3D bioprinting of anisotropic engineered tissue constructs with ultrasonically induced cell patterning
3D bioprinting of anisotropic engineered tissue constructs with ultrasonically induced cell patterning
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DOI：
10.1016/j.addma.2020.101042
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发表时间：
2020-03-01
影响因子：
11
通讯作者：
Shirwaiker, Rohan
中科院分区：
文献类型：
--
作者：
Chansoria, Parth;Shirwaiker, Rohan
随着3D生物打印继续发展成为体外工程复杂人体组织的一种有前途的替代方案，需要增强生物打印过程以实现原始组织中发现的必要的细胞和细胞外组织特征。虽然生物墨水内的细胞分布通常是均匀的，但在生物打印的构建体内掺入适当的细胞图案化是最终形成对其生物力学形式和功能至关重要的各向异性组织的组织基质的必要的第一步。这项研究描述了一种新的生物打印技术，该技术使用超声驻波体声波（SBAW）将细胞组织成粘性生物墨水内的可控各向异性图案，同时保持高细胞活力。首先，我们开发了一个三维计算模型来识别SBAW压力模式响应于多个超声波频率（0.71-2 MHz）。然后，我们实验分析了人脂肪源性干细胞（hASC）和人骨肉瘤细胞（MG 63）在藻酸盐中的模式和活力作为SBAW频率的函数。计算结果表明，形成平行的压力股附近的沉积层的底部，这是证实了细胞图案的实验图像具有较高的压力振幅。发现链间间距受到频率的影响（p < 0.0001），而细胞类型和频率之间的相互作用效应决定了链的宽度（p = 0.02）。此外，我们证明了协同生物打印和SBAW诱导的图案化的hASC内藻酸盐和明胶甲基丙烯酸酯（GelMA）的建设串联化学和光交联，分别。在整个实验设计空间中，在藻酸盐和GelMA构建体中注意到至少80%的相关细胞图案化和活力。最后，我们展示了3层GelMA构建体的基于瓮光聚合的生物打印，其中hASC链铺设模式在层间为0-45-90度。这项工作代表了推进生物打印能力以实现仿生组织构建的一步。
As 3D bioprinting continues to evolve as a promising alternative to engineer complex human tissues in-vitro, there is a need to augment bioprinting processes to achieve the requisite cellular and extracellular organizational characteristics found in the original tissues. While the cell distribution within bioinks is typically homogeneous, incorporating appropriate cellular patterning within the bioprinted constructs is an essential first step towards the eventual formation of anisotropically organized tissue matrix essential to its biomechanical form and function. This study describes a new bioprinting technique that uses ultrasonic standing bulk acoustic waves (SBAW) to organize cells into controllable anisotropic patterns within viscous bioinks while maintaining high cell viability. First, we develop a 3D computational model to discern the SBAW pressure pattern in response to multiple ultrasonic frequencies (0.71-2 MHz). We then experimentally analyze the patterns and viabilities of human adipose-derived stem cells (hASC) and human osteosarcoma cells (MG63) in alginate as a function of the SBAW frequency. Computational results indicate the formation of parallel pressure strands with higher pressure amplitudes near the bottom of the deposited layer, which is corroborated by experimental images of cell patterning. The inter-strand spacing is found to be affected by the frequency (p < 0.0001), while an interaction effect between the cell type and frequency governs the width of the strands (p = 0.02). Further, we demonstrate the synergistic bioprinting and SBAW-induced patterning of hASC within alginate and gelatin methacrylate (GelMA) constructs in tandem with chemical and photo-crosslinking, respectively. Pertinent cellular patterning and viability of at least 80 % were noted in the alginate and GelMA constructs across the experimental design space. Finally, we demonstrate the vat photo-polymerization-based bioprinting of a 3-layered GelMA construct with hASC strand lay pattern of 0-45-90 degrees across the layers. This work represents a step forward in advancing bioprinting capabilities to achieve biomimetic tissue constructs.