Development of tissue-engineered human periodontal ligament constructs with intrinsic angiogenic potential.
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.
Nagai
Nobuhiro
N
Creative Research Initiative 'Sousei', Hokkaido University, Sapporo, Japan. nagai491@yahoo.co.jp
Hirakawa
Ayumi
A
Otani
Nao
N
Munekata
Masanobu
M
D000818:Animals; D002450:Cell Communication; D002454:Cell Differentiation; D049109:Cell Proliferation; D018920:Coculture Techniques; D003094:Collagen; D003470:Culture Media; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology; D005109:Extracellular Matrix / Q000502:physiology; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D006801:Humans; D018919:Neovascularization, Physiologic; D010513:Periodontal Ligament / Q000098:blood supply / Q000166:cytology / Q000502:physiology*; D012038:Regeneration; D023822:Tissue Engineering / Q000379:methods*
D003470:Culture Media; D003094:Collagen