The effect of Emdogain on the growth and differentiation of rat bone marrow cells.
OBJECTIVE
The major extracellular matrix (ECM) proteins in developing enamel can induce and maintain the formation and mineralization of other skeletal hard tissue, such as bone. Therefore, dental matrix proteins are ideal therapeutic agents when direct formation of functional bone is required for a successful clinical outcome. Emdogain (EMD) consists of enamel matrix proteins which are known to stimulate bone formation. However, only a few studies in the literature have reported the effect of EMD on osteoblast-like cells in vitro.


METHODS
In this study, rat bone marrow cells, obtained from the femora of Wistar rats, were precultured for 7 d in osteogenic medium. Then, the cells were harvested and seeded in 24-well plates at a concentration of 20,000 cells/well. The wells were either precoated with 100 microg/ml EMD, or left uncoated. The seeded cells were cultured in osteogenic medium for 32 d and analysed for cell attachment (by using the Live and Dead assay), cell growth (by determining DNA content) and cell differentiation (by measuring alkaline phosphatase activity and calcium content, and by using scanning electron microscopy and the reverse transcription-polymerase chain reaction).


RESULTS
The results showed that at the 4-h time point of the experiment, more cells were attached to EMD-negative wells, but this effect was no longer apparent at 24 h. DNA analysis revealed that both groups showed a similar linear trend of cell growth. No differences in alkaline phosphatase activity or calcium content were observed, and no differences in gene expression (osteocalcin, alkaline phosphatase and collagen type I) were found between the groups.


CONCLUSIONS
Based on our results, we conclude that EMD had no significant effect on the cell growth and differentiation of rat bone marrow cells.
van den Dolder
J
J
Department of Periodontology and Biomaterials, Radboud University Nijmegen Medical Center, Nijmegen, the Netherlands. j.vandendolder@dent.umcn.nl
Vloon
A P G
AP
Jansen
J A
JA
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D001854:Bone Marrow Cells / Q000187:drug effects*; D002118:Calcium / Q000032:analysis; D002448:Cell Adhesion / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis; D003746:Dental Enamel Proteins / Q000494:pharmacology*; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods; D051381:Rats; D017208:Rats, Wistar
D003746:Dental Enamel Proteins; C109069:enamel matrix proteins; D004247:DNA; D000469:Alkaline Phosphatase; D002118:Calcium