Laser Biofabrication

Research Focus

The aim of this unit is to develop intelligent biomedical implants, 3D scaffolds for tissue engineering, novel photosensitive bioactive and biodegradable materials, and artifical tissue replacements and organs for regenerative medicine. This involves investigations into the influence of 2D and 3D microstructures on cell functionalities, demonstration of technological possibilities for 3D arrangement of different cells by laser printing, and exploration of the scope for fabricating vascularized tissue.

By using two-photon polymerization (2PP), scaffolds can be generated by directly writing CAD structures into the volume of polymer solutions. The polymerization occurs in the laser focus only. Thereby, resolutions below the diffraction limit down to the sub-100-nanometer range have been achieved. Scaffolds from different biomaterials, including organic-inorganic sol-gel-composites, biodegradable polymers, and hydrogels or blends, have been generated with this technique. By changing the topological and biochemical characteristics of the nanostructured surfaces, control over cell proliferation and cell interaction can be provided. The ultimate aim is to mimic and reproduce natural ECM (extra cellular matrix) environment architecture and properties with artificially created micro- and nanostructures.
Within the REBIRTH Cluster of Excellence, fundamental aspects of the way femtosecond laser radiation interacts with the applied materials are explored, as are practical aspects of nano-manufacturing. Resolution limits and biological properties of fabricated structures are investigated. Producing series of exactly identical samples in accordance with CAD models will allow systematic studies of cell-structure interactions in 3D and new insights into tissue formation mechanisms.

Biological Laser Printing as a further technique provides unique possibilities for the arrangement of different living cells, micro-organisms and biomaterials in well-defined 2D and 3D patterns. Focused laser pulses are applied to transfer tiny droplets (with volumes in the picoliter range) of biocompatible hydrogels with embedded living cells in pre-defined positions. Layer-by-layer 3D tissue constructs are generated. It was demonstrated that cells are not harmed by the printing process and the differentiation behavior of stem cells is not affected. However, biomaterials need to be adapted to the printed cells and the intended application.

This printing technique was applied for generation of skin tissue from dermal fibroblasts and epidermal keratinocytes. It was demonstrated that the cells formed intercellular adherens junctions and functional gap junctions, which is seen as a proof for tissue formation. Implanting the printed skin tissue in full thickness wounds in mice shows good ingrowth of the printed skin in the native mouse skin. Besides the generation of tissue, this technique was also used to arrange cells in specific patterns to study cell-cell and cell-environment interactions. Cell behavior in conventional 2D cell culture differs considerably from the behavior in vivo or in 3D tissue structures. In contrast, 3D cell printing aims to arrange and study cells in their specific in-vivo-like micro-environment.

Both techniques are capable of advancing 3D cell culture towards CAD defined and precisely arranged 3D cell models. Such innovative 3D cell models could provide new insights in understanding of cell behavior, tissue functions and regeneration. Printed tissue, for example skin, can be used for analyzing the effect of agents like pharmaceuticals or cosmetics ex vivo and, by applying human primary cells, might be more relevant than animal tests.



  • Prof. A. Haverich, M. Pflaum, Hannover Medical School, LEBAO, REBIRTH Unit Biohybrid Lung, Printing different cell types in arrays for investigating cell-cell interactions + Transfer of endothelial cells on different supporting structures to form artificial vessels
  • Dr. A. Schambach, Hannover Medical School, REBIRTH Unit Regenerative Gene Therapy, Printing of transfected cells for long-term investigations of cell migration
  • Dr. H. Meyer, Laser Zentrum Hannover e.V., REBIRTH Unit Laser Manipulation and Cellular Engineering, Dr. A. Loos, BioMedimplant, REBIRTH Unit Biocompatibility, Prof. E. Ponimaskin, Hannover Medical School, Visualization of printed 3D cell structures
  • Dr. F. Limbourg, Hannover Medical School, REBIRTH Unit Regenerative Agents, Printing vascular tubes for analysis of signaling pathways and tubulogenesis
  • Dr. R. Zweigerdt, REBIRTH Unit Mass Production of Pluripotent Stem Cells, Leibniz Research Laboratories for Biotechnology and Artificial Organs, Hannover Medical School
  • Prof. M. Schiffer, Hannover Medical School, Nephrology, Printing of Podocytes, mesangial and endothelial cells to mimick the Kidney glomerulus
  • Prof. B. Taidi, Laboratoire Génie des Procédés et Matériaux, École Centrale Paris, France, Printing of micro-organisms in specific patterns to analyze interactions between different species


