Development of tissue-engineered human periodontal ligament constructs with intrinsic angiogenic potential

Nobuhiro Nagai, Ayumi Hirakawa, Nao Otani, Masanobu Munekata

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)


One approach to treat periodontal diseases is grafting of tissue-engineered periodontal ligaments. Therefore, periodontal ligaments were constructed by layering cell sheets. A cell sheet was prepared by enzymatic digestion of salmon collagen gel on which human periodontal ligament fibroblasts (HPLFs) were co-cultured with or without human umbilical vein endothelial cells (HUVECs). Three cell sheets were layered and then cultured in angiogenic media, in which the HUVECs were found to form capillary-like structures when co-cultured on the HPLFs. The layered HPLFs sheets with HUVEC co-culture (PL-EC construct) demonstrated longer survival, higher alkaline phosphatase activities and lower osteocalcin production than layered HPLFs sheets without HUVEC co-culture (PL construct). Hematoxylin-eosin and Masson's trichrome staining of histological sections showed that cell density, mass and extracellular matrix deposition of the PL-EC construct were higher than those of the PL construct. Furthermore, CD31 immunostaining revealed the formation of capillary-like structures throughout the PL-EC construct. In conclusion, we successfully developed tissue-engineered periodontal ligament constructs with intrinsic angiogenic potential using cell sheet engineering and HUVEC co-culture.

Original languageEnglish
Pages (from-to)303-312
Number of pages10
JournalCells Tissues Organs
Issue number6
Publication statusPublished - 2009 Nov
Externally publishedYes


  • Angiogenesis
  • Cell culture
  • Cell sheet
  • Periodontal tissues
  • Salmon collagen
  • Tissue engineering

ASJC Scopus subject areas

  • Anatomy
  • Histology


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