3D-Bioprinting of Polylactic Acid (PLA) Nanofiber-Alginate Hydrogel Bioink Containing Human Adipose-Derived Stem Cells
3D-Bioprinting of Polylactic Acid (PLA) Nanofiber-Alginate Hydrogel Bioink Containing Human Adipose-Derived Stem Cells
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DOI：
10.1021/acsbiomaterials.6b00196
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发表时间：
2016-10-01
影响因子：
5.8
通讯作者：
Shirwaiker, Rohan A.
中科院分区：
文献类型：
--
作者：
Narayanan, Lokesh Karthik;Huebner, Pedro;Shirwaiker, Rohan A.
生物墨水通过提供支持环境在3D生物打印中发挥核心作用，在该支持环境中，封装的细胞可以承受数字驱动制造过程中遇到的应力，并继续成熟，增殖并最终形成细胞外基质（ECM）。为了最有效，重要的是生物打印的构建体尽可能接近地重现天然组织环境。因此，肌肉骨骼软组织构建体可以受益于模拟其纳米纤维基质构成的生物墨水，这对其功能也至关重要。这项研究的重点是开发和概念验证评估的纤维生物墨水组成的藻酸盐水凝胶，聚乳酸纳米纤维，和人类脂肪来源的干细胞（hASC）的生物打印这样的组织结构。首先，在体外16天内在3D生物作图链中评估hASC增殖和活力。然后，使用磁共振图像数字建模的人类内侧膝关节半月板被生物打印并在体外评估8周。结果表明，纳米纤维增强的生物墨水允许生物打印链内更高水平的细胞增殖，在第7天达到峰值，同时在第16天仍保持绝大多数活细胞。与没有纳米纤维的生物墨水相比，该生物墨水中第7天的细胞代谢活性高28.5%。生物打印半月板在第4周和第8周的组织学显示，在结构的外部区域与内部区域中，细胞密度分别高出54%和147%。在hASC周围的区域中也注意到胶原蛋白和蛋白聚糖的存在，表明ECM分泌和软骨形成分化。
Bioinks play a central role in 3D-bioprinting by providing the supporting environment within which encapsulated cells can endure the stresses encountered during the digitally driven fabrication process and continue to mature, proliferate, and eventually form extracellular matrix (ECM). In order to be most effective, it is important that bioprinted constructs recapitulate the native tissue milieu as closely as possible. As such, musculoskeletal soft tissue constructs can benefit from bioinks that mimic their nanofibrous matrix constitution, which is also critical to their function. This study focuses on the development and proof-of-concept assessment of a fibrous bioink composed of alginate hydrogel, polylactic acid nanofibers, and human adipose-derived stem cells (hASC) for bioprinting such tissue constructs. First, hASC proliferation and viability were assessed in 3D-bioplotted strands over 16 days in vitro. Then, a human medial knee meniscus digitally modeled using magnetic resonance images was bioprinted and evaluated over 8 weeks in vitro. Results show that the nanofiber-reinforced bioink allowed higher levels of cell proliferation within bioprinted strands, with a peak at day 7, while still maintaining a vast majority of viable cells at day 16. The cell metabolic activity on day 7 was 28.5% higher in this bioink compared to the bioink without nanofibers. Histology of the bioprinted meniscus at both 4 and 8 weeks showed 54% and 147% higher cell density, respectively, in external versus internal regions of the construct. The presence of collagen and proteoglycans was also noted in areas surrounding the hASC, indicating ECM secretion and chondrogenic differentiation.