  • University of Rostock, Institut für Biomedizinische Technik (IBMT)
  • Hannover Medical School, Institute for Molecular- and Cellphysiology
  • Leibniz University of Hannover, Institute of Technical Chemistry
  • RWTH Aachen
  • Leibniz University of Hannover, Institute of Biophysics
  • North Carolina State University, USA
  • Sechenov University, Institute for Regenerative Medicine, Moscow, Russia
  • Lobachevsky State University of Nizhny Novgorod, Nizhny Novgorod, Russia
  • G. Draeger, REBIRTH Unit Functionalized Polymers and Regenerative Agents, Leibniz University of Hannover: Novel biodegradable photosensitive materials for 2PP

Further Research Projects

  • Biofabrication for NIFE, Niedersächsisches Vorab, Volkswagen Stiftung
  • ‘Sub-100 nm Zweiphotonen- Polymerisations-Technik für optische und biomedizinische Anwendungen’ (in the framework of DFG SPP 1327 ‘Optisch Erzeugte Sub-100nm- Strukturen für Biomedizinische und technische Applikationen’)
  • ‘Mikrostrukturierte Cochlea- Implantat-Elektroden’ (SFB-599 ‘Zukunftsfähige bioresorbierbare und permanente Implantate aus metallischen und keramischen Werkstoffen’ – Teilprojekt T1)
  • ‘Nerven-Elektroden-Interaktion’ (SFB-599 ‘Zukunftsfähige bioresorbierbare und permanente Implantate aus metallischen und keramischen Werkstoffen’ – Teilprojekt D2)
  • ‘Micro-fabrication of polymeric lab-on-a-chip by ultrafast lasers with integrated optical detection’ (EUProject ‘Microfluidic’)
  • ‘High speed to photon polymerisation’ (EU-Project ‘2PP-Lightwave’ (Era-Spot)
  • Elektrodenoptimierung für Neuroprothesen
  • MESO-BRAIN: Custom architecturally defined 3D stem cell derived functional human neural networks for transformative progress in neuroscience and medicine. (EU project H2020-FET OPEN)
  • PrintUVSkin - Laser Printed Skin with natural UV protection for Implantation and Wound Dressing (B.Braun Melsungen AG)


  • B. Chichkov, Award of the HannoverImpuls GmbH for the following innovation: ‘M3D - Verfahren und Maschine zur 3D-Mikrostrukturierung’, (2007)


  • Lecture ‘Photonics’ at Leibniz University Hannover, Prof. B. Chichkov


2013 - ongoing


Kuznetsova D, Ageykin A, Koroleva A, Deiwick A, Shpichka A, Solovieva A, Kostjuk S, Meleshina A, Rodimova S, Akovanceva A, Butnaru D, Frolova A, Zagaynova E, Chichkov B, Bagratashvili V, Timashev P. Surface micromorphology of cross-linked tetrafunctional polylactide scaffolds inducing vessel growth and bone formation. Biofabrication. 2017;9(2):025009.

Shpichka AI, Koroleva AV, Deiwick A, Timashev PS, Semenova EF, Moiseeva IY, Konoplyannikov MA, Chichkov BN. Evaluation of the vasculogenic potential of hydrogels based on modified fibrin. Cell and Tissue Biology. 2017;11(1):81-7.

Koch L, Deiwick A, Chichkov B. 16 - Laser additive printing of cells A2 - Brandt, Milan.  Laser Additive Manufacturing: Woodhead Publishing; 2017. p. 421-37.

Koch L, Brandt O, Deiwick A, Chichkov B. Laser assisted bioprinting at different wavelengths and pulse durations with a metal dynamic release layer: A parametric study. 2017. 2017;3(1).


Roger Y, Schack LM, Koroleva A, Noack S, Kurselis K, Krettek C, Chichkov B, Lenarz T, Warnecke A, Hoffmann A. Grid-like surface structures in thermoplastic polyurethane induce anti-inflammatory and anti-fibrotic processes in bone marrow-derived mesenchymal stem cells. Colloids and Surfaces B-Biointerfaces. 2016;148:104-15.

