title: Differentiation of adipose stem cells seeded towards annulus fibrosus cells on a designed poly(trimethylene carbonate) scaffold prepared by stereolithography.
abstract: Cell-based therapies could potentially restore the biomechanical function and enhance the self-repair capacity of annulus fibrosus (AF) tissue. However, choosing a suitable cell source and scaffold design are still key challenges. In this study, we assessed the in vitro ability of human adipose stem cells (hASCs), an easily available cell source to produce AF-like matrix in novel AF-mimetic designed scaffolds based on poly(trimethylene carbonate) and built by stereolithography. To facilitate efficient differentiation of hASCs towards AF tissue, we tested different culture medium compositions and cell seeding techniques. This is the first study to report that medium supplementation with transforming growth factor (TGF)-β3 is essential to support AF differentiation of hASCs while TGF-β1 has negligible effect after 21 days of culture. Fibrin gel seeding resulted in superior cell distribution, proliferation and AF-like matrix production of hASCs compared to direct and micromass seeding under TGF-β3 stimulation. Not only the production of sulphated glycosaminoglycans (sGAG) and collagen was significantly upregulated, but the formed collagen was also oriented and aligned into bundles within the designed pore channels. The differentiated hASCs seeded with fibrin gel were also found to have a comparable sGAG:collagen ratio and gene expression profile as native AF cells demonstrating the high potential of this strategy in AF repair. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: MIRA Institute for Biomedical Technology and Technical Medicine, Department of Biomaterials Science and Technology, University of Twente, Enschede, the Netherlands.|Collaborative Research Partner Annulus Fibrosus Rupture Program of AO Foundation, Davos, Switzerland.
authors.forename: Sébastien B G
authors.initials: SBG
authors.lastname: Blanquer
authors.affiliation: MIRA Institute for Biomedical Technology and Technical Medicine, Department of Biomaterials Science and Technology, University of Twente, Enschede, the Netherlands.|Department of Oral and Maxillofacial Sciences, Clinicum, University of Helsinki, Helsinki, Finland.
authors.forename: Arjen W H
authors.initials: AWH
authors.lastname: Gebraad
authors.affiliation: Institute of Biosciences and Medical Technology (BioMediTech), University of Tampere, Tampere, Finland.
authors.forename: Susanna
authors.initials: S
authors.lastname: Miettinen
authors.affiliation: MIRA Institute for Biomedical Technology and Technical Medicine, Department of Biomaterials Science and Technology, University of Twente, Enschede, the Netherlands.|Collaborative Research Partner Annulus Fibrosus Rupture Program of AO Foundation, Davos, Switzerland.
authors.forename: André A
authors.initials: AA
authors.lastname: Poot
authors.affiliation: MIRA Institute for Biomedical Technology and Technical Medicine, Department of Biomaterials Science and Technology, University of Twente, Enschede, the Netherlands.|Collaborative Research Partner Annulus Fibrosus Rupture Program of AO Foundation, Davos, Switzerland.|University of Groningen, University Medical Centre Groningen, W.J. Kolff Institute, Department of Biomedical Engineering, Groningen, the Netherlands.
authors.forename: Dirk W
authors.initials: DW
authors.lastname: Grijpma
authors.affiliation: MIRA Institute for Biomedical Technology and Technical Medicine, Department of Biomaterials Science and Technology, University of Twente, Enschede, the Netherlands.|Department of Oral and Maxillofacial Sciences, Clinicum, University of Helsinki, Helsinki, Finland.
authors.forename: Suvi P
authors.initials: SP
authors.lastname: Haimi
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000070616:Annulus Fibrosus / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects; D003094:Collagen / Q000737:chemistry; D004247:DNA / Q000378:metabolism; D004146:Dioxanes / Q000494:pharmacology*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008751:Methylene Blue / Q000737:chemistry; D008875:Middle Aged; D011108:Polymers / Q000494:pharmacology*; D013194:Staining and Labeling; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D000076227:Stereolithography; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D004146:Dioxanes; D011108:Polymers; C059299:polytrimethylene carbonate; D003094:Collagen; D004247:DNA; D008751:Methylene Blue
keywords: AF-mimetic designed scaffold; annulus fibrosus; human adipose stem cells (hASCs); oriented collagen fibres; poly(trimethylene carbonate) PTMC; stereolithography; tissue engineering