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Investigation of osteoblast-like MC3T3-E1 cells on a novel recombinant collagen-like protein surface with triple helix structure

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Abstract

Fol-8Col is a novel recombinant collagen-like protein incorporated with foldon sequences derived from the native T4 phage fibritin. In this paper, we examined the potential of using Fol-8Col as scaffold for bone tissue engineering. Circular dichroism (CD) spectra indicate that the triple helix structure of Fol-8Col exists at temperatures ranging from 4 to 40 °C. Lactate dehydrogenase assay results and live/death cell staining of osteoblast-like MC3T3-E1 cells, cultivated on Fol-8Col for 24 h, showed evidence of cell cytocompatibility comparable to that of native type I collagen. Attachment and spreading of osteoblast-like MC3T3-E1 cells seeded on Fol-8Col were studied by immunofluorescence staining of cell nuclei, vinculin, and F-actin. Intensive focal adhesion patches and an elongated cortical actin cytoskeleton were observed after 24 hours’ cultivation. Proliferation assays of MC3T3-E1 cells cultivated on Fol-8Col for 2 weeks revealed no consistent differences in rate and pattern compared to growth on type I collagen. Alkaline phosphatase activity assay and osteogenic gene expression, detected by RT-PCR, evaluated the osteogenic differentiation of MC3T3-E1 cells on Fol-8Col. This study shows that Fol-8Col, with a triple helix structure, has good potential for application in bone regeneration as a replacement for native collagen, thereby reducing the risk of contamination.

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Acknowledgements

This work was supported by a Grant-in-Aid for Global COE Program by the Ministry of Education, Culture, Sports, Science and Technology.

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Authors and Affiliations

  1. Department of Functional Polymer Science, Faculty of Textile Science and Technology, Shinshu University, Ueda, Nagano, 386-3485, Japan

    Yuan Li, Xu Xu Bao, Naoki Matsuda, Akira Teramoto & Koji Abe

  2. The Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, College of Materials and Textile, Zhejiang Sci-Tech University, Hangzhou, 310018, People’s Republic of China

    Ju Ming Yao

  3. Advanced Fibrous Materials, University of British Columbia, Vancouver, Canada

    Frank K. Ko

Authors
  1. Yuan Li
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  2. Xu Xu Bao
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  3. Naoki Matsuda
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  4. Ju Ming Yao
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  5. Akira Teramoto
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  6. Koji Abe
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  7. Frank K. Ko
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Correspondence to Yuan Li.

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Li, Y., Bao, X.X., Matsuda, N. et al. Investigation of osteoblast-like MC3T3-E1 cells on a novel recombinant collagen-like protein surface with triple helix structure. J Mater Sci 46, 1396–1404 (2011). https://doi.org/10.1007/s10853-010-4933-2

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  • DOI: https://doi.org/10.1007/s10853-010-4933-2

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