Taidi B, Lebernede G, Koch L, Perre P, Chichkov B. Colony development of laser printed eukaryotic (yeast and microalga) microorganisms in co-culture. 2016. 2016;2(2).

Koroleva A, Deiwick A, Nguyen A, Narayan R, Shpichka A, Kufelt O, Kiyan R, Bagratashvili V, Timashev P, Scheper T, Chichkov B. Hydrogel-based microfluidics for vascular tissue engineering. BioNanoMaterials2016. p. 19.

Koch L, Deiwick A, Chichkov B. Laser-Based Cell Printing. In: Ovsianikov A, Yoo J, Mironov V, editors. 3D Printing and Biofabrication. Cham: Springer International Publishing; 2016. p. 1-27.


Kufelt O, El-Tamer A, Sehring C, Meissner M, Schlie-Wolter S, Chichkov BN. Water-soluble photopolymerizable chitosan hydrogels for biofabrication via two-photon polymerization. Acta biomaterialia. 2015;18:186-95.

Chichkov B, Zywiety U, Koch L. Laser printing of silicon nanoparticles and living cells. SPIE Newsroom. 03/2015. DOI: 10.1117/2.1201503.005750

Timashev PS, Demina TS, Minaev NV, Bardakova KN, Koroleva AV, Kufelt OA, Chichkov BN, Panchenko VY, Akopova TA, Bagratashvili VN. Fabrication of Microstructured Materials Based on Chitosan and Its Derivatives Using Two-Photon Polymerization. High Energy Chemistry. 2015;49(4):300-3.

Kufelt O, El-Tamer A, Sehring C, Meissner M, Schlie-Wolter S, Chichkov BN. Water-Soluble Photopolymerizable Chitosan Hydrogels for Biofabrication Via Two-Photon Polymerization. Acta Biomater. 2015;18:186-95.

Koroleva A, Deiwick A, Nguyen A, Schlie-Wolter S, Narayan R, Timashev P, Popov V, Bagratashvili V, Chichkov B. Osteogenic Differentiation of Human Mesenchymal Stem Cells in 3-D Zr-Si Organic-Inorganic Scaffolds Produced by Two-Photon Polymerization Technique. PLoS One. 2015;10(2):e0118164.

Koch L, Michael S, Reimers K, Vogt PM, Chichkov B. Chapter 13 - Bioprinting for Skin. In: Zhang LG, Fisher JP, Leong KW, editors. 3d Bioprinting and Nanotechnology in Tissue Engineering and Regenerative Medicine: Academic Press; 2015. p. 281-306.


Zywietz U, Reinhardt C, Evlyukhin A, Birr T, Chichkov B. Generation and Patterning of Si Nanoparticles by Femtosecond Laser Pulses. Applied Physics A. 2014;114(1):45-50.

Zywietz U, Evlyukhin AB, Reinhardt C, Chichkov BN. Laser Printing of Silicon Nanoparticles with Resonant Optical Electric and Magnetic Responses. Nat Commun. 2014;5:3402.

Popov VK, Komlev VS, Chichkov BN. Calcium Phosphate Blossom for Bone Tissue Engineering. Materials Today. 2014;17(2):96-7.

Evlyukhin AB, Eriksen RL, Cheng W, Beermann J, Reinhardt C, Petrov A, Prorok S, Eich M, Chichkov BN, Bozhevolnyi SI. Optical Spectroscopy of Single Si Nanocylinders with Magnetic and Electric Resonances. Sci Rep. 2014;4:4126.


Obata K, El-Tamer A, Koch L, Hinze U, Chichkov BN. High-Aspect 3d Two-Photon Polymerization Structuring with Widened Objective Working Range (Wow-2pp). Light Sci Appl. 2013;2:e116.

Obata K, Chichkov BN. Advanced Femtosecond Laser Micro/Nanostructuring Using Phase Modulation Technique. International Journal of Optomechatronics. 2013;7(4):296-303.

Kurselis K, Kiyan R, Bagratashvili VN, Popov VK, Chichkov BN. 3d Fabrication of All-Polymer Conductive Microstructures by Two Photon Polymerization. Opt Express. 2013;21(25):31029-35.

Kiyan Y, Kurselis K, Kiyan R, Haller H, Chichkov BN, Dumler I. Urokinase Receptor Counteracts Vascular Smooth Muscle Cell Functional Changes Induced by Surface Topography. Theranostics. 2013;3(7):516-26.

