AccScience Publishing / IJB / Volume 8 / Issue 1 / DOI: 10.18063/ijb.v8i1.424
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RESEARCH ARTICLE

Controlling Droplet Impact Velocity and Droplet Volume: Key Factors to Achieving High Cell Viability in Sub-Nanoliter Droplet-based Bioprinting

Wei Long Ng1* Huang Xi1 Viktor Shkolnikov2 Guo Liang Goh3 Ratima Suntornnond1 Wai Yee Yeong1,3*
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1 HP-NTU Digital Manufacturing Corporate Lab, 65 Nanyang Avenue 637460, Singapore
2 HP Inc., 1501 Page Mill Road, Palo Alto, CA 94304, United States
3 Singapore Centre for 3D Printing (SC3DP), School of Mechanical and Aerospace Engineering, Nanyang Technological University (NTU), 50 Nanyang Avenue 639798, Singapore
Submitted: 28 July 2021 | Accepted: 16 September 2021 | Published: 28 October 2021
© 2021 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
Abstract

Three-dimensional (3D) bioprinting systems serve as advanced manufacturing platform for the precise deposition of cells and biomaterials at pre-defined positions. Among the various bioprinting techniques, the drop-on-demand jetting approach facilitates deposition of pico/nanoliter droplets of cells and materials for study of cell-cell and cell-matrix interactions. Despite advances in the bioprinting systems, there is a poor understanding of how the viability of primary human cells within sub-nanoliter droplets is affected during the printing process. In this work, a thermal inkjet system is utilized to dispense sub-nanoliter cell-laden droplets, and two key factors – droplet impact velocity and droplet volume – are identified to have significant effect on the viability and proliferation of printed cells. An increase in the cell concentration results in slower impact velocity, which leads to higher viability of the printed cells and improves the printing outcome by mitigating droplet splashing. Furthermore, a minimum droplet volume of 20 nL per spot helps to mitigate evaporation-induced cell damage and maintain high viability of the printed cells within a printing duration of 2 min. Hence, controlling the droplet impact velocity and droplet volume in sub-nanoliter bioprinting is critical for viability and proliferation of printed human primary cells.

Keywords
3D Bioprinting
3D Printing
Biofabrication
Drop-on-demand printing
Sub-nanoliter cell printing
References

1. Ng WL, Chua CK, Shen YF, 2019, Print Me An Organ! Why We Are Not There Yet. Prog Polym Sci, 97:101145. https://doi.org/10.1016/j.progpolymsci.2019.101145

2. Murphy SV, Atala A, 2014, 3D Bioprinting of Tissues and Organs. Nat Biotechnol, 32:773–85. https://doi.org/10.1038/nbt.2958

3. Lee JM, Ng WL, Yeong WY, 2019, Resolution and Shape in Bioprinting: Strategizing Towards Complex Tissue and Organ Printing. Appl Phys Rev, 6:011307. https://doi.org/10.1063/1.5053909

4. Ng WL, Chan A, Ong YS, et al., 2020, Deep Learning for Fabrication and Maturation of 3D Bioprinted Tissues and Organs. Virtual Phys Prototyp, 15:340–58. https://doi.org/10.1080/17452759.2020.1771741

5. Wan AC, 2016, Recapitulating Cell-Cell Interactions for Organoid Construction-are Biomaterials Dispensable? Trends Biotechnol, 34:711–21. https://doi.org/10.1016/j.tibtech.2016.02.015

6. Gruene M, Pflaum M, Hess C, et al., 2011, Laser Printing of Three-dimensional Multicellular Arrays for Studies of Cell-Cell and Cell-Environment Interactions. Tissue Eng Part C Methods, 17:973–82. https://doi.org/10.1089/ten.tec.2011.0185

7. Ng WL, Lee JM, Zhou M, et al., 2020, Hydrogels for 3-D Bioprinting-based Tissue Engineering. In: Narayan R, editor. Rapid Prototyping of Biomaterials. Chapel Hill, NC: Elsevier, p183-204. https://doi.org/10.1016/b978-0-08-102663-2.00008-3

8. Kathawala MH, Ng WL, Liu D, et al., 2019, Healing of Chronic Wounds an Update of Recent Developments and Future Possibilities. Tissue Eng Part B Rev, 25:429–44. https://doi.org/10.1089/ten.teb.2019.0019

9. Rose JC, De Laporte L, 2018, Hierarchical Design of Tissue Regenerative Constructs. Adv Healthc Mater, 7:1701067. https://doi.org/10.1002/adhm.201701067

