Growth Factors and Biological Supports for Endothelial Cell Lining: In Vitro Study

Abstract

Endothelial cell covering over the vascular prosthesis luminal surface is a process that may require the presence of growth factors (GFs) and extracellular matrix supports. Endothelialization could be improved by combining both GFs and an extracellular matrix analog. In the present study, different biological substrates made of type I or IV collagens, gelatin, fibronectin, fibrin, laminin, chondroitin sulphate, heparan sulphate, heparin or hyaluronic acid were used to support endothelial cell culture. An endothelial cell growth supplement (ECGS) was incorporated in (group 1) or overlaid on (group 2) the substrates; or present in medium (group 3); or absent (group 4). GF binding assay using ¹²⁵I bFGF showed that more GF remained combined to the substrates in group 2 than those in group 1. Growth and morphology of human umbilical vein endothelial cells were sequentially analyzed in vitro for 8 days using DNA (nuclei counts) and F-actin labelings. Growth was relatively stable for the first 48 hours, later in groups 1, 2 and 4, cell death was observed on all the substrates except for fibronectin. Growth failure could be related to the degradation or inefficient release of ECGS. In group 3, growth increased and confluency was reached within 5–8 days on all the substrates except for gelatin and type I collagen. Confluent cells containing actin filaments were organized on glycoproteins and disorganized on glycosaminoglycans and fibrin. Despite that glycoproteins can enhance cell adhesion and lining pattern, GFs continually delivered in a fresh soluble form seem to be the appropriate condition to obtain an endothelial cell lining.