TITLE:
In Vitro Analysis of VEGF and HGF Production by Fibroblast in Cultured Dermal Substitute Combined with EGF-Incorporating Top Dressing
AUTHORS:
Emi Iijima, Daichi Daichi Toyoda, Akiko Yamamoto, Misato Kuroyanagi, Yoshimitsu Yoshimitsu Kuroyanagi
KEYWORDS:
Cultured Dermal Substitute; Fibroblast; Collagen; Hyaluronic Acid; Epidermal Growth Factor
JOURNAL NAME:
Open Journal of Regenerative Medicine,
Vol.3 No.1,
March
3,
2014
ABSTRACT: This study aimed to investigate the
potential of cultured dermal substitute (CDS) to release angiogenic growth
factors when laminated with a membrane containing epidermal growth factor (EGF)
as a top dressing. Membranes were prepared by air-drying a solution of
hyaluronic acid (HA) and collagen (Col) with or without EGF. Membranes were
designed to contain EGF at concentrations of 0, 0.1, 0.2 or 0.5 μg/cm2. CDS was
prepared by incorporating fibroblasts into a collagen gel combined with a
cross-linked HA spongy matrix, followed by culturing for 5 days. CDS was
designed to contain fibroblasts at a density of 2 × 105 (Group I) or 4 × 105 cells/cm2> (Group II). CDS was elevated at the interface between air and culture medium,
on the top of which each membrane was placed. This culture system was employed
as a wound surface model. Metabolic activity of the fibroblasts in the CDS
cultured for 7 days on a wound surface model was measured by MTT assay. The
amounts of vascular endothelial growth factor (VEGF) and hepatocyte growth
factor (HGF) after 7 days of cultivation were measured by using ELISA.
Membranes containing EGF ranging from 0.1 to 0.5 μg/cm2> facilitated production
of both VEGF and HGF, as compared with control membranes without EGF. However,
a membrane containing EGF at a concentration of 0.5 μg/cm2> failed to facilitate
fibroblast cytokine production in Group I. These results demonstrated that the
EGF-incorporating membrane was able to stimulate fibroblasts in the CDS to
synthesize an increased amount of VEGF and HGF in a dose-dependent manner.