title: Osteogenic Differentiation of Three-Dimensional Bioprinted Constructs Consisting of Human Adipose-Derived Stem Cells In Vitro and In Vivo.
abstract: Here, we aimed to investigate osteogenic differentiation of human adipose-derived stem cells (hASCs) in three-dimensional (3D) bioprinted tissue constructs in vitro and in vivo. A 3D Bio-plotter dispensing system was used for building 3D constructs. Cell viability was determined using live/dead cell staining. After 7 and 14 days of culture, real-time quantitative polymerase chain reaction (PCR) was performed to analyze the expression of osteogenesis-related genes (RUNX2, OSX, and OCN). Western blotting for RUNX2 and immunofluorescent staining for OCN and RUNX2 were also performed. At 8 weeks after surgery, osteoids secreted by osteogenically differentiated cells were assessed by hematoxylin-eosin (H&E) staining, Masson trichrome staining, and OCN immunohistochemical staining. Results from live/dead cell staining showed that most of the cells remained alive, with a cell viability of 89%, on day 1 after printing. In vitro osteogenic induction of the 3D construct showed that the expression levels of RUNX2, OSX, and OCN were significantly increased on days 7 and 14 after printing in cells cultured in osteogenic medium (OM) compared with that in normal proliferation medium (PM). Fluorescence microscopy and western blotting showed that the expression of osteogenesis-related proteins was significantly higher in cells cultured in OM than in cells cultured in PM. In vivo studies demonstrated obvious bone matrix formation in the 3D bioprinted constructs. These results indicated that 3D bioprinted constructs consisting of hASCs had the ability to promote mineralized matrix formation and that hASCs could be used in 3D bioprinted constructs for the repair of large bone tissue defects.
authors.affiliation: Center of Digital Dentistry, Faculty of Prosthodontics, Peking University School and Hospital of Stomatology & National Engineering Laboratory for Digital and Material Technology of Stomatology & Research Center of Engineering and Technology for Digital Dentistry of Ministry of Health, Beijing 100081, China.
authors.forename: Xiao-Fei
authors.initials: XF
authors.lastname: Wang
authors.affiliation: Center of Digital Dentistry, Faculty of Prosthodontics, Peking University School and Hospital of Stomatology & National Engineering Laboratory for Digital and Material Technology of Stomatology & Research Center of Engineering and Technology for Digital Dentistry of Ministry of Health, Beijing 100081, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Song
authors.affiliation: Center of Digital Dentistry, Faculty of Prosthodontics, Peking University School and Hospital of Stomatology & National Engineering Laboratory for Digital and Material Technology of Stomatology & Research Center of Engineering and Technology for Digital Dentistry of Ministry of Health, Beijing 100081, China.
authors.forename: Yun-Song
authors.initials: YS
authors.lastname: Liu
authors.affiliation: Center of Digital Dentistry, Faculty of Prosthodontics, Peking University School and Hospital of Stomatology & National Engineering Laboratory for Digital and Material Technology of Stomatology & Research Center of Engineering and Technology for Digital Dentistry of Ministry of Health, Beijing 100081, China.
authors.forename: Yu-Chun
authors.initials: YC
authors.lastname: Sun
authors.affiliation: Center of Digital Dentistry, Faculty of Prosthodontics, Peking University School and Hospital of Stomatology & National Engineering Laboratory for Digital and Material Technology of Stomatology & Research Center of Engineering and Technology for Digital Dentistry of Ministry of Health, Beijing 100081, China.
authors.forename: Yu-Guang
authors.initials: YG
authors.lastname: Wang
authors.affiliation: Center of Digital Dentistry, Faculty of Prosthodontics, Peking University School and Hospital of Stomatology & National Engineering Laboratory for Digital and Material Technology of Stomatology & Research Center of Engineering and Technology for Digital Dentistry of Ministry of Health, Beijing 100081, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Center of Digital Dentistry, Faculty of Prosthodontics, Peking University School and Hospital of Stomatology & National Engineering Laboratory for Digital and Material Technology of Stomatology & Research Center of Engineering and Technology for Digital Dentistry of Ministry of Health, Beijing 100081, China.
authors.forename: Pei-Jun
authors.initials: PJ
authors.lastname: Lyu
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000464:Alginates / Q000494:pharmacology; D062028:Bioprinting / Q000379:methods*; D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002828:Choristoma / Q000473:pathology; D005455:Fluorescent Antibody Technique; D005780:Gelatin / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D020723:Glucuronic Acid / Q000494:pharmacology; D006603:Hexuronic Acids / Q000494:pharmacology; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000494:pharmacology; D010012:Osteogenesis / Q000187:drug effects; D016062:Porosity; D066330:Printing, Three-Dimensional; D060888:Real-Time Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D020723:Glucuronic Acid; D005780:Gelatin