Gittard SD, Koroleva A, Nguyen AK, Fadeeva E, Gaidukeviciute A, Schlie-Wolter S, Narayan RJ, Chichkov B. Two-Photon Polymerization Microstructuring in Regenerative Medicine. Front Biosci (Elite Ed). 2013;5:602-9.

De Marco C, Gaidukeviciute A, Kiyan R, Eaton SM, Levi M, Osellame R, Chichkov BN, Turri S. A New Perfluoropolyether-Based Hydrophobic and Chemically Resistant Photoresist Structured by Two-Photon Polymerization. Langmuir. 2013;29(1):426-31.

Allemann R, Stachs O, Falke K, Schmidt W, Siewert S, Sternberg K, Chichkov B, Wree A, Schmitz KP, Guthoff RF. [New Concepts for Pressure-Controlled Glaucoma Implants]. Ophthalmologe. 2013;110(8):733-9. Neue Konzepte fur druckgesteuerte Glaukomimplantate.

2006 - 2012


Schlie S, Gruene M, Dittmar H, Chichkov BN. Dynamics of Cell Attachment: Adhesion Time and Force. Tissue Eng Part C Methods. 2012;18(9):688-96.

Schlie S, Fadeeva E, Koroleva A, Chichkov BN. Laser-Engineered Topography: Correlation between Structure Dimensions and Cell Control. J Mater Sci Mater Med. 2012;23(11):2813-9.

Osipov V, Doskolovich L, Bezus E, Cheng W, Gaidukeviciute A, Chichkov B. Fabrication of Three-Focal Diffractive Lenses by Two-Photon Polymerization Technique. Applied Physics A. 2012;107(3):525-9.

Kuršelis K, Kiyan R, Chichkov BN. Formation of Corrugated and Porous Steel Surfaces by Femtosecond Laser Irradiation. Applied Surface Science. 2012;258(22):8845-52.

Koroleva A, Gill AA, Ortega I, Haycock JW, Schlie S, Gittard SD, Chichkov BN, Claeyssens F. Two-Photon Polymerization-Generated and Micromolding-Replicated 3d Scaffolds for Peripheral Neural Tissue Engineering Applications. Biofabrication. 2012;4(2):025005.

Koch L, Deiwick A, Schlie S, Michael S, Gruene M, Coger V, Zychlinski D, Schambach A, Reimers K, Vogt PM, Chichkov B. Skin Tissue Generation by Laser Cell Printing. Biotechnol Bioeng. 2012;109(7):1855-63.

Klopsch C, Gabel R, Kaminski A, Mark P, Wang W, Toelk A, Delyagina E, Kleiner G, Koch L, Chichkov B, Mela P, Jockenhoevel S, Ma N, Steinhoff G. Spray- and Laser-Assisted Biomaterial Processing for Fast and Efficient Autologous Cell-Plus-Matrix Tissue Engineering. J Tissue Eng Regen Med. 2012. Epub 2012/12/05.

Evlyukhin AB, Novikov SM, Zywietz U, Eriksen RL, Reinhardt C, Bozhevolnyi SI, Chichkov BN. Demonstration of Magnetic Dipole Resonances of Dielectric Nanospheres in the Visible Region. Nano Lett. 2012;12(7):3749-55.

Emons M, Obata K, Binhammer T, Ovsianikov A, Chichkov BN, Morgner U. Two-Photon Polymerization Technique with Sub-50 Nm Resolution by Sub-10 Fs Laser Pulses. Optical Materials Express. 2012;2(7):942-7.

Boehm RD, Chen B, Gittard SD, Chichkov BN, Monteiro-Riviere NA, Nasir A, Narayan RJ. Two-Photon Polymerization/Micromolding of Microscale Barbs for Medical Applications. Journal of Adhesion Science and Technology. 2012;28(3-4):387-98.

Siewert S, Schultze C, Schmidt W, Hinze U, Chichkov B, Wree A, Sternberg K, Allemann R, Guthoff R, Schmitz KP. Development of a Micro-Mechanical Valve in a Novel Glaucoma Implant. Biomed Microdevices. 2012;14(5):907-20.


Schlie S, Fadeeva E, Koroleva A, Ovsianikov A, Koch J, Ngezahayo A, Chichkov BN. Laser-Based Nanoengineering of Surface Topographies for Biomedical Applications. Photonics and Nanostructures - Fundamentals and Applications. 2011;9(2):159-62.

