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

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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.

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Acknowledgements

This study was supported under the RIE2020 Industry Alignment Fund—Industry Collaboration Projects (IAF-ICP) Funding Initiative, as well as cash and in-kind contribution from the industry partner, HP Inc., through the HP-NTU Digital Manufacturing Corporate Lab. We would also like to acknowledge and thank the D300e HP team for supplying the cell-dispensing cassettes for the experiments. Wei Long Ng would like to acknowledge support from NTU Presidential Postdoctoral Fellowship.

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Authors and Affiliations

  1. HP-NTU Digital Manufacturing Corporate Lab, Nanyang Technological University (NTU), Singapore, 637460, Singapore

    Wei Long Ng, Xi Huang, Ratima Suntornnond & Wai Yee Yeong

  2. Singapore Centre for 3D Printing (SC3DP), School of Mechanical and Aerospace Engineering, Nanyang Technological University (NTU), Singapore, 639798, Singapore

    Wei Long Ng & Wai Yee Yeong

  3. HP Inc., Palo Alto, CA, 94304, USA

    Viktor Shkolnikov

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WLN: conceptualization, methodology, investigation, visualization, writing—review & editing, and supervision. XH: investigation and writing.VS: investigation and writing. RS: investigation and visualization. WYY: funding acquisition and review.

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Ng, W.L., Huang, X., Shkolnikov, V. et al. Polyvinylpyrrolidone-based bioink: influence of bioink properties on printing performance and cell proliferation during inkjet-based bioprinting. Bio-des. Manuf. 6, 676–690 (2023). https://doi.org/10.1007/s42242-023-00245-3

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