title: Gelatin-Derived Graphene-Silicate Hybrid Materials Are Biocompatible and Synergistically Promote BMP9-Induced Osteogenic Differentiation of Mesenchymal Stem Cells.
abstract: Graphene-based materials are used in many fields but have found only limited applications in biomedicine, including bone tissue engineering. Here, we demonstrate that novel hybrid materials consisting of gelatin-derived graphene and silicate nanosheets of Laponite (GL) are biocompatible and promote osteogenic differentiation of mesenchymal stem cells (MSCs). Homogeneous cell attachment, long-term proliferation, and osteogenic differentiation of MSCs on a GL-scaffold were confirmed using optical microscopy and scanning electron microscopy. GL-powders made by pulverizing the GL-scaffold were shown to promote bone morphogenetic protein (BMP9)-induced osteogenic differentiation. GL-powders increased the alkaline phosphatase (ALP) activity in immortalized mouse embryonic fibroblasts but decreased the ALP activity in more-differentiated immortalized mouse adipose-derived cells. Note, however, that GL-powders promoted BMP9-induced calcium mineral deposits in both MSC lines, as assessed using qualitative and quantitative alizarin red assays. Furthermore, the expression of chondro-osteogenic regulator markers such as Runx2, Sox9, osteopontin, and osteocalcin was upregulated by the GL-powder, independent of BMP9 stimulation; although the powder synergistically upregulated the BMP9-induced Osterix expression, the adipogenic marker PPARγ was unaffected. Furthermore, in vivo stem cell implantation experiments demonstrated that GL-powder could significantly enhance the BMP9-induced ectopic bone formation from MSCs. Collectively, our results strongly suggest that the GL hybrid materials promote BMP9-induced osteogenic differentiation of MSCs and hold promise for the development of bone tissue engineering platforms.
authors.affiliation: Department of Orthopaedic Surgery, The Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010, China.|Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.
authors.forename: Yulong
authors.initials: Y
authors.lastname: Zou
authors.affiliation: Institute for Molecular Engineering, The University of Chicago , Chicago, Illinois 60637, United States.|Argonne National Laboratory , Argonne, Illinois 60439, United States.
authors.forename: Nader Taheri
authors.initials: NT
authors.lastname: Qazvini
authors.affiliation: Institute for Molecular Engineering, The University of Chicago , Chicago, Illinois 60637, United States.
authors.forename: Kylie
authors.initials: K
authors.lastname: Zane
authors.affiliation: Institute for Molecular Engineering, The University of Chicago , Chicago, Illinois 60637, United States.|Argonne National Laboratory , Argonne, Illinois 60439, United States.
authors.forename: Monirosadat
authors.initials: M
authors.lastname: Sadati
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Wei
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Junyi
authors.initials: J
authors.lastname: Liao
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Jiaming
authors.initials: J
authors.lastname: Fan
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Department of Conservative Dentistry and Endodontics, West China School of Stomatology, Sichuan University , Chengdu 610041, China.
authors.forename: Dongzhe
authors.initials: D
authors.lastname: Song
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Department of Orthopaedic Surgery, Union Hospital of Tongji Medical College, Huazhong University of Science & Technology , Wuhan 430022, China.
authors.forename: Jianxiang
authors.initials: J
authors.lastname: Liu
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Departments of Neurosurgery and Otolaryngology-Head & Neck Surgery, The Affiliated Zhongnan Hospital of Wuhan University , Wuhan 430071, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Ma
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Xiangyang
authors.initials: X
authors.lastname: Qu
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Liqun
authors.initials: L
authors.lastname: Chen
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Xinyi
authors.initials: X
authors.lastname: Yu
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Department of Orthopaedic Surgery, Union Hospital of Tongji Medical College, Huazhong University of Science & Technology , Wuhan 430022, China.
authors.forename: Zhicai
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Zhao
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Zongyue
authors.initials: Z
authors.lastname: Zeng
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Ruyi
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Shujuan
authors.initials: S
authors.lastname: Yan
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Departments of Neurosurgery and Otolaryngology-Head & Neck Surgery, The Affiliated Zhongnan Hospital of Wuhan University , Wuhan 430071, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Wu
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Xingye
authors.initials: X
authors.lastname: Wu
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Shu
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Yasha
authors.initials: Y
authors.lastname: Li
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Department of Laboratory Medicine and Clinical Diagnostics, the Affiliated Yantai Hospital, Binzhou Medical University , Yantai 264100, China.
authors.forename: Wenwen
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Department of Surgery, Section of Plastic Surgery, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.
authors.forename: Russell R
authors.initials: RR
authors.lastname: Reid
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Lee
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.
authors.forename: Jennifer Moritis
authors.initials: JM
authors.lastname: Wolf
authors.affiliation: Institute for Molecular Engineering, The University of Chicago , Chicago, Illinois 60637, United States.|Argonne National Laboratory , Argonne, Illinois 60439, United States.
authors.forename: Matthew
authors.initials: M
authors.lastname: Tirrell
authors.affiliation: Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center , Chicago, Illinois 60637, United States.|Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Tong-Chuan
authors.initials: TC
authors.lastname: He
authors.affiliation: Institute for Molecular Engineering, The University of Chicago , Chicago, Illinois 60637, United States.|Argonne National Laboratory , Argonne, Illinois 60439, United States.
authors.forename: Juan J
authors.identifier: http://orcid.org/0000-0002-3526-516X
authors.initials: JJ
authors.lastname: de Pablo
authors.affiliation: Department of Orthopaedic Surgery, The Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010, China.
authors.forename: Zhong-Liang
authors.identifier: http://orcid.org/0000-0003-4831-1775
authors.initials: ZL
authors.lastname: Deng
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D005780:Gelatin; D006108:Graphite; D055427:Growth Differentiation Factor 2; D059630:Mesenchymal Stem Cells; D051379:Mice; D010012:Osteogenesis; D017640:Silicates
chemical_list: C526108:Gdf2 protein, mouse; D055427:Growth Differentiation Factor 2; D017640:Silicates; D006108:Graphite; D005780:Gelatin
keywords: Laponite; bone morphogenetic protein; bone tissue engineering; graphene; mesenchymal stem cells