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

Abstract EN

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.