title: In vitro construction of scaffold-free bilayered tissue-engineered skin containing capillary networks.
abstract: Many types of skin substitutes have been constructed using exogenous materials. Angiogenesis is an important factor for tissue-engineered skin constructs. In this study, we constructed a scaffold-free bilayered tissue-engineered skin containing a capillary network. First, we cocultured dermal fibroblasts with dermal microvascular endothelial cells at a ratio of 2 : 1. A fibrous sheet was formed by the interactions between the fibroblasts and the endothelial cells, and capillary-like structures were observed after 20 days of coculture. Epithelial cells were then seeded on the fibrous sheet to assemble the bilayered tissue. HE staining showed that tissue-engineered skin exhibited a stratified epidermis after 7 days. Immunostaining showed that the epithelium promoted the formation of capillary-like structures. Transmission electron microscopy (TEM) analysis showed that the capillary-like structures were typical microblood vessels. ELISA demonstrated that vascularization was promoted by significant upregulation of vascularization associated growth factors due to interactions among the 3 types of cells in the bilayer, as compared to cocultures of fibroblast and endothelial cells and monocultures.
authors.affiliation: Department of Oral Histology and Pathology, School of Stomatology, The Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Liu
authors.forename: Hailang
authors.initials: H
authors.lastname: Luo
authors.forename: Xinwen
authors.initials: X
authors.lastname: Wang
authors.forename: Akimichi
authors.initials: A
authors.lastname: Takemura
authors.forename: Yi Ru
authors.initials: YR
authors.lastname: Fang
authors.forename: Yan
authors.initials: Y
authors.lastname: Jin
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Suwa
mesh_terms: D002196:Capillaries / Q000166:cytology; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000166:cytology; D000078404:Epidermal Cells; D004817:Epidermis / Q000254:growth & development; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D018919:Neovascularization, Physiologic; D017768:Skin, Artificial; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D014471:Umbilical Veins / Q000166:cytology