Kaisa Vuornos1, Miina Ojansivu2, Janne T Koivisto3, Heikki Häkkänen4, Birhanu Belay5, Toni Montonen6, Heini Huhtala7, Minna Kääriäinen8, Leena Hupa9, Minna Kellomäki10, Jari Hyttinen11, Janne A Ihalainen12, Susanna Miettinen13. 1. Adult Stem Cell Group, BioMediTech, Faculty of Medicine and Health Technology, Tampere University, P.O. BOX 100, FI-33014 Tampere, Finland; Research, Development and Innovation Centre, Tampere University Hospital, P.O. BOX 2000, FI-33521, Tampere, Finland. Electronic address: kaisa.vuornos@tuni.fi. 2. Adult Stem Cell Group, BioMediTech, Faculty of Medicine and Health Technology, Tampere University, P.O. BOX 100, FI-33014 Tampere, Finland; Research, Development and Innovation Centre, Tampere University Hospital, P.O. BOX 2000, FI-33521, Tampere, Finland. Electronic address: miina.ojansivu@ki.se. 3. Biomaterials and Tissue Engineering Group, BioMediTech, Faculty of Medicine and Health Technology, Tampere University, P.O. BOX 527, FI-33101 Tampere, Finland; Heart Group, BioMediTech, Faculty of Medicine and Health Technology, Tampere University, P.O. BOX 100, FI-33014 Tampere, Finland. Electronic address: janne.t.koivisto@tuni.fi. 4. Nanoscience Center, University of Jyväskylä, P.O. BOX 35, FI-40014 Jyväskylä, Finland. Electronic address: heikki.hakkanen@jyu.fi. 5. Computational Biophysics and Imaging Group, BioMediTech, Faculty of Medicine and Health Technology, Tampere University, P.O. BOX 527, FI-33101 Tampere, Finland. Electronic address: birhanu.belay@tuni.fi. 6. Computational Biophysics and Imaging Group, BioMediTech, Faculty of Medicine and Health Technology, Tampere University, P.O. BOX 527, FI-33101 Tampere, Finland. Electronic address: toni.montonen@tuni.fi. 7. Faculty of Social Sciences, Tampere University, P.O. BOX 100, FI-33014 Tampere, Finland. Electronic address: heini.huhtala@tuni.fi. 8. Department of Plastic and Reconstructive Surgery, Tampere University Hospital, P.O. BOX 2000, FI-33521 Tampere, Finland. Electronic address: minna.kaariainen@pshp.fi. 9. Johan Gadolin Process Chemistry Centre, Åbo Akademi University, Biskopsgatan 8, FI-20500 Åbo, Finland. Electronic address: leena.hupa@abo.fi. 10. Biomaterials and Tissue Engineering Group, BioMediTech, Faculty of Medicine and Health Technology, Tampere University, P.O. BOX 527, FI-33101 Tampere, Finland. Electronic address: minna.kellomaki@tuni.fi. 11. Computational Biophysics and Imaging Group, BioMediTech, Faculty of Medicine and Health Technology, Tampere University, P.O. BOX 527, FI-33101 Tampere, Finland. Electronic address: jari.hyttinen@tuni.fi. 12. Nanoscience Center, University of Jyväskylä, P.O. BOX 35, FI-40014 Jyväskylä, Finland. Electronic address: janne.ihalainen@jyu.fi. 13. Adult Stem Cell Group, BioMediTech, Faculty of Medicine and Health Technology, Tampere University, P.O. BOX 100, FI-33014 Tampere, Finland; Research, Development and Innovation Centre, Tampere University Hospital, P.O. BOX 2000, FI-33521, Tampere, Finland. Electronic address: susanna.miettinen@tuni.fi.
