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Assessment of cell sheets derived from human periodontal ligament cells: a pre-clinical study

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Abstract

Periodontal-ligament-derived cells (PDL cells) have stem-cell-like properties and, when implanted into periodontal defects in vivo, can induce periodontal regeneration including the formation of new bone, cementum, and periodontal ligament. We have previously demonstrated that PDL cell sheets, harvested from temperature-responsive cell culture dishes, have a great potential for periodontal regeneration. The purpose of this study has been to validate the safety and efficacy of human PDL (hPDL) cell sheets for use in clinical trials. hPDL tissues from three donors were enzymatically digested, and the obtained cells were cultured with media containing autologous serum in a cell-processing center (CPC). The safety and efficacy of hPDL cell sheets were evaluated both in vitro and in vivo. In vitro studies showed that the hPDL cell sheets had high alkaline phosphatase activity and periostin expression (known PDL markers) and no contamination with microorganisms. In vivo studies revealed that hPDL cell sheets, implanted with dentin blocks, induced the formation of cementum and PDL-like tissue in immunodeficient mice. The hPDL cells presented no evidence of malignant transformation. Thus, hPDL cell sheets created in CPCs are safe products and possess the potential to regenerate periodontal tissues.

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Acknowledgements

The authors thank Dr. Toshiyuki Yoshida, Ms. Hazuki Samejima, Mr. Hiroaki Sugiyama, Ms. Tomoka Izawa, and Mr. Shigeki Mukobata for their assistance.

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Authors and Affiliations

  1. Institute of Advanced Biomedical Engineering and Science, Tokyo Women’s Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo, 162-8666, Japan

    Kaoru Washio, Takanori Iwata, Masayuki Yamato, Teruo Okano & Isao Ishikawa

  2. Department of Oral and Maxillofacial Surgery, Tokyo Women’s Medical University, Graduate School of Medicine, 8-1 Kawada-cho, Shinjuku-ku, Tokyo, 162-8666, Japan

    Takanori Iwata, Tomohiro Ando & Isao Ishikawa

  3. CellSeed, 33-8 Wakamatsu-cho, Shinjuku-ku, Tokyo, 162-0056, Japan

    Manabu Mizutani

Authors
  1. Kaoru Washio
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  2. Takanori Iwata
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  3. Manabu Mizutani
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  4. Tomohiro Ando
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  5. Masayuki Yamato
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  6. Teruo Okano
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  7. Isao Ishikawa
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Corresponding authors

Correspondence to Kaoru Washio or Teruo Okano.

Additional information

This study was supported by the Formation of Innovation Center for Fusion of Advanced Technologies in the Special Coordination Funds for Promoting Science and Technology from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT), Japan.

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Washio, K., Iwata, T., Mizutani, M. et al. Assessment of cell sheets derived from human periodontal ligament cells: a pre-clinical study. Cell Tissue Res 341, 397–404 (2010). https://doi.org/10.1007/s00441-010-1009-1

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  • DOI: https://doi.org/10.1007/s00441-010-1009-1

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