Centre for Translational Bone, Joint and Soft Tissue Research, Faculty of Medicine, TU Dresden, 01307 Dresden, Germany.
ACS Appl Mater Interfaces. 2020 Mar 18;12(11):12557-12572. doi: 10.1021/acsami.0c00710. Epub 2020 Mar 5.
Extrusion-based bioprinting, also known as 3D bioplotting, is a powerful tool for the fabrication of tissue equivalents with spatially defined cell distribution. Even though considerable progress has been made in recent years, there is still a lack of bioinks which enable a tissue-like cell response and are plottable at the same time with good shape fidelity. Herein, we report on the development of a bioink which includes fresh frozen plasma from full human blood and thus a donor/patient-specific protein mixture. By blending of the plasma with 3 w/v% alginate and 9 w/v% methylcellulose, a pasty bioink (plasma-alg-mc) was achieved, which could be plotted with high accuracy and furthermore allowed bioplotted mesenchymal stromal cells (MSC) and primary osteoprogenitor cells to spread within the bioink. In a second step, the novel plasma-based bioink was combined with a plottable self-setting calcium phosphate cement (CPC) to fabricate bone-like tissue constructs. The CPC/plasma-alg-mc biphasic constructs revealed open porosity over the entire time of cell culture (35 d), which is crucial for bone tissue engineered grafts. The biphasic structures could be plotted in volumetric and clinically relevant dimensions and complex shapes could be also generated, as demonstrated for a scaphoid bone model. The plasma bioink potentiated that bioplotted MSC were not harmed by the setting process of the CPC. Latest after 7 days, MSC migrated from the hydrogel to the CPC surface, where they proliferated to 20-fold of the initial cell number covering the entire plotted constructs with a dense cell layer. For bioplotted and osteogenically stimulated osteoprogenitor cells, a significantly increased alkaline phosphatase activity was observed in CPC/plasma-alg-mc constructs in comparison to plasma-free controls. In conclusion, the novel plasma-alg-mc bioink is a promising new ink for several forms of bioprinted tissue equivalents and especially gainful for the combination with CPC for enhanced, biofabricated bone-like constructs.
基于挤出的生物打印，也称为 3D 生物绘图，是制造具有空间定义细胞分布的组织等效物的有力工具。尽管近年来取得了相当大的进展，但仍然缺乏能够实现类似组织的细胞反应并且能够以良好的形状保真度同时进行绘图的生物墨水。在此，我们报告了一种生物墨水的开发，该生物墨水包括来自全人血的新鲜冷冻血浆，从而形成了供体/患者特异性蛋白质混合物。通过将血浆与 3 w/v%藻酸盐和 9 w/v%甲基纤维素混合，获得了一种糊状生物墨水（血浆-藻酸盐-mc），可以高精度绘制，并且还允许生物绘制的间充质基质细胞（MSC）和原代成骨前体细胞在生物墨水中扩散。在第二步中，将新型基于血浆的生物墨水与可绘制的自凝固磷酸钙水泥（CPC）结合，以制造类骨组织构建体。CPC/血浆-藻酸盐-mc 双相构建体在整个细胞培养过程中（35 天）均显示出开放孔隙率，这对于骨组织工程移植物至关重要。双相结构可以以体积和临床相关的尺寸进行绘制，并且可以生成复杂的形状，如手舟骨模型所示。该血浆生物墨水增强了生物绘制的 MSC 不会被 CPC 的凝固过程所伤害。在第 7 天之后，MSC 从水凝胶迁移到 CPC 表面，在那里它们增殖到初始细胞数的 20 倍，覆盖整个绘制的构建体并形成致密的细胞层。对于生物绘制和成骨刺激的成骨前体细胞，与无血浆对照相比，在 CPC/血浆-藻酸盐-mc 构建体中观察到碱性磷酸酶活性显著增加。总之，新型血浆-藻酸盐-mc 生物墨水是几种形式的生物打印组织等效物的有前途的新型墨水，特别是对于与 CPC 结合以增强生物制造的类骨构建体特别有益。
