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Bioink formulations to ameliorate bioprinting-induced loss of cellular viability.
Biointerphases ( IF 2.1 ) Pub Date : 2019-10-16 , DOI: 10.1116/1.5111392
Sudipto Datta 1 , Ankita Das 1 , Amit Roy Chowdhury 1 , Pallab Datta 1
Affiliation  

Extrusion bioprinting, the most affordable and convenient bioprinting modality, is also associated with high process-induced cell deaths. Mechanical stresses on the cells during pneumatic or piston extrusion generate excessive reactive oxygen species and activate apoptosis, inflammatory pathways in the cells. In this study, a bioink formulation is augmented with an antioxidant, N-acetyl cysteine (NAC) as a possible solution to abrogate the effect of bioprinting-associated cell survival losses. The NAC addition to bioinks did not affect the bioprinting process, shape fidelity, or the mechanical properties of the constructs to any large extent. However, the bioprinting process conducted at 0.30 MPa pressure and 410 μm nozzle inner diameter with bioinks of 3% w/v alginate, 105 cells/ml resulted in survival losses of up to 25% for MC3T3 cells. In contrast, NAC bioinks showed a significant (p < 0.01) improvement in day 1 cell survival (91%), while the enhancement in day 3 cell viability was still greater. It was further observed that the reactive oxygen species (ROS) load of bioprinted constructs was approximately 1.4 times higher compared to control, whereas NAC containing constructs reduced the ROS load at levels comparable to control samples. The effect on apoptosis and inflammation markers showed that NAC had a greater role in modulating apoptosis. It is concluded that the presented approach to preserve cell viability and functionality would be advantageous over other contemporary methods (like alterations in extrusion pressure, nozzle diameter, polymer concentration, etc.) as viability can be preserved without compromising the fabrication time or the resolution/mechanical properties of the constructs with this bioink formulation approach.

中文翻译:

Bioink制剂可改善生物印刷引起的细胞活力丧失。

挤出生物打印是最实惠,最方便的生物打印方式,也与高过程诱导的细胞死亡有关。在气动或活塞挤压过程中,细胞上的机械应力会产生过多的活性氧,并激活细胞中的凋亡,炎症途径。在这项研究中,生物墨水配方中添加了抗氧化剂N-乙酰半胱氨酸(NAC)作为消除与生物印刷相关的细胞存活损失的影响的可能解决方案。NAC添加到生物墨水中不会在很大程度上影响生物印刷过程,形状保真度或构建体的机械性能。但是,生物打印过程是在0.30 MPa压力和410μm喷嘴内径下进行的,其中生物墨水为3%w / v藻酸盐,105个细胞/ ml,导致MC3T3细胞的生存损失高达25%。相比之下,NAC生物墨水在第1天的细胞存活率中显示了显着的改善(p <0.01)(91%),而在第3天的细胞存活率中的增强作用更大。进一步观察到,与对照相比,生物打印构建物的活性氧(ROS)负载约高1.4倍,而含NAC的构建物降低了ROS负载,其水平与对照样品相当。对细胞凋亡和炎症标志物的影响表明,NAC在调节细胞凋亡中具有更大的作用。结论是,提出的保持细胞活力和功能性的方法将优于其他当代方法(例如改变挤出压力,喷嘴直径,聚合物浓度等)。
更新日期:2019-11-01
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