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BioMed Research International
Volume 2013 (2013), Article ID 561410, 8 pages
http://dx.doi.org/10.1155/2013/561410
Research Article

In Vitro Construction of Scaffold-Free Bilayered Tissue-Engineered Skin Containing Capillary Networks

1Department of Oral Histology and Pathology, School of Stomatology, The Fourth Military Medical University, Xi'an 710032, China
2Xi'an Institute of Tissue Engineering & Regenerative Medicine, Xi'an 710032, China
3Department of Periodontology & Oral Medicine, School of Stomatology, The Fourth Military Medical University, Xi'an 710032, China
4Department of Anatomy, Osaka Dental University, 8-1 Kuzuhahanazono-cho, Hirakata-shi, Osaka 573-1121, Japan
5Department of Dental Oriental Medicine, Osaka Dental University, 8-1 Kuzuhahanazono-cho, Hirakata-shi, Osaka 573-1121, Japan

Received 5 January 2013; Revised 21 February 2013; Accepted 22 February 2013

Academic Editor: Richard Tucker

Copyright © 2013 Yuan Liu et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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.