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Thiol-gelatin-norbornene bioink for laser‐based high‐definition bioprinting

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Abstract
Two-photon polymerization (2PP) is a lithography-based 3D printing method allowing the fabrication of 3D structures with sub-micrometer resolution. This work focuses on the characterization of gelatin-norbornene (Gel-NB) bioinks which enables the embedding of cells via 2PP. The high reactivity of the thiol-ene system allows 2PP processing of cell-containing materials at remarkably high scanning speeds (1000 mm s(-1)) placing this technology in the domain of bioprinting. Atomic force microscopy results demonstrate that the indentation moduli of the produced hydrogel constructs can be adjusted in the 0.2-0.7 kPa range by controlling the 2PP processing parameters. Using this approach gradient 3D constructs are produced and the morphology of the embedded cells is observed in the course of 3 weeks. Furthermore, it is possible to tune the enzymatic degradation of the crosslinked bioink by varying the applied laser power. The 3D printed Gel-NB hydrogel constructs show exceptional biocompatibility, supported cell adhesion, and migration. Furthermore, cells maintain their proliferation capacity demonstrated by Ki-67 immunostaining. Moreover, the results demonstrate that direct embedding of cells provides uniform distribution and high cell loading independently of the pore size of the scaffold. The investigated photosensitive bioink enables high-definition bioprinting of well-defined constructs for long-term cell culture studies.
Keywords
gelatin, high-resolution bioprinting, hydrogels, multiphoton lithography, thiol-ene chemistry, CELL-CULTURE, HYDROGELS, PROTEIN, CALIBRATION, INITIATOR, MATRICES

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MLA
Dobos, Agnes, et al. “Thiol-Gelatin-Norbornene Bioink for Laser‐based High‐definition Bioprinting.” ADVANCED HEALTHCARE MATERIALS, vol. 9, no. 15, 2020, doi:10.1002/adhm.201900752.
APA
Dobos, A., Van Hoorick, J., Steiger, W., Gruber, P., Markovic, M., Andriotis, O. G., … Ovsianikov, A. (2020). Thiol-gelatin-norbornene bioink for laser‐based high‐definition bioprinting. ADVANCED HEALTHCARE MATERIALS, 9(15). https://doi.org/10.1002/adhm.201900752
Chicago author-date
Dobos, Agnes, Jasper Van Hoorick, Wolfgang Steiger, Peter Gruber, Marica Markovic, Orestis G Andriotis, Andreas Rohatschek, et al. 2020. “Thiol-Gelatin-Norbornene Bioink for Laser‐based High‐definition Bioprinting.” ADVANCED HEALTHCARE MATERIALS 9 (15). https://doi.org/10.1002/adhm.201900752.
Chicago author-date (all authors)
Dobos, Agnes, Jasper Van Hoorick, Wolfgang Steiger, Peter Gruber, Marica Markovic, Orestis G Andriotis, Andreas Rohatschek, Peter Dubruel, Philipp J Thurner, Sandra Van Vlierberghe, Stefan Baudis, and Aleksandr Ovsianikov. 2020. “Thiol-Gelatin-Norbornene Bioink for Laser‐based High‐definition Bioprinting.” ADVANCED HEALTHCARE MATERIALS 9 (15). doi:10.1002/adhm.201900752.
Vancouver
1.
Dobos A, Van Hoorick J, Steiger W, Gruber P, Markovic M, Andriotis OG, et al. Thiol-gelatin-norbornene bioink for laser‐based high‐definition bioprinting. ADVANCED HEALTHCARE MATERIALS. 2020;9(15).
IEEE
[1]
A. Dobos et al., “Thiol-gelatin-norbornene bioink for laser‐based high‐definition bioprinting,” ADVANCED HEALTHCARE MATERIALS, vol. 9, no. 15, 2020.
@article{8623949,
  abstract     = {{Two-photon polymerization (2PP) is a lithography-based 3D printing method allowing the fabrication of 3D structures with sub-micrometer resolution. This work focuses on the characterization of gelatin-norbornene (Gel-NB) bioinks which enables the embedding of cells via 2PP. The high reactivity of the thiol-ene system allows 2PP processing of cell-containing materials at remarkably high scanning speeds (1000 mm s(-1)) placing this technology in the domain of bioprinting. Atomic force microscopy results demonstrate that the indentation moduli of the produced hydrogel constructs can be adjusted in the 0.2-0.7 kPa range by controlling the 2PP processing parameters. Using this approach gradient 3D constructs are produced and the morphology of the embedded cells is observed in the course of 3 weeks. Furthermore, it is possible to tune the enzymatic degradation of the crosslinked bioink by varying the applied laser power. The 3D printed Gel-NB hydrogel constructs show exceptional biocompatibility, supported cell adhesion, and migration. Furthermore, cells maintain their proliferation capacity demonstrated by Ki-67 immunostaining. Moreover, the results demonstrate that direct embedding of cells provides uniform distribution and high cell loading independently of the pore size of the scaffold. The investigated photosensitive bioink enables high-definition bioprinting of well-defined constructs for long-term cell culture studies.}},
  articleno    = {{1900752}},
  author       = {{Dobos, Agnes and Van Hoorick, Jasper and Steiger, Wolfgang and Gruber, Peter and Markovic, Marica and Andriotis, Orestis G and Rohatschek, Andreas and Dubruel, Peter and Thurner, Philipp J and Van Vlierberghe, Sandra and Baudis, Stefan and Ovsianikov, Aleksandr}},
  issn         = {{2192-2640}},
  journal      = {{ADVANCED HEALTHCARE MATERIALS}},
  keywords     = {{gelatin,high-resolution bioprinting,hydrogels,multiphoton lithography,thiol-ene chemistry,CELL-CULTURE,HYDROGELS,PROTEIN,CALIBRATION,INITIATOR,MATRICES}},
  language     = {{eng}},
  number       = {{15}},
  pages        = {{9}},
  title        = {{Thiol-gelatin-norbornene bioink for laser‐based high‐definition bioprinting}},
  url          = {{http://dx.doi.org/10.1002/adhm.201900752}},
  volume       = {{9}},
  year         = {{2020}},
}

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