10. Ng WL, Goh MH, Yeong WY, et al, 2018, Applying Macromolecular Crowding to 3D Bioprinting: Fabrication of 3D Hierarchical Porous Collagen-based Hydrogel Constructs. Biomater Sci, 6:562–74. https://doi.org/10.1039/c7bm01015j

11. Lee JM, Suen SK, Ng WL, et al., 2020, Bioprinting of Collagen: Considerations, Potentials, and Applications. Macromol Biosci, 21:2000280. https://doi.org/10.1002/mabi.202000280

12. Osidak EO, Kozhukhov VI, Osidak MS, et al., 2020, Collagen as Bioink for Bioprinting: A Comprehensive Review. Int J Bioprint, 6:270. https://doi.org/10.18063/ijb.v6i3.270

13. Ng WL, Ayi TC, Liu YC, et al., 2021, Fabrication and Characterization of 3D Bioprinted Triple-layered Human Alveolar Lung Models. Int J Bioprint, 7:332. https://doi.org/10.18063/ijb.v7i2.332

14. Lee JM, Sing SL, Tan EY, et al., 2016, Bioprinting in Cardiovascular Tissue Engineering: A Review. Int J Bioprint, 2:136–45. https://doi.org/10.18063/ijb.2016.02.006

15. Saunders RE, Derby B, 2014, Inkjet Printing Biomaterials for Tissue Engineering: Bioprinting. Int Mater Rev, 59:430–48. https://doi.org/10.1179/1743280414y.0000000040

16. Ng WL, Lee JM, Yeong WY, et al., 2017, Microvalve-based Bioprinting Process, Bio-inks and Applications. Biomater Sci, 5:632–47. https://doi.org/10.1039/c6bm00861e

17. Ng WL, Yeong WY, Naing MW, 2016, Microvalve Bioprinting of Cellular Droplets with High Resolution and Consistency. Proceedings of the International Conference on Progress in Additive Manufacturing, p397–402.

18. Koch L, Brandt O, Deiwick A, et al., 2017, Laser Assisted Bioprinting at Different Wavelengths and Pulse Durations with a Metal Dynamic Release Layer: Aa Parametric Study. Int J Bioprint, 3:42–53. https://doi.org/10.18063/ijb.2017.01.001

19. Guo F, Li P, French JB, et al., 2015, Controlling Cell-Cell Interactions Using Surface Acoustic Waves. Proc Natl Acad Sci U S A, 112:43–8. https://doi.org/10.1073/pnas.1422068112

20. Choe YE, Kim GH, 2020, A PCL/Cellulose Coil-shaped Scaffold Via a Modified Electrohydrodynamic Jetting Process. Virtual Phys Prototyp, 15:403–16. https://doi.org/10.1080/17452759.2020.1808269

21. Ozbolat IT, Hospodiuk M, 2016, Current Advances and Future Perspectives in Extrusion-Based Bioprinting. Biomaterials, 76:321–43. https://doi.org/10.1016/j.biomaterials.2015.10.076

22. Ng WL, Yeong WY, Naing MW, 2016, Development of Polyelectrolyte Chitosan-gelatin Hydrogels for Skin Bioprinting. Proc CIRP, 49:105–12. https://doi.org/10.1016/j.procir.2015.09.002

23. Ng WL, Yeong WY, Naing MW, 2016, Polyelectrolyte Gelatin-chitosan Hydrogel Optimized for 3D Bioprinting in Skin Tissue Engineering. Int J Bioprint, 2:53–62. https://doi.org/10.18063/ijb.2016.01.009

24. Zhuang P, Ng WL, An J, et al., 2019, Layer-by-layer Ultraviolet Assisted Extrusion-Based (UAE) Bioprinting of Hydrogel Constructs with High Aspect Ratio for Soft Tissue Engineering Applications. PLoS One, 14:e0216776. https://doi.org/10.1371/journal.pone.0216776

25. Ng WL, Yeong WY, Naing MW, 2014, Potential of Bioprinted Films for Skin Tissue Engineering. Proceedings of the 1st International Conference on Progress in Additive Manufacturing, p441–6. https://doi.org/10.3850/978-981-09-0446-3_065

26. Meng Z, He J, Li J, et al., 2020, Melt-based, Solvent-free Additive Manufacturing of Biodegradable Polymeric Scaffolds with Designer Microstructures for Tailored Mechanical/Biological Properties and Clinical Applications. Virtual Phys Prototyp, 15:417–44. https://doi.org/10.1080/17452759.2020.1808937

