Effects of enamel matrix derivative on proliferation and differentiation of primary osteoblasts.
OBJECTIVE
The aim of this study was to investigate the effects of enamel matrix derivative (EMD) on proliferation, protein synthesis, and mineralization in primary mouse osteoblasts.


METHODS
Osteoblasts were obtained from mouse calvaria by enzymatic digestion and grown in monolayer together with EMD (2-100 microg/ml). Metabolic activity and cell proliferation were determined by tetrazolium salt assay (MTT) and by 5-bromo-2'-deoxyuridine (BrdU) incorporation. For differentiation studies, a 3-dimensional organoid culture system was used. Osteoblastic differentiation was estimated by alkaline phosphatase (ALP) activity and calcium content. Collagen synthesis was assessed by [(3)H]-proline incorporation. Morphologic observations were made by electron microscopy.


RESULTS
EMD treatments increased metabolic cell activity and BrdU incorporation. In the organoid cultures, ALP activity and calcium accumulation were enhanced by EMD treatment, but [(3)H]-proline incorporation was not. Morphologically, an increased deposition of mineralized nodules was found.


CONCLUSIONS
EMD treatment enhanced cellular activities of primary osteoblasts and might support the regeneration of periodontal bony defects.
Department of Periodontology, Charité Berlin Germany. stefan.haegewald@charite.de
Stefan
S
Hägewald
Nicole
N
Pischon
Przemyslaw
P
Jawor
Jean-Pierre
JP
Bernimoulin
Bernd
B
Zimmermann
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D000963:Antimetabolites; D001973:Bromodeoxyuridine; D002118:Calcium / Q000032:analysis; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D003094:Collagen / Q000187:drug effects; D004396:Coloring Agents; D003746:Dental Enamel Proteins / Q000494:pharmacology*; D051379:Mice; D008854:Microscopy, Electron; D010006:Osteoblasts / Q000187:drug effects* / Q000378:metabolism; D013778:Tetrazolium Salts; D013844:Thiazoles
D000963:Antimetabolites; D004396:Coloring Agents; D003746:Dental Enamel Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; C109069:enamel matrix proteins; D003094:Collagen; D000469:Alkaline Phosphatase; C022616:thiazolyl blue; D001973:Bromodeoxyuridine; D002118:Calcium