Effects of enamel matrix derivative on proliferation/viability, migration, and expression of angiogenic factor and adhesion molecules in endothelial cells in vitro.
BACKGROUND
The aim of this study was to test in vitro the effect of enamel matrix derivative (EMD) on the proliferation/viability, migration, and expression of angiogenic factor and adhesion molecules in human umbilical vein endothelial cells (HUVECs). To date, discussions on angiogenic effects of EMD are rather controversial.


METHODS
The effect of EMD on the proliferation/viability of HUVECs after 24 hours was measured using 3,4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT) assay and direct cell counting. Cell migration was observed in an especially adapted in vitro monolayer wound-healing model. The expression of angiogenic factor angiopoietin-2 (ang-2) and adhesion molecules intercellular adhesion molecule (ICAM)-1 and vascular endothelium-selectin (E-selectin) was quantified with real-time polymerase chain reaction (PCR).


RESULTS
The proliferation/viability of HUVECs measured in MTT assay was stimulated by 0.1 microg/ml EMD and inhibited by higher doses (50 to 100 microg/ml), but the total number of cells was not affected. Cell migration in the wound-healing assay was promoted by EMD at doses of 0.1 to 50 microg/ml and inhibited at 100 microg/ml. The highest expression level of all three tested genes (ICAM-1, E-selectin, and ang-2) was observed at 50 microg/ml EMD.


CONCLUSIONS
The results of the present in vitro study show the potential influence of EMD on the angiogenic activity of HUVECs, which may play an important role in periodontal tissue regeneration and wound healing.
Department of Periodontology, Bernhard Gottlieb University Clinic of Dentistry, Vienna, Austria.
Kristina
K
Bertl
Na
N
An
Corinna
C
Bruckmann
Michel
M
Dard
Oleh
O
Andrukhov
Michael
M
Matejka
Xiaohui
X
Rausch-Fan
D043925:Angiogenesis Inducing Agents / Q000032:analysis; D042702:Angiopoietin-2 / Q000032:analysis; D002452:Cell Count; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004396:Coloring Agents; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D003746:Dental Enamel Proteins / Q000494:pharmacology*; D019040:E-Selectin / Q000187:drug effects; D042783:Endothelial Cells / Q000187:drug effects*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000187:drug effects; D013778:Tetrazolium Salts; D013844:Thiazoles; D013997:Time Factors; D053773:Transforming Growth Factor beta1 / Q000032:analysis; D014471:Umbilical Veins
D043925:Angiogenesis Inducing Agents; D042702:Angiopoietin-2; D004396:Coloring Agents; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D003746:Dental Enamel Proteins; D019040:E-Selectin; D013778:Tetrazolium Salts; D013844:Thiazoles; D053773:Transforming Growth Factor beta1; C109069:enamel matrix proteins; D018799:Intercellular Adhesion Molecule-1; C022616:thiazolyl blue