Two-photon polymerization is a technique that involves simultaneous absorption of two photons from a femtosecond laser for selective polymerization of a photosensitive material. In this study, two-photon polymerization was used for layer-by-layer fabrication of 3-D scaffolds composed of an inorganic–organic zirconium oxide hybrid material. Four types of scaffold microarchitectures were created, which exhibit layers of parallel line features at various orientations as well as pores between the line features. Long-term cell culture studies involving human bone marrow stromal cells were conducted using these 3-D scaffolds. Cellular adhesion and proliferation were demonstrated on all of the scaffold types; tissuelike structure was shown to span the pores. This study indicates that two-photon polymerization may be used to create microstructured scaffolds out of an inorganic–organic zirconium oxide hybrid material for use in 3-D tissue culture systems.
The authors would like to acknowledge the FDA White Oak Nanotechnology Core Facility for instrument use and scientific and technical assistance. In addition, the authors would like to acknowledge financial support from U.S. National Science Foundation Award #1041375. The authors would also like to thank the Laser Zentrum Hannover (LZH) in Hannover, Germany for use of their two-photon polymerization system for fabrication procedures. A.K. and B.N.C. acknowledge support by the German Research Foundation Cluster of Excellence Rebirth “From Regenerative Biology to Reconstructive Therapy” and the Lower Saxony project “Biofabrication for NIFE.” The mention of commercial products, their sources, or their use in connection with material reported herein is not to be construed as either an actual or implied endorsement of such products by the Department of Health and Human Services.
A. Zirconium oxide hybrid material synthesis and film preparation
B. Two-photon polymerization and postprocessing of scaffolds
C. Physicochemical characterization: SEM, AFM, and EDS
D. Cell culture and cell seeding of scaffolds
E. Preparation and SEM imaging of cells on scaffolds
III. RESULTS AND DISCUSSION
A. Two-photon polymerization of zirconium oxide hybrid material scaffolds
B. Cellular response to 2PP-fabricated zirconium oxide hybrid material microstructured scaffolds
IV. SUMMARY AND CONCLUSIONS
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