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

• Published in: Cell and tissue research 2010-09, Vol.341 (3), p.397-404
• Main Authors: Washio, Kaoru, Iwata, Takanori, Mizutani, Manabu, Ando, Tomohiro, Yamato, Masayuki, Okano, Teruo, Ishikawa, Isao
• Format: Article
• Language: English
• Subjects: Animals; Biomedical and Life Sciences; Biomedicine; Cell culture; Cell Culture Techniques - standards; Cell implantation; Cell sheet; Cells, Cultured; Cellular biology; Clinical trials; Dentin - cytology; Dentin - metabolism; Dentin - physiology; Feasibility Studies; Gene expression; Guided Tissue Regeneration - instrumentation; Guided Tissue Regeneration - methods; Guided Tissue Regeneration - standards; Human Genetics; Humans; Ligaments; Male; Mice; Mice, Inbred C57BL; Mice, Inbred ICR; Mice, SCID; Models, Biological; Molecular Medicine; Periodontal ligament; Periodontal Ligament - cytology; Periodontal Ligament - physiology; Periodontal Ligament - transplantation; Proteomics; Quality Control; Quality control test; regeneration; Regular Article; Tissue engineering; Tissue Scaffolds - standards
• ISSN: 0302-766X; 1432-0878
• DOI: 10.1007/s00441-010-1009-1

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.