Abstract
BACKGROUND: Due to unmet need for bone augmentation, our aim was to promote osteogenic differentiation of human adipose stem cells (hASCs) encapsulated in gellan gum (GG) or collagen type I (COL) hydrogels with bioactive glass (experimental glass 2-06 of composition [wt-%]: Na2O 12.1, K2O 14.0, CaO 19.8, P2O5 2.5, B2O3 1.6, SiO2 50.0) extract based osteogenic medium (BaG OM) for bone construct development. GG hydrogels were crosslinked with spermidine (GG-SPD) or BaG extract (GG-BaG). METHODS: Mechanical properties of cell-free GG-SPD, GG-BaG, and COL hydrogels were tested in osteogenic medium (OM) or BaG OM at 0, 14, and 21 d. Hydrogel embedded hASCs were cultured in OM or BaG OM for 3, 14, and 21 d, and analyzed for viability, cell number, osteogenic gene expression, osteocalcin production, and mineralization. Hydroxyapatite-stained GG-SPD samples were imaged with Optical Projection Tomography (OPT) and Selective Plane Illumination Microscopy (SPIM) in OM and BaG OM at 21 d. Furthermore, Raman spectroscopy was used to study the calcium phosphate (CaP) content of hASC-secreted ECM in GG-SPD, GG-BaG, and COL at 21 d in BaG OM. RESULTS: The results showed viable rounded cells in GG whereas hASCs were elongated in COL. Importantly, BaG OM induced significantly higher cell number and higher osteogenic gene expression in COL. In both hydrogels, BaG OM induced strong mineralization confirmed as CaP by Raman spectroscopy and significantly improved mechanical properties. GG-BaG hydrogels rescued hASC mineralization in OM. OPT and SPIM showed homogeneous 3D cell distribution with strong mineralization in BaG OM. Also, strong osteocalcin production was visible in COL. CONCLUSIONS: Overall, we showed efficacious osteogenesis of hASCs in 3D hydrogels with BaG OM with potential for bone-like grafts.
BACKGROUND: Due to unmet need for bone augmentation, our aim was to promote osteogenic differentiation of human adipose stem cells (hASCs) encapsulated in gellan gum (GG) or collagen type I (COL) hydrogels with bioactive glass (experimental glass 2-06 of composition [wt-%]: Na2O 12.1, K2O 14.0, CaO 19.8, P2O5 2.5, B2O3 1.6, SiO2 50.0) extract based osteogenic medium (BaG OM) for bone construct development. GG hydrogels were crosslinked with spermidine (GG-SPD) or BaG extract (GG-BaG). METHODS: Mechanical properties of cell-free GG-SPD, GG-BaG, and COL hydrogels were tested in osteogenic medium (OM) or BaG OM at 0, 14, and 21 d. Hydrogel embedded hASCs were cultured in OM or BaG OM for 3, 14, and 21 d, and analyzed for viability, cell number, osteogenic gene expression, osteocalcin production, and mineralization. Hydroxyapatite-stained GG-SPD samples were imaged with Optical Projection Tomography (OPT) and Selective Plane Illumination Microscopy (SPIM) in OM and BaG OM at 21 d. Furthermore, Raman spectroscopy was used to study the calcium phosphate (CaP) content of hASC-secreted ECM in GG-SPD, GG-BaG, and COL at 21 d in BaG OM. RESULTS: The results showed viable rounded cells in GG whereas hASCs were elongated in COL. Importantly, BaG OM induced significantly higher cell number and higher osteogenic gene expression in COL. In both hydrogels, BaG OM induced strong mineralization confirmed as CaP by Raman spectroscopy and significantly improved mechanical properties. GG-BaG hydrogels rescued hASC mineralization in OM. OPT and SPIM showed homogeneous 3D cell distribution with strong mineralization in BaG OM. Also, strong osteocalcin production was visible in COL. CONCLUSIONS: Overall, we showed efficacious osteogenesis of hASCs in 3D hydrogels with BaG OM with potential for bone-like grafts.
Authors: Birhanu Belay; Janne T Koivisto; Jenny Parraga; Olli Koskela; Toni Montonen; Minna Kellomäki; Edite Figueiras; Jari Hyttinen Journal: Sci Rep Date: 2021-03-22 Impact factor: 4.379
Authors: Martina Oriano; Laura Zorzetto; Giuseppe Guagliano; Federico Bertoglio; Sebastião van Uden; Livia Visai; Paola Petrini Journal: Front Bioeng Biotechnol Date: 2020-10-20