Ovsianikov A, Deiwick A, Van Vlierberghe S, Pflaum M, Wilhelmi M, Dubruel P, Chichkov B. Laser Fabrication of 3d Gelatin Scaffolds for the Generation of Bioartificial Tissues. Materials. 2011;4(1):288.

Ovsianikov A, Deiwick A, Van Vlierberghe S, Dubruel P, Moller L, Drager G, Chichkov B. Laser Fabrication of Three-Dimensional Cad Scaffolds from Photosensitive Gelatin for Applications in Tissue Engineering. Biomacromolecules. 2011;12(4):851-8.

Kuznetsov AI, Evlyukhin AB, Goncalves MR, Reinhardt C, Koroleva A, Arnedillo ML, Kiyan R, Marti O, Chichkov BN. Laser Fabrication of Large-Scale Nanoparticle Arrays for Sensing Applications. ACS Nano. 2011;5(6):4843-9.

Suriano R, Kuznetsov A, Eaton SM, Kiyan R, Cerullo G, Osellame R, Chichkov BN, Levi M, Turri S. Femtosecond Laser Ablation of Polymeric Substrates for the Fabrication of Microfluidic Channels. Applied Surface Science. 2011;257(14):6243-50.


Obata K, Koch J, Hinze U, Chichkov BN. Multi-Focus Two-Photon Polymerization Technique Based on Individually Controlled Phase Modulation. Opt Express. 2010;18(16):17193-200.

Narayan RJ, Doraiswamy A, Chrisey DB, Chichkov BN. Medical Prototyping Using Two Photon Polymerization. Materials Today. 2010;13(12):42-8.


Heinroth F, Bremer I, Münzer S, Behrens P, Reinhardt C, Passinger S, Ohrt C, Chichkov B. Microstructured Templates Produced Using Femtosecond Laser Pulses as Templates for the Deposition of Mesoporous Silicas. Microporous and Mesoporous Materials. 2009;119(1–3):104-8.

Gittard SD, Ovsianikov A, Monteiro-Riviere NA, Lusk J, Morel P, Minghetti P, Lenardi C, Chichkov BN, Narayan RJ. Fabrication of Polymer Microneedles Using a Two-Photon Polymerization and Micromolding Process. J Diabetes Sci Technol. 2009;3(2):304-11.

Farsari M, Chichkov BN. Materials Processing: Two-Photon Fabrication. Nat Photon. 2009;3(8):450-2.

Kuznetsov AI, Koch J, Chichkov BN. Nanostructuring of Thin Gold Films by Femtosecond Lasers. Applied Physics A. 2009;94(2):221-30.


Hartmann N, Franzka S, Koch J, Ostendorf A, Chichkov BN. Subwavelength Patterning of Alkylsiloxane Monolayers Via Nonlinear Processing with Single Femtosecond Laser Pulses. Applied Physics Letters. 2008;92(22):223111.

Dinca V, Kasotakis E, Catherine J, Mourka A, Ranella A, Ovsianikov A, Chichkov BN, Farsari M, Mitraki A, Fotakis C. Directed Three-Dimensional Patterning of Self-Assembled Peptide Fibrils. Nano Lett. 2008;8(2):538-43.


Schlie S, Ngezahayo A, Ovsianikov A, Fabian T, Kolb HA, Haferkamp H, Chichkov BN. Three-Dimensional Cell Growth on Structures Fabricated from Ormocer by Two-Photon Polymerization Technique. J Biomater Appl. 2007;22(3):275-87.

Ovsianikov A, Schlie S, Ngezahayo A, Haverich A, Chichkov BN. Two-Photon Polymerization Technique for Microfabrication of Cad-Designed 3d Scaffolds from Commercially Available Photosensitive Materials. J Tissue Eng Regen Med. 2007;1(6):443-9.

Ovsianikov A, Ostendorf A, Chichkov BN. Three-Dimensional Photofabrication with Femtosecond Lasers for Applications in Photonics and Biomedicine. Applied Surface Science. 2007;253(15):6599-602.

Barcikowski S, Hahn A, Kabashin AV, Chichkov BN. Properties of Nanoparticles Generated During Femtosecond Laser Machining in Air and Water. Applied Physics A. 2007;87(1):47-55.

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