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Bio-Design and Manufacturing  2023 Vol.6 No.6 P.676-690

http://doi.org/10.1007/s42242-023-00245-3


Polyvinylpyrrolidone-based bioink: influence of bioink properties on printing performance and cell proliferation during inkjet-based bioprinting


Author(s):  Wei Long Ng, Xi Huang, Viktor Shkolnikov, Ratima Suntornnond & Wai Yee Yeong

Affiliation(s):  HP-NTU Digital Manufacturing Corporate Lab, Nanyang Technological University (NTU), Singapore 637460, Singapore; more

Corresponding email(s):   ng.wl@ntu.edu.sg, wyyeong@ntu.edu.sg

Key Words:  Biofabrication, 3D bioprinting, Drop-on-demand bioprinting, Bioink properties, Polyvinylpyrrolidone


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Wei Long Ng, Xi Huang, Viktor Shkolnikov, Ratima Suntornnond & Wai Yee Yeong. Polyvinylpyrrolidone-based bioink: influence of bioink properties on printing performance and cell proliferation during inkjet-based bioprinting[J]. Journal of Zhejiang University Science D, 2023, 6(6): 676-690.

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Abstract: 
Among the different bioprinting techniques, the drop-on-demand (DOD) jetting-based bioprinting approach facilitates contactless deposition of pico/nanoliter droplets of materials and cells for optimal cell‒matrix and cell‒cell interactions. Although bioinks play a critical role in the bioprinting process, there is a poor understanding of the influence of bioink properties on printing performance (such as filament elongation, formation of satellite droplets, and droplet splashing) and cell health (cell viability and proliferation) during the DOD jetting-based bioprinting process. An inert polyvinylpyrrolidone (PVP360, molecular weight=360 kDa) polymer was used in this study to manipulate the physical properties of the bioinks and investigate the influence of bioink properties on printing performance and cell health. Our experimental results showed that a higher bioink viscoelasticity helps to stabilize droplet filaments before rupturing from the nozzle orifice. The highly stretched droplet filament resulted in the formation of highly aligned “satellite droplets,” which minimized the displacement of the satellite droplets away from the predefined positions. Next, a significant increase in the bioink viscosity facilitated droplet deposition on the wetted substrate surface in the absence of splashing and significantly improved the accuracy of the deposited main droplet. Further analysis showed that cell-laden bioinks with higher viscosity exhibited higher measured average cell viability (%), as the presence of polymer within the printed droplets provides an additional cushioning effect (higher energy dissipation) for the encapsulated cells during droplet impact on the substrate surface, improves the measured average cell viability even at higher droplet impact velocity and retains the proliferation capability of the printed cells. Understanding the influence of bioink properties (e.g., bioink viscoelasticity and viscosity) on printing performance and cell proliferation is important for the formulation of new bioinks, and we have demonstrated precise DOD deposition of living cells and fabrication of tunable cell spheroids (nL‒µL range) using multiple types of cells in a facile manner.

南洋理工大学Wei Long Ng | 聚乙烯吡咯烷酮生物墨水:基于喷墨的生物打印过程中生物墨水特性对打印性能和细胞增殖的影响

本研究论文聚焦基于喷墨的生物打印过程中生物墨水特性对打印性能和细胞增殖的影响。在生物打印技术中,基于按需(DOD)形成液滴的生物打印方法可以实现微升/纳升的载细胞液滴的非接触式沉积,从而促进细胞-基质和细胞-细胞之间的相互作用。尽管生物墨水在打印过程中起着至关重要的作用,但研究者对于生物墨水对打印性能(液滴拉伸,卫星液滴的形成以及液滴溅射)和细胞健康(存活率和增殖)的影响仍是知之甚少的。本研究中使用惰性聚乙烯吡咯烷酮(PVP360,摩尔质量360 kDa)聚合物来调节生物墨水的物理特性,并借此研究生物墨水特性对打印质量和细胞健康的影响。实验结果表明生物墨水的粘弹性有助于稳定丝从喷嘴处的断裂过程。高度拉伸的丝导致了排列整齐的卫星液滴的形成,最大限度地减少了卫星液滴相对预定位置的偏移。其次,生物墨水的粘度在避免了液滴溅射的同时显著提高促进了液滴在湿基底的沉积精度。进一步分析表明高粘度的含细胞生物墨水由于为载细胞液滴撞击基底表面时提供额外缓冲(更高的能量消耗),而展示出较高的细胞存活率和细胞增殖能力。了解生物墨水的特性(如生物墨水的粘弹性和粘度)对打印性能和细胞增殖的影响对新型生物墨水的研发至关重要,我们已经展示了基于活细胞的精准DOD沉积,并利用简便的方式制备了含有多细胞体积可调(nL–µL)的细胞球。

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