27. Ng WL, Lee JM, Zhou M, et al., 2020, Vat Polymerization based Bioprinting-process, Materials, Applications and Regulatory Challenges. Biofabrication, 12:022001. https://doi.org/10.1088/1758-5090/ab6034

28. Li W, Mille LS, Robledo JA, et al., 2020, Recent Advances in Formulating and Processing Biomaterial Inks for Vat Polymerization-Based 3D Printing. Adv Healthc Mater, 9:2000156. https://doi.org/10.1002/adhm.202000156

29. Nieto D, Corrales JA, de Mora AJ, et al., 2020, Fundamentals of Light-Cell-Polymer Interactions in Photo-cross-linking Based Bioprinting. APL Bioeng, 4:041502. https://doi.org/10.1063/5.0022693

30. Gudupati H, Dey M, Ozbolat I, 2016, A Comprehensive Review on Droplet-based Bioprinting: Past, Present and Future. Biomaterials, 102:20–42. https://doi.org/10.1016/j.biomaterials.2016.06.012

31. Ng WL, Tan ZQ, Yeong WY, et al., 2018, Proof-of-concept: 3D Bioprinting of Pigmented Human Skin Constructs. Biofabrication, 10:025005. https://doi.org/10.1088/1758-5090/aa9e1e

32. Ng WL, Wang S, Yeong WY, et al., 2016, Skin Bioprinting: Impending Reality or Fantasy? Trends Biotechnol, 34:689–99. https://doi.org/10.1016/j.tibtech.2016.04.006

33. Ng WL, Yeong WY, 2019, The Future of Skin Toxicology Testing 3D Bioprinting Meets Microfluidics. Int J Bioprint, 5:237. https://doi.org/10.18063/ijb.v5i2.1.237

34. Worthington AM, 1877, XXVIII. On the Forms Assumed by Drops of Liquids Falling Vertically on a Horizontal Plate. Proc R Soc London, 25:261–72. https://doi.org/10.1098/rspl.1876.0048

35. Thoroddsen ST, Etoh TG, Takehara K, 2008, High-Speed Imaging of Drops and Bubbles. Annu Rev Fluid Mech, 40:257–85. https://doi.org/10.1146/annurev.fluid.40.111406.102215

36. Lepowsky E, Muradoglu M, Tasoglu S, 2018, Towards Preserving Post-printing Cell Viability and Improving the Resolution: Past, Present, and Future of 3D Bioprinting Theory. Bioprinting, 11:e00034. https://doi.org/10.1016/j.bprint.2018.e00034

37. Tasoglu S, Kaynak G, Szeri AJ, et al., 2010, Impact of a Compound Droplet on a Flat Surface: A Model for Single Cell Epitaxy. Phys Fluids, 22:082103. https://doi.org/10.1063/1.3475527

38. Nooranidoost M, Izbassarov D, Tasoglu S, et al., 2019, A Computational Study of Droplet-based Bioprinting: Effects of Viscoelasticity. Phys Fluids, 31:081901. https://doi.org/10.1063/1.5108824

39. Hendriks J, Visser CW, Henke S, et al., 2015, Optimizing Cell Viability in Droplet-based Cell Deposition. Sci Rep, 5:11304. https://doi.org/10.1038/srep11304

40. Gorr HM, Zueger JM, McAdams DR, et al., 2013, Salt-induced Pattern Formation in Evaporating Droplets of Lysozyme Solutions. Colloids Surf B Biointerfaces, 103:59–66. https://doi.org/10.1016/j.colsurfb.2012.09.043

41. Rutgers IR, 1962, Relative Viscosity of Suspensions of Rigid Spheres in Newtonian liquids. Rheol Acta, 2:202–10.

42. Ng WL, Yeong WY, Naing MW, 2017, Polyvinylpyrrolidone-Based Bio-Ink Improves Cell Viability and Homogeneity during Drop-On-Demand Printing. Materials, 10:190. https://doi.org/10.3390/ma10020190

43. Xu C, Zhang M, Huang Y, et al., 2014, Study of Droplet Formation Process during Drop-on-Demand Ink jetting of Living Cell-Laden Bioink. Langmuir, 30:9130–8. https://doi.org/10.1021/la501430x

44. Dong L, Johnson D, 2003, Surface Tension of Charge stabilized Colloidal Suspensions at the Water Air Interface. Langmuir, 19:10205–9. https://doi.org/10.1021/la035128j

