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Examination of Hydrogels and Mesenchymal Stem Cell Sources for Bioprinting of Artificial Osteogenic Tissues

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Abstract

Introduction

Mesenchymal stem cells (MSCs) represent a very important cell source in the field of regenerative medicine and for bone and cartilage tissue engineering applications. Three-dimensional (3D) bioprinting has the potential to improve the classical tissue engineering concept as this technique allows the printing of cells with high spatial control of cell allocation within a 3D construct. In this study, we systematically compared different hydrogel blends for 3D bioprinting of MSCs by testing their cytocompatibility, ability to support osteogenic differentiation and their mechanical properties. In addition, we compared four different MSC populations isolated from different human tissues for their osteogenic differentiation capacity in combination with different hydrogels. The aim of this study was to identify the best MSC source and the most suitable hydrogel blend for extrusion-based bioprinting of 3D large-scaled osteogenic constructs.

Materials and Methods

MSCs were isolated from different tissues (umbilical cord, adipose tissue, bone marrow). MSCs were seed onto or into different hydrogels and analyzed for cell viability, proliferation and osteogenic differentiation. In addition, viscoelastic properties of the hydrogels were determined. MSC-containing cubes with the size of 1 cm3 were printed by means of 3D extrusion-based bioprinting and analyzed by (immuno)histology for cell survival and production of a calcified extracellular matrix.

Results

Adipose tissue derived MSCs (ASCs) showed the highest osteogenic differentiation potential. A complex hydrogel blend consisting of fibrin, gelatin, hyaluronic acid, glycerol (F/G/H/Gl), tuned with hydroxyapatite, showed the best viscoelastic properties in combination with an excellent biocompatibility towards ASCs. This cell/hydrogel combination was used to bioprint 3D cubes. The cubes showed good mechanical stability and the printed ASCs were viable and able to calcify the hydrogel after bioprinting.

Conclusions

The combination of the HA-tuned F/G/H/Gl hydrogel blend along with ASCs can be considered as a very promising bioink for 3D bioprinting of artificial bone tissue equivalents for prospective applications in tissue engineering and regenerative medicine.

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Acknowledgments

This work was supported by funding through the Deutsche Forschungsgemeinschaft (FI 790/10-1 and KO 3910/1-1) and the Bundesministerium für Bildung und Forschung (03VNE1034C and 03VNE1034B). The authors would like to thank Brunhilde Baumer for excellent technical assistance, Matthias Schieker for providing immortalized mesenchymal stem cells and Anja Eisenhardt for critical reading of the manuscript.

Conflict of interest

Maximilian Wehrle, Fritz Koch, Stefan Zimmermann, Peter Koltay, Roland Zengerle, G. Björn Stark, Sandra Strassburg and Günter Finkenzeller declare that they have no conflicts of interest.

Ethical Approval

No human studies and no animal experiments were carried out by the authors of this article.

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Authors and Affiliations

  1. Department of Plastic and Hand Surgery, Medical Center University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetterstrasse 55, 79106, Freiburg, Germany

    Maximilian Wehrle, G. Björn Stark, Sandra Strassburg & Günter Finkenzeller

  2. Laboratory for MEMS Applications, IMTEK‐Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, 79110, Freiburg, Germany

    Fritz Koch, Stefan Zimmermann, Peter Koltay & Roland Zengerle

  3. Hahn-Schickard, Georges-Koehler-Allee 103, 79110, Freiburg, Germany

    Stefan Zimmermann, Peter Koltay & Roland Zengerle

  4. Freiburg Center for Interactive Materials and Bioinspired Technologies, University of Freiburg, Georges-Koehler-Allee 105, 79110, Freiburg, Germany

    Peter Koltay

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  1. Maximilian Wehrle
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Correspondence to Günter Finkenzeller.

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12195_2019_588_MOESM1_ESM.tif

SUPPLEMENT FIGURE S1. Flow cytometric analysis of iMSCs, ASCs, ucMSCs and bmMSCs. All MSC types were positive for CD105, CD90 and CD73 and negative for CD45, CD14 and HLA-DR. Supplementary material 1 (TIFF 370 kb).

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Wehrle, M., Koch, F., Zimmermann, S. et al. Examination of Hydrogels and Mesenchymal Stem Cell Sources for Bioprinting of Artificial Osteogenic Tissues. Cel. Mol. Bioeng. 12, 583–597 (2019). https://doi.org/10.1007/s12195-019-00588-x

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