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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 PPAR gamma 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 promotemore » BMP9-induced osteogenic differentiation of MSCs and hold promise for the development of bone tissue engineering platforms.« less

Authors:
 [1];  [2];  [3];  [2];  [4];  [4];  [4];  [5];  [6];  [7];  [4];  [4];  [4];  [6];  [4];  [4];  [4];  [4];  [7];  [4] more »;  [4];  [4];  [8];  [9];  [10];  [10];  [2];  [4]; ORCiD logo [2]; ORCiD logo [11] « less
  1. 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
  2. Institute for Molecular Engineering, The University of Chicago, Chicago, Illinois 60637, United States; Argonne National Laboratory, Argonne, Illinois 60439, United States
  3. Institute for Molecular Engineering, The University of Chicago, Chicago, Illinois 60637, United States
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
  10. Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center, Chicago, Illinois 60637, United States
  11. Department of Orthopaedic Surgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
National Institute of Standards and Technology (NIST) - Center for Hierarchical Materials Design (CHiMaD); National Natural Science Foundation of China (NNSFC); National Institutes of Health (NIH); USDOD; National Science Foundation (NSF)
OSTI Identifier:
1393461
DOE Contract Number:
AC02-06CH11357
Resource Type:
Journal Article
Resource Relation:
Journal Name: ACS Applied Materials and Interfaces; Journal Volume: 9; Journal Issue: 19
Country of Publication:
United States
Language:
English

Citation Formats

Zou, Yulong, Qazvini, Nader Taheri, Zane, Kylie, Sadati, Monirosadat, Wei, Qiang, Liao, Junyi, Fan, Jiaming, Song, Dongzhe, Liu, Jianxiang, Ma, Chao, Qu, Xiangyang, Chen, Liqun, Yu, Xinyi, Zhang, Zhicai, Zhao, Chen, Zeng, Zongyue, Zhang, Ruyi, Yan, Shujuan, Wu, Tingting, Wu, Xingye, Shu, Yi, Li, Yasha, Zhang, Wenwen, Reid, Russell R., Lee, Michael J., Wolf, Jennifer Moritis, Tirrell, Matthew, He, Tong-Chuan, de Pablo, Juan J., and Deng, Zhong-Liang. Gelatin-Derived Graphene–Silicate Hybrid Materials Are Biocompatible and Synergistically Promote BMP9-Induced Osteogenic Differentiation of Mesenchymal Stem Cells. United States: N. p., 2017. Web. doi:10.1021/acsami.7b00272.
Zou, Yulong, Qazvini, Nader Taheri, Zane, Kylie, Sadati, Monirosadat, Wei, Qiang, Liao, Junyi, Fan, Jiaming, Song, Dongzhe, Liu, Jianxiang, Ma, Chao, Qu, Xiangyang, Chen, Liqun, Yu, Xinyi, Zhang, Zhicai, Zhao, Chen, Zeng, Zongyue, Zhang, Ruyi, Yan, Shujuan, Wu, Tingting, Wu, Xingye, Shu, Yi, Li, Yasha, Zhang, Wenwen, Reid, Russell R., Lee, Michael J., Wolf, Jennifer Moritis, Tirrell, Matthew, He, Tong-Chuan, de Pablo, Juan J., & Deng, Zhong-Liang. Gelatin-Derived Graphene–Silicate Hybrid Materials Are Biocompatible and Synergistically Promote BMP9-Induced Osteogenic Differentiation of Mesenchymal Stem Cells. United States. doi:10.1021/acsami.7b00272.
Zou, Yulong, Qazvini, Nader Taheri, Zane, Kylie, Sadati, Monirosadat, Wei, Qiang, Liao, Junyi, Fan, Jiaming, Song, Dongzhe, Liu, Jianxiang, Ma, Chao, Qu, Xiangyang, Chen, Liqun, Yu, Xinyi, Zhang, Zhicai, Zhao, Chen, Zeng, Zongyue, Zhang, Ruyi, Yan, Shujuan, Wu, Tingting, Wu, Xingye, Shu, Yi, Li, Yasha, Zhang, Wenwen, Reid, Russell R., Lee, Michael J., Wolf, Jennifer Moritis, Tirrell, Matthew, He, Tong-Chuan, de Pablo, Juan J., and Deng, Zhong-Liang. Thu . "Gelatin-Derived Graphene–Silicate Hybrid Materials Are Biocompatible and Synergistically Promote BMP9-Induced Osteogenic Differentiation of Mesenchymal Stem Cells". United States. doi:10.1021/acsami.7b00272.
@article{osti_1393461,
title = {Gelatin-Derived Graphene–Silicate Hybrid Materials Are Biocompatible and Synergistically Promote BMP9-Induced Osteogenic Differentiation of Mesenchymal Stem Cells},
author = {Zou, Yulong and Qazvini, Nader Taheri and Zane, Kylie and Sadati, Monirosadat and Wei, Qiang and Liao, Junyi and Fan, Jiaming and Song, Dongzhe and Liu, Jianxiang and Ma, Chao and Qu, Xiangyang and Chen, Liqun and Yu, Xinyi and Zhang, Zhicai and Zhao, Chen and Zeng, Zongyue and Zhang, Ruyi and Yan, Shujuan and Wu, Tingting and Wu, Xingye and Shu, Yi and Li, Yasha and Zhang, Wenwen and Reid, Russell R. and Lee, Michael J. and Wolf, Jennifer Moritis and Tirrell, Matthew and He, Tong-Chuan and de Pablo, Juan J. and Deng, Zhong-Liang},
abstractNote = {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 PPAR gamma 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.},
doi = {10.1021/acsami.7b00272},
journal = {ACS Applied Materials and Interfaces},
number = 19,
volume = 9,
place = {United States},
year = {Thu May 04 00:00:00 EDT 2017},
month = {Thu May 04 00:00:00 EDT 2017}
}
  • Periodontitis is a chronic inflammatory disease induced by bacterial pathogens, which not only affect connective tissue attachments but also cause alveolar bone loss. In this study, we investigated the anti-inflammatory effects of Human amnion-derived mesenchymal stem cells (HAMSCs) on human bone marrow mesenchymal stem cells (HBMSCs) under lipopolysaccharide (LPS)-induced inflammatory conditions. Proliferation levels were measured by flow cytometry and immunofluorescence staining of 5-ethynyl-2′-deoxyuridine (EdU). Osteoblastic differentiation and mineralization were investigated using chromogenic alkaline phosphatase activity (ALP) activity substrate assays, Alizarin red S staining, and RT-PCR analysis of HBMSCs osteogenic marker expression. Oxidative stress induced by LPS was investigated by assayingmore » reactive oxygen species (ROS) level and superoxide dismutase (SOD) activity. Here, we demonstrated that HAMSCs increased the proliferation, osteoblastic differentiation, and SOD activity of LPS-induced HBMSCs, and down-regulated the ROS level. Moreover, our results suggested that the activation of p38 MAPK signal transduction pathway is essential for reversing the LPS-induced bone-destructive processes. SB203580, a selective inhibitor of p38 MAPK signaling, significantly suppressed the anti-inflammatory effects in HAMSCs. In conclusion, HAMSCs show a strong potential in treating inflammation-induced bone loss by influencing p38 MAPK signaling. - Highlights: • LPS inhibites osteogenic differentiation in HBMSCs via suppression of p38 MAPK signaling pathway. • HAMSCs promote LPS-induced HBMSCs osteogenic differentiation through p38 MAPK signaling pathway. • HAMSCs reverse LPS-induced oxidative stress in LPS-induced HBMSCs through p38 MAPK signaling pathway.« less
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