45. Jang D, Kim D, Moon J, 2009, Influence of Fluid Physical Properties on Ink-jet Printability. Langmuir, 25:2629–35. https://doi.org/10.1021/la900059m

46. Lee A, Sudau K, Ahn KH, et al., 2012, Optimization of Experimental Parameters to Suppress Nozzle Clogging in Inkjet Printing. Ind Eng Chem Res, 51:13195–204. https://doi.org/10.1021/ie301403g

47. Srichan C, Saikrajang T, Lomas T, et al., 2009, Inkjet Printing PEDOT: PSS Using Desktop Inkjet Printer. 2009 6th International Conference on Electrical Engineering/Electronics, Computer, Telecommunications and Information Technology, IEEE, p465–8. https://doi.org/10.1109/ecticon.2009.5137049

48. Derby B, Reis N, 2003, Inkjet Printing of Highly Loaded Particulate Suspensions. MRS Bull, 28:815–18. https://doi.org/10.1557/mrs2003.230

49. Yarin AL, 2006, Drop Impact Dynamics: Splashing, Spreading, Receding, Bouncing. Annu Rev Fluid Mech, 38:159–92. https://doi.org/10.1146/annurev.fluid.38.050304.092144

50. Rioboo R, Tropea C, Marengo M, 2001, Outcomes from a Drop Impact on Solid Surfaces. Atomization Sprays, 11:155–66. https://doi.org/10.1615/atomizspr.v11.i2.40

51. Josserand C, Thoroddsen ST, 2016, Drop Impact on a Solid Surface. Annu Rev Fluid Mech, 48:365–91. https://doi.org/10.1146/annurev-fluid-122414-034401

52. Stow CD, Hadfield MG, 1981, An Experimental Investigation of Fluid Flow Resulting from the Impact of a Water Drop with an Unyielding Dry Surface. Proc R Soc London A Math Phys Sci, 373:419–41. https://doi.org/10.1098/rspa.1981.0002

53. Mundo C, Sommerfeld M, Tropea C, 1995, Droplet-wall Collisions: Experimental Studies of the Deformation and Breakup Process. Int J Multiphase Flow, 21:151–73. https://doi.org/10.1016/0301-9322(94)00069-v

54. Yarin AL, Weiss DA, 1995, Impact of Drops on Solid Surfaces: Self-similar Capillary Waves, and Splashing as a New Type of Kinematic Discontinuity. J Fluid Mech, 283:141–73. https://doi.org/10.1017/s0022112095002266

55. Wal RL, Berger GM, Mozes SD, 2006, The Splash/Nonsplash Boundary Upon a Dry Surface and thin Fluid Film. Exp Fluids, 40:53–9. https://doi.org/10.1007/s00348-005-0045-1

56. Moreira AL, Moita AS, Panão MR, 2010, Advances and challenges in explaining fuel spray impingement: How much of single droplet impact research is useful? Prog Energy Combust Sci, 36:554–80. https://doi.org/10.1016/j.pecs.2010.01.002

57. Lee JB, Derome D, Guyer R, et al., 2016, Modeling the Maximum Spreading of Liquid Droplets Impacting Wetting and Nonwetting Surfaces. Langmuir, 32:1299–308. https://doi.org/10.1021/acs.langmuir.5b04557

58. Shaikeea AJ, Basu S, Tyagi A, et al., 2017, Universal Representations of Evaporation Modes in Sessile Droplets. PLoS One, 12:e0184997. https://doi.org/10.1371/journal.pone.0184997

59. Goh GL, Saengchairat N, Agarwala S, et al., 2019, Sessile Droplets Containing Carbon Nanotubes: A Study of Evaporation Dynamics and CNT Alignment for Printed Electronics. Nanoscale, 11:10603–14. https://doi.org/10.1039/c9nr03261d

60. Birdi K, Vu D, 1993, Wettability and the Evaporation Rates of Fluids from Solid Surfaces. J Adhes Sci Technol, 7:485–93. https://doi.org/10.1163/156856193x00808

61. Hu H, Larson RG, 2002, Evaporation of a Sessile Droplet on a Substrate. J Phys Chem B, 106:1334–44.

62. Maeno E, Takahashi N, Okada Y, Dysfunction of Regulatory Volume Increase is a Key Component of Apoptosis. FEBS Lett, 580:6513–7. https://doi.org/10.1016/j.febslet.2006.10.074

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International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing