Laser-induced Forward Transfer Hydrogel Printing: A Defined Route for Highly Controlled Process

Vladimir Yusupov, Semyon Churbanov, Ekaterina Churbanova, Ksenia Bardakova, Artem Antoshin, Stanislav Evlashin, Peter Timashev, Nikita Minaev

Article ID: 271
Vol 6, Issue 3, 2020, Article identifier:271

VIEWS - 611 (Abstract) 155 (PDF)


Laser-induced forward transfer is a versatile, non-contact, and nozzle-free printing technique which has demonstrated high potential for different printing applications with high resolution. In this article, three most widely used hydrogels in bioprinting (2% hyaluronic acid sodium salt, 1% methylcellulose, and 1% sodium alginate) were used to study laser printing processes. For this purpose, the authors applied a laser system based on a pulsed infrared laser (1064 nm wavelength, 8 ns pulse duration, 1 – 5 J/cm2 laser fluence, and 30 μm laser spot size). A high-speed shooting showed that the increase in fluence caused a sequential change in the transfer regimes: No transfer regime, optimal jetting regime with a single droplet transfer, high speed regime, turbulent regime, and plume regime. It was demonstrated that in the optimal jetting regime, which led to printing with single droplets, the size and volume of droplets transferred to the acceptor slide increased almost linearly with the increase of laser fluence. It was also shown that the maintenance of a stable temperature (±2°C) allowed for neglecting the temperature-induced viscosity change of hydrogels. It was determined that under room conditions (20°C, humidity 50%), the hydrogel layer, due to drying processes, decreased with a speed of about 8 μm/min, which could lead to a temporal variation of the transfer process parameters. The authors developed a practical algorithm that allowed quick configuration of the laser printing process on an applied experimental setup. The configuration is provided by the change of the easily tunable parameters: Laser pulse energy, laser spot size, the distance between the donor ribbon and acceptor plate, as well as the thickness of the hydrogel layer on the donor ribbon slide.


LIFT, Laser-induced forward transfer, Hydrogel parameters, Optimal jetting regime, Jet and droplets parameters

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Serra P, Fernández-Pradas JM, Berthet FX, et al., 2004, Laser Direct Writing of Biomolecule Microarrays. Appl Phys A Mater Sci Process, 79:949–952. DOI: 10.1007/s00339-004-2577-2.

Zergioti I, Karaiskou A, Papazoglou DG, et al., 2005, Femtosecond Laser Microprinting of Biomaterials. Appl Phys Lett, 86:1–3. DOI: 10.1063/1.1906325.

Colina M, Serra P, Fernández-Pradas JM, et al., 2005, DNA Deposition through Laser Induced Forward Transfer. Biosens. Bioelectron., 20(8):1638–1642. DOI: 10.1016/j.bios.2004.08.047.

Serra P, Colina M, Fernández-Pradas JM, et al., 2004, Preparation of Functional DNA Microarrays through Laser induced Forward Transfer. Appl Phys Lett, 85:1639–1641.DOI: 10.1063/1.1787614.

Cheptsov VS, Tsypina SI, Minaev NV, et al., 2019, New Microorganism Isolation Techniques with Emphasis on Laser Printing. Int J Bioprinting, 5:1–12. DOI: 10.18063/ijb.v5i1.165.

Koch L, Deiwick A, Schlie S, et al., 2012, Skin Tissue Generation by Laser Cell Printing. Biotechnol. Bioeng.,109:1855–1863. DOI: 10.1002/bit.24455.

Gaebel R, Liu J, Koch, L, et al., 2011, Patterning Human Stem Cells and Endothelial Cells with Laser Printing for Cardiac Regeneration. Biomaterials, 32:9218–9230. DOI:10.1016/j.biomaterials.2011.08.071.

Gaebel R, Kuhn S, Sorg H, et al., 2009, Laser printing of skin cells and human stem cells. Tissue Eng Part C Methods,16:847–854. DOI: 10.1089/ten.tec.2009.0397.

Mandrycky C, Wang Z, Kim K, et al., 2016, 3D Bioprinting for Engineering Complex Tissues. Biotechnol. Adv, 34:422–434. DOI: 10.1016/j.biotechadv.2015.12.011.

Koch L, Kuhn S, Sorg H, et al., 2010, Laser Printing of Skin Cells and Human Stem Cells. Tissue Eng Part C Methods,16:847–854. DOI: 10.1089/ten.tec.2009.0397.

Catros S, Fricain JC, Guillotin B, et al., 2011, Laser-assisted Bioprinting for Creating on-demand Patterns of Human Osteoprogenitor Cells and Nano-hydroxyapatite. Biofabrication, 3:025001. DOI: 10.1088/1758-5082/3/2/025001.

Hopp B, Smausz T, Antal Z, et al., 2004, Absorbing Film Assisted Laser Induced Forward Transfer of Fungi (Trichoderma conidia). J Appl Phys, 96:3478–3481. DOI:10.1063/1.1782275.

Yusupov VI, Zhigarkov VS, Churbanova ES, et al., 2017, Laser-induced Transfer of Gel Microdroplets for Cell Printing. Quantum Electron, 47:1158–1165. DOI: 10.1070/qel16512.

Zhang Z, Xiong R, Mei R, et al,. 2015, Time-Resolved Imaging Study of Jetting Dynamics during Laser Printing of Viscoelastic Alginate Solutions. Langmuir, 31:6447–6456. DOI: 10.1021/acs.langmuir.5b00919.

Zhang Z, Xiong R, Corr DT, et al., 2016, Study of Impingement Types and Printing Quality during Laser Printing of Viscoelastic Alginate Solutions. Langmuir, 32:3004–3014. DOI: 10.1021/acs.langmuir.6b00220.

Guillotin B, Souquet A, Catros S, et al., 2010, Laser Assisted Bioprinting of Engineered Tissue with High Cell Density and Microscale Organization. Biomaterials, 31:7250–7256. DOI: 10.1016/j.biomaterials.2010.05.055.

Gaebel R, Ma N, Liu J, et al., 2011, Patterning Human Stem Cells and Endothelial Cells with Laser Printing for Cardiac Regeneration. Biomaterials, 32:9218–9230. DOI: 10.1016/j. biomaterials.2011.08.071.

Gruene M, Pflaum M, Hess C, et al., 2011, Laser Printing of Three-dimensional Multicellular Arrays for Studies of Cellcell and Cell-environment Interactions. Tissue Eng Part C Methods, 17:973–982. DOI: 10.1089/ten.tec.2011.0185.

Koch L, Brandt O, Deiwick A, et al., 2017, Laser-assisted Bioprinting at Different Wavelengths and Pulse Durations with a Metal Dynamic Release Layer: A Parametric Study. Int J Bioprinting, 3:42–53. DOI: 10.18063/ijb.2017.01.001.

Sorkio A, Koch L, Koivusalo L, et al., 2018, Human Stem Cell Based Corneal Tissue Mimicking Structures Using Laser assisted 3D Bioprinting and Functional Bioinks. Biomaterials, 171:57–71. DOI: 10.1016/j.biomaterials.2018.04.034.

Guillemot F, Souquet A, Catros S, et al., 2010, High throughput Laser Printing of Cells and Biomaterials for Tissue Engineering. Acta Biomater, 6:2494–2500. DOI:10.1016/j.actbio.2009.09.029.

Catros S, Guillemot F, Nandakumar A, et al., 2011, Layer by-Layer Tissue Microfabrication Supports Cell Proliferation In Vitro and In Vivo. Tissue Eng Part C Methods, 18:62–70. DOI: 10.1089/ten.tec.2011.0382.

Michael S, Sorg H, Peck CT, et al., 2013, Tissue Engineered Skin Substitutes Created by Laser-Assisted Bioprinting form Skin-Like Structures in the Dorsal Skin Fold Chamber in Mice. PLoS One, 8:e57741. DOI: 10.1371/journal.pone.0057741.

Antoshin AA, Churbanov SN, Minaev NV, et al., 2019, LIFT Bioprinting, is it Worth it? Bioprinting, 15:e00052. DOI:10.1016/j.bprint.2019.e00052.

Bashkatov AN, Genina EA, Kochubey VI, et al., 2005, Optical Properties of Human Skin Subcutaneous and Mucous Tissues in the Wavelength Range from 400 to 2000 nm. J Phys D Appl Physics, 38:2543–2555. DOI: 10.1088/0022-3727/38/15/004.

Carvalho S, Gueiral N, Nogueira E, et al., 2017, Comparative Study of the Optical Properties of Colon Mucosa and Colon Precancerous Polyps between 400 and 1000 nm. Dynamics and Fluctuations in Biomedical Photonics XIVXIV International Society for Optics and Photonics, 10063:100631L. DOI:10.1117/12.2253023.

Pagès E, Rémy M, Kériquel V, et al., 2015, Creation of Highly Defined Mesenchymal Stem Cell Patterns in Three Dimensions by Laser-Assisted Bioprinting. J Nanotechnol Eng Med, 6:021005. DOI: 10.1115/1.4031217.

Zhang Z, Xu C, Xiong R, et al., 2017, Effects of Living Cells on the Bioink Printability during Laser Printing. Biomicrofluidics, 11:034120. DOI: 10.1063/1.4985652.

Zhang Z, Chai W, Xiong R, et al., 2017, Printing-induced Cell Injury Evaluation during Laser Printing of 3T3 Mouse Fibroblasts. Biofabrication, 9:025038. DOI: 10.1088/1758-5090/aa6ed9.

Unger C, Gruene M, Koch L, et al., 2011, Time-resolved Imaging of Hydrogel Printing Via Laser-induced Forward Transfer. Appl Phys A Mater Sci Process, 103:271–277. DOI:10.1007/s00339-010-6030-4.

Desrus H, Chassagne B, Catros S, et al., 2016, Laser Assisted

Bioprinting using a Femtosecond Laser with and without a Gold Transductive Layer: A Parametric Study. Proceedings Volume 9706, Optical Interactions with Tissue and Cells XXVII. DOI: 10.1117/12.2209087.

Cheptsov VS, Churbanova ES, Yusupov VI, et al., 2018, Laser Printing of Microbial Systems: Effect of Absorbing Metal Film. Lett Appl Microbiol, 67:544–549. DOI: 10.1111/lam.13074.

Riester D, Budde J, Gach C, et al., 2016, High Speed Photography of Laser Induced Forward Transfer (LIFT) of Single and Double-layered Transfer Layers for Single Cell Transfer. J Laser Micro Nanoeng, 11:199–203. DOI: 10.2961/jlmn.2016.02.0010.

Zarubin VP, Zhigarkov VS, Yusupov VI, et al., 2019, Physical Processes Affecting the Survival of Microbiological Systems in Laser Printing of Gel Droplets. Quantum Electron,49:1068–1073. DOI: 10.1070/qel17081.

Tomasina S, Bodet C, Mota T, et al., 2019, Bioprinting Vasculature: Materials, Cells and Emergent Techniques.Materials (Basel), 12:2701. DOI: 10.3390/ma12172701.

Young HD, Modi R, Bucaro M, 2002, Generation of Mesoscopic Patterns of Viable Escherichia coli by Ambient Laser Transfer. Biomaterials, 23:161–166. DOI: 10.1016/s0142-9612(01)00091-6.

Xiong R, Zhang Z, Huang Y, 2015, Identification of Optimal Printing Conditions for Laser Printing of Alginate Tubular Constructs. J Manuf Process, 20:450–455. DOI: 10.1016/j.jmapro.2015.06.023.

Palla-Papavlu A, Córdoba C, Patrascioiu A, et al., 2013, Deposition and Characterization of Lines Printed through Laser-induced Forward Transfer. Appl Phys A Mater Sci Process, 110:751–755. DOI: 10.1007/s00339-012-7279-6.

Pescosolido L, Miatto S, Di Meo C, et al., 2010, Injectable and In Situ Gelling Hydrogels for Modified Protein Release. Eur Biophys J, 39:903-9. DOI: 10.1007/s00249-009-0440-2.

Ouyang L, Highley CB, Rodell CB, et al., 2016, 3D Printing of Shear-Thinning Hyaluronic Acid Hydrogels with Secondary Cross-Linking. ACS Biomater Sci Eng, 2:1743–1751. DOI: 10.1021/acsbiomaterials.6b00158.

Cochis A, Bonetti L, Sorrentino R, et al., 2018, 3D Printing of Thermo-responsive Methylcellulose Hydrogels for Cell-sheet Engineering. Materials (Basel), 11:1–14. DOI: 10.3390/ma11040579.

Ovsianikov A, Gruene M, Pflaum M, et al., 2010, Laser Printing of Cells into 3D Scaffolds. Biofabrication, 2:014104. DOI: 10.1088/1758-5082/2/1/014104.

Gruene M, Pflaum M, Deiwick A, et al., 2011, Adipogenic Differentiation of Laser-printed 3D Tissue Grafts Consisting of Human Adipose-derived Stem Cells. Biofabrication, 3:015005. DOI: 10.1088/1758-5082/3/1/015005.

Yusupov VI, Gorlenko MV, Cheptsov VS, et al., 2018, Laser Engineering of Microbial Systems. Laser Phys Lett, 15:015604.

Unger C, Koch J, Overmeyer L, et al., 2012, Time-resolved Studies of Femtosecond-laser Induced Melt Dynamics. Opt Express, 20:24864. DOI: 10.1364/oe.20.024864.

Hill C, 2016, Liquid-Phase Laser Induced Forward Transfer for Complex Organic Inks and Tissue Engineering. Ann Biomed Eng, 45:84–99. DOI: 10.1007/s10439-016-1617-3.

Akhatov W, Lindau I, Topolnikov O, et al., 2001, Collapse and Rebound of a Laser-induced Cavitation Bubble. Phys Fluids, 13:29–32. DOI: 10.1063/1.1401810.

Mézel C, Hallo L, Souquet A, et al., 2009, Self-consistent Modeling of Jet Formation Process in the Nanosecond Laser Pulse Regime. Phys Plasmas, 16:123112. DOI:10.1063/1.3276101.

Ali M, Pages E, Ducom A, et al., 2014, Controlling Laser induced Jet Formation for Bioprinting Mesenchymal Stem Cells with High Viability and High Resolution. Biofabrication, 6:045001. DOI: 10.1088/1758-5082/6/4/045001.

Wang W, Chrisey DB, 2008, Study of Impact-Induced Mechanical Effects in Cell Direct. J Manuf Sci Eng, 130:1–10.

Ringeisen BR, Pages E, Ducom A, et al., 2004, Laser Printing of Pluripotent Embryonal Carcinoma Cells. Tissue Eng,10:483–491.

Kawecki F, Clafshenkel WP, Auger FA, et al., 2018, Self-assembled Human Osseous Cell Sheets as Living Biopapers for the Laser-assisted Bioprinting of Human Endothelial Cells. Biofabrication, 10:035006. DOI: 10.1088/1758-5090/aabd5b.

Vass C, Smausz T, Hopp B, 2004, Wet Etching of Fused Silica: A Multiplex Study. J Phys D Appl Phys, 37:2449–2454. DOI: 10.1088/0022-3727/37/17/018.

Gruene M, Unger C, Koch L, et al., 2011, Dispensing Pico to Nanolitre of a Natural Hydrogel by Laser assisted Bioprinting. Biomed Eng Online, 10:9–12. DOI:10.1186/1475-925x-10-19.

Brown MS, Brasz CF, Ventikos Y, Arnold CB, 2012, Impulsively Actuated Jets from thin Liquid Films for High resolution Printing Applications. J Fluid Mech, 709:341–370. DOI: 10.1017/jfm.2012.337.

Hölzl K, Lin S, Tytgat L, et al., 2016, Bioink Properties before, during and after 3D Bioprinting. Biofabrication, 8:032002. DOI: 10.1088/1758-5090/8/3/032002.

Ringeisen BR, Spargo BJ, Wu PK, 2010, Cell and Organ Printing. Springer Science and Business Media, Berlin.

Nasatto PL, Pignon F, Silveira JL, et al., 2015, Methylcellulose, a Cellulose Derivative with Original Physical Properties and Extended Applications. Polymers (Basel), 7:777–803. DOI: 10.3390/polym7050777.

Shafiee A, Elham G, Ramesh H, et al., 2019, Physics of Bioprinting. Appl Phys Rev, 6:021315.

McKinley GH, Renardy M, 2011, Wolfgang von Ohnesorge. Phys Fluids, 23:127101. DOI: 10.1063/1.3663616.

Yan J, Huang Y, Xu C, et al., 2012, Effects of Fluid Properties and Laser Fluence on Jet Formation during Laser Direct Writing of Glycerol Solution. J Appl Phys, 112:083105. DOI:10.1063/1.4759344.

Bush JW, 2004, Rayleigh Instability. In: MIT Lecture Notes on Surface Tension, Lecture 5 (PDF) Massachusetts Institute of Technology.

Gorlenko M, Chutko EA, Churbanova ES, et al., 2018, Laser Microsampling of Soil Microbial Community. J Biol Eng, 12:1–11.

Minaev NV, Chutko EA, Churbanova ES, et al., 2018, Laser Printing of Gel Microdrops with Living Cells and Microorganisms. KnE Energy Phys, 2018:23–31. DOI:10.18502/ken.v3i3.2010.

Koch L, Deiwick A, Franke A, et al., 2018, Laser Bioprinting of Human Induced Pluripotent Stem Cells the Effect of Printing and Biomaterials on Cell Survival, Pluripotency, and Differentiation. Biofabrication, 10:035005. DOI:10.1088/1758-5090/aab981.

Cheptsov VS, Tsypina SI, Minaev NV, et al., 2019, New Microorganism Isolation Techniques With Emphasis On Laser Printing. Int J Bioprint, 5:165. DOI:10.18063/ijb.v5i1.165.



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