2010 Volume 53 Issue 2 Pages 214-221
Periodontal diseases such as periodontitis represent a serious clinical challenge. The final goal in the treatment of periodontal diseases is the regeneration of periodontal tissue. Recent developments in tissue-engineering technology has led to the ex vivo cultivation of periodontal ligament (PDL) structures. Some studies have demonstrated that PDL-derived cells could regenerate periodontal tissues, and indicated that PDL cells and scaffolds have key roles in periodontal regeneration. However, there has been no report on the use of amniotic membrane (AM) as a scaffold for PDL-derived cell sheets. AM has high biocompatibility as well as anti-scarring and anti-infection properties. In the current study, we immunohistochemically evaluated PDL-derived cells cultured on AM. All experimental procedures introduced here were approved by the Kyoto Prefectural University of Medicine Ethics Committee. AM was obtained from women undergoing caesarean section. PDL tissue was obtained from a healthy human maxillary third molar that had erupted. The PDL tissue was minced and then cultured in a culture medium as explants. After 3rd to 4th passages, the PDL-derived cells were first cultured on an AM carrier and then evaluated immunohistochemically for distribution of Ki-67, vimentin, desmoplakin, and zonula occludens protein-1 (ZO-1), using an indirect fluorescent antibody method. PDL-derived cells showed a monolayered structure after 2 weeks in culture on AM. Immunohistochemistry revealed that PDL-derived cells cultured on AM expressed Ki-67, vimentin, desmoplakin, and ZO-1. These findings indicated that the cultivation of PDL-derived cells using AM enabled PDL-like differentiation. In addition, those cells showed desmosomes and tight junction-mediated cell-to-cell adhesion. We concluded that AM may represent a suitable scaffold for culturing PDL-derived cells, and that PDL-derived cells cultured on AM show sheet formation.
