Trinity Centre for Bioengineering, Trinity Biomedical Sciences Institute, Trinity College Dublin, Ireland; Department of Mechanical and Manufacturing Engineering, School of Engineering, Trinity College Dublin, Ireland; Advanced Materials and Bioengineering Research Centre (AMBER), Trinity College Dublin and Royal College of Surgeons, Ireland.
Trinity Centre for Bioengineering, Trinity Biomedical Sciences Institute, Trinity College Dublin, Ireland; Department of Mechanical and Manufacturing Engineering, School of Engineering, Trinity College Dublin, Ireland.
J Control Release. 2019 May 10;301:13-27. doi: 10.1016/j.jconrel.2019.03.006. Epub 2019 Mar 8.
The regeneration of complex tissues and organs remains a major clinical challenge. With a view towards bioprinting such tissues, we developed a new class of pore-forming bioink to spatially and temporally control the presentation of therapeutic genes within bioprinted tissues. By blending sacrificial and stable hydrogels, we were able to produce bioinks whose porosity increased with time following printing. When combined with amphipathic peptide-based plasmid DNA delivery, these bioinks supported enhanced non-viral gene transfer to stem cells in vitro. By modulating the porosity of these bioinks, it was possible to direct either rapid and transient (pore-forming bioinks), or slower and more sustained (solid bioinks) transfection of host or transplanted cells in vivo. To demonstrate the utility of these bioinks for the bioprinting of spatially complex tissues, they were next used to zonally position stem cells and plasmids encoding for either osteogenic (BMP2) or chondrogenic (combination of TGF-β3, BMP2 and SOX9) genes within networks of 3D printed thermoplastic fibers to produce mechanically reinforced, gene activated constructs. In vivo, these bioprinted tissues supported the development of a vascularised, bony tissue overlaid by a layer of stable cartilage. When combined with multiple-tool biofabrication strategies, these gene activated bioinks can enable the bioprinting of a wide range of spatially complex tissues.
复杂组织和器官的再生仍然是一个主要的临床挑战。为了实现这些组织的生物打印，我们开发了一类新的孔形成生物墨水，以在生物打印的组织内空间和时间上控制治疗基因的呈现。通过混合牺牲性和稳定的水凝胶，我们能够生产出在打印后随时间增加其孔隙率的生物墨水。当与基于两亲肽的质粒 DNA 递药结合使用时，这些生物墨水支持体外干细胞的非病毒基因转染增强。通过调节这些生物墨水的孔隙率，可以在体内定向快速且短暂（孔形成生物墨水）或较慢且更持续（固体生物墨水）转染宿主或移植细胞。为了证明这些生物墨水在空间复杂组织的生物打印中的实用性，接下来将它们用于在 3D 打印热塑性纤维网络内对干细胞和编码成骨（BMP2）或软骨形成（TGF-β3、BMP2 和 SOX9 的组合）基因的质粒进行分区定位，以产生具有机械增强功能和基因激活的构建体。在体内，这些生物打印的组织支持血管化骨组织的形成，其上覆盖着一层稳定的软骨。当与多种工具生物制造策略结合使用时，这些基因激活的生物墨水可以实现广泛的空间复杂组织的生物打印。
