3D bioprinted hydrogel model incorporating β-tricalcium phosphate for calcified cartilage tissue engineering
3D bioprinted hydrogel model incorporating β-tricalcium phosphate for calcified cartilage tissue engineering
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DOI：
10.1088/1758.5090/ab15cb
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发表时间：
2019-07-01
期刊：
影响因子：
9
通讯作者：
Swieszkowski, Wojciech
中科院分区：
文献类型：
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作者：
Kosik-Koziol, Alicja;Costantini, Marco;Swieszkowski, Wojciech
重建骨软骨缺损的一种有前途的策略依赖于由透明软骨、钙化软骨和软骨下骨组成的3D生物打印三区结构。到目前为止，一些研究已经在体外进行了透明软骨或骨的再生，尽管它在骨软骨组织中起着关键作用，但只有少数研究针对钙化层。在这项工作中，我们提出了一个三维仿生水凝胶支架含有β-磷酸三钙（TCP）的工程钙化软骨通过同轴针系统实施基于挤出的生物打印过程。经过彻底的生物墨水优化，我们发现0.5%w/v TCP是形成具有高形状保真度的稳定支架的最佳浓度，并具有与钙化软骨发育相关的生物学特性。特别是，我们研究了由陶瓷纳米颗粒诱导的生物打印的骨髓源性人间充质干细胞在水凝胶支架中的分化能力，所述水凝胶支架培养至21 din软骨形成培养基。为了证实所提出的方法产生钙化软骨组织的功能性体外模型的潜力，我们通过RT-qPCR和荧光免疫细胞化学定性地评估了相关软骨形成（COL 1、COL 2、COL 10A 1、ACAN）和成骨（ALPL、BGCL 2）基因标记物的定量基因表达。
One promising strategy to reconstruct osteochondral defects relies on 3D bioprinted three-zonal structures comprised of hyaline cartilage, calcified cartilage, and subchondral bone. So far, several studies have pursued the regeneration of either hyaline cartilage or bone in vitro while-despite its key role in the osteochondral region-only few of them have targeted the calcified layer. In this work, we present a 3D biomimetic hydrogel scaffold containing beta-tricalcium phosphate (TCP) for engineering calcified cartilage through a co-axial needle system implemented in extrusion-based bioprinting process. After a thorough bioink optimization, we showed that 0.5% w/v TCP is the optimal concentration forming stable scaffolds with high shape fidelity and endowed with biological properties relevant for the development of calcified cartilage. In particular, we investigate the effect induced by ceramic nano-particles over the differentiation capacity of bioprinted bone marrow-derived human mesenchymal stem cells in hydrogel scaffolds cultured up to 21 din chondrogenic media. To confirm the potential of the presented approach to generate a functional in vitro model of calcified cartilage tissue, we evaluated quantitatively gene expression of relevant chondrogenic (COL1, COL2, COL10A1, ACAN) and osteogenic (ALPL, BGLAP) gene markers by means of RT-qPCR and qualitatively by means of fluorescence immunocytochemistry.