ABSTRACT
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 (5h) 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.
Subject(s)
Angiogenic Proteins/metabolism , Dental Pulp/injuries , Dental Pulp/metabolism , Neovascularization, Physiologic , Wound Healing/physiology , Adolescent , Cells, Cultured , Dose-Response Relationship, Drug , Enzyme-Linked Immunosorbent Assay/methods , Fibroblast Growth Factor 2/analysis , Fibroblast Growth Factor 2/metabolism , Humans , Methacrylates/pharmacology , Platelet-Derived Growth Factor/analysis , Platelet-Derived Growth Factor/metabolism , Vascular Endothelial Growth Factors/analysis , Vascular Endothelial Growth Factors/metabolismABSTRACT
AIM: To record the cytotoxicity of Resilon and Epiphany (Pentron clinical technologies, Wallingford, CT, USA) using a root model. METHODOLOGY: Thirty teeth with single roots were sectioned at the enamel-cementum junction, the root canals prepared and each root then sterilized before filling with the lateral condensation technique using one of three filling materials (n = 10 per group): Resilon and Epiphany, Sealite (Septodont, Pierre Rolland, Merignac, France) and gutta-percha, Roekoseal Automix (Coltène/Whaledent, Langenau, Germany) and gutta-percha. The roots were stored at 37 degrees C in an incubator to allow for setting of the root filling materials. The apices of the roots were dipped in 1 mL of MEM culture medium for 1, 2, 7 and 30 days renewing the medium every day. After 24 h contact between the medium and the filled roots, the medium was used to measure the cytotoxicity on mouse fibroblasts L 929 with the MTT assay that recorded the mitochondrial activity of the target cells. An additional test according to ISO 10993-5 standards was undertaken to compare Resilon and Epiphany. RESULTS: The root model showed no statistically significant differences between the sealers at 7 and 30 days (NS). Epiphany and Resilon were the most cytotoxic materials at 1 and 2 days (P < 0.001). Unlike Epiphany, Resilon was not cytotoxic when tested according to ISO 10993-5 standards. CONCLUSIONS: The cytotoxicity of Resilon + Epiphany, due mainly to Epiphany, decreased after 2 days to reach a level comparable with commonly used root canal sealers.
Subject(s)
Root Canal Filling Materials/toxicity , Analysis of Variance , Animals , Cattle , Confidence Intervals , Dental Cements/toxicity , Humans , Mice , Toxicity Tests , Zinc Oxide-Eugenol Cement/toxicityABSTRACT
After pulp amputation, complete pulp healing requires not only reparative dentin production but also fibroblast proliferation, nerve fiber growth, and neoangiogenesis. This study was designed to investigate the role of pulp fibroblasts in angiogenesis. Human pulp fibroblasts from third molars co-cultured with human umbilical vein endothelial cells induced the organization of endothelial cells and the formation of tubular structures corresponding to capillaries in vivo. The direct contact between both cells was not necessary to induce angiogenesis, and the observed effect was due to soluble factors. This was confirmed with neutralizing antibodies against FGF-2 and VEGF, which decreased the angiogenic effects of these soluble factors. Immunohistochemistry showed that both FGF-2 and VEGF were expressed in human dental pulp fibroblasts, and this expression increased after injury. These results suggest that the pulp fibroblasts secrete angiogenic factors, which are necessary for complete pulp healing, particularly at the pulp injury site.
