Quantification of angiogenic growth factors released by human dental cells after injury

Objective: Angiogenesis is a key step in the dental pulp healing sequence which involves the dentine bridge formation. In a previous work, we showed that dental pulp cells secrete soluble factors which interact with endothelial cells and affect the process of angiogenesis. The objective of this work was to quantify the angiogenic growth factors released by mechanically injured human dental pulp cells and the effect of 2-hydroxyethyl methacrylate (HEMA) on this secretion. Design: Pulp cells were prepared from immature third molars explants by the outgrowth method. Cell monolayers were either subjected to mechanical injuries or treated with increasing concentrations of HEMA. ELISA was used to quantify the secreted angiogenic growth factors in the culture media after different time periods of injury and after incubation with different concentrations of HEMA. Results: Pulp cells secreted significant levels of PDGF-AB, VEGF and FGF-2. The concentration of these factors increased shortly (5 h) after injury and returned to initial values after 1 day. HEMA treatment increased VEGF secretion but decreased that of FGF-2 in a dose-dependent manner while it did not affect PDGF-AB level. Conclusions: Dental pulp cells secrete angiogenic growth factors which play a pivotal role in angiogenesis which precedes the reparative dentine formation. PDGF-AB seems to play a major role because its level showed the highest increase in mechanically injured cells. The presence of HEMA affects both FGF-2 and VEGF levels and may partially explain the lack of dentine bridging after direct pulp capping with an adhesive system. (C) 2007 Elsevier Ltd. All rights reserved.