the lack of a dynamic material system that can vary the 3D-cellular microenvironment in time and space. Here, we show an in situ 3D-printing technique based on multiphoton lithography using a biocompatible photoresist, bio-ink. The bio-ink composed of protein-photosensitizer conjugates has the ability to cause singlet oxygen and cross-linking reaction to fabricate protein gels with submicrometer-scale precision. Remarkably, the conjugates substantially improve the cytocompatibility and the efficiency of gelation due to the stealth effect of rose bengal (RB) and efficient transfer of singlet oxygen to bovine serum albumin (BSA). 3D-printing in the presence of cells allows for the microfabrication of a protein scaffold and controlled single-cell behavior. This dynamic material system to direct cell fate may offer emerging applications for drug discovery and regenerative medicine.
活组织通过三维（3D）微环境中的物理和生化相互作用动态调节其结构和功能，以维持自身稳态或胚胎的发育过程。然而，由于缺乏能够在时间和空间上改变3D细胞微环境的动态材料系统，体外细胞功能的操控仍然具有挑战性。在此，我们展示了一种基于多光子光刻的原位3D打印技术，该技术使用生物相容性光致抗蚀剂——生物墨水。由蛋白质 - 光敏剂共轭物组成的生物墨水能够引发单线态氧和交联反应，以亚微米级精度制造蛋白质凝胶。值得注意的是，由于孟加拉玫瑰红（RB）的隐身效应以及单线态氧向牛血清白蛋白（BSA）的有效转移，共轭物显著提高了细胞相容性和凝胶化效率。在细胞存在的情况下进行3D打印，可以实现蛋白质支架的微制造和对单细胞行为的控制。这种用于指导细胞命运的动态材料系统可能为药物发现和再生医学提供新的应用。
