The influence of enamel matrix derivative on the angiogenic activity of primary endothelial cells.
OBJECTIVE
Angiogenesis plays a crucial role in early wound healing and tissue regeneration. Although enamel matrix derivative (EMD) has demonstrated the potential to stimulate periodontal regeneration, the biological effects of EMD on angiogenesis and underlying mechanisms have not been fully elucidated. The aim of the present study was to examine the angiogenic effects of EMD in vitro.


METHODS
Human umbilical vein endothelial cells (HUVECs) were used to assess the effect of EMD on proliferation, survival, adhesion and migration. The effect of EMD on HUVEC angiogenesis was assessed by a three-dimensional sprouting assay. In order to understand the signalling mechanism of altered cell proliferation of HUVECs caused by EMD, the phosphorylation status of ERK1/2 and of the serine/threonine protein kinase Akt was analysed by western blot using phospho-specific antibodies.


RESULTS
The proliferation of HUVECs was stimulated by 50 μg/mL EMD, whereas higher concentrations (≥100 μg/mL) resulted in an increased apoptotic rate. The mitogenic response to EMD was associated with the activation of ERK1/2. Enamel matrix derivative did not affect cell adhesion, but all concentrations of EMD tested (0.1-250 μg/mL) promoted migration of HUVECs. Furthermore, EMD induced capillary-like sprout formation from HUVEC spheroids in a dose-dependent manner.


CONCLUSIONS
Our data indicate that EMD acts as a proangiogenic factor in vitro and, as such, might contribute to periodontal tissue regeneration by stimulation of vessel formation during wound healing.
Department of Operative Dentistry and Periodontology, University Medical Center, Johannes Gutenberg-University Mainz, Mainz, Germany. kasaj@gmx.de
A
A
Kasaj
J
J
Meister
K
K
Lehmann
S I
SI
Stratul
M
M
Schlee
J M
JM
Stein
B
B
Willershausen
M
M
Schmidt
D017209:Apoptosis; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003746:Dental Enamel Proteins / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects*; D004789:Enzyme Activation; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D010766:Phosphorylation; D061251:Primary Cell Culture; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D018874:Spheroids, Cellular / Q000187:drug effects; D014471:Umbilical Veins / Q000166:cytology; D014945:Wound Healing / Q000187:drug effects*
D003746:Dental Enamel Proteins; C109069:enamel matrix proteins; D051057:Proto-Oncogene Proteins c-akt; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3