Enamel matrix derivative stimulates osteogenesis- and chondrogenesis-related transcription factors in C3H10T1/2 cells.
Our purpose was to determine how enamel matrix derivative (EMD) affects the expression of osteogenesis- and chondrogenesis-related transcription factors in undifferentiated mesenchymal cells. C3H10T1/2 cell line, a typical pluripotential mesenchymal cell line, was cultured with or without EMD for up to 7 d. Expression of mRNAs encoding osteogenesis- and chondrogenesis-related transcription factors (Runx2, Osterix, AJ18, Dlx5, Msx2, Sox5, Sox9 and Zfp60) was measured using real-time polymerase chain reaction. Runx2 and Sox9 protein expression and the presence of bone morphogenetic protein (BMP)-6-like molecules in EMD were determined by Western blotting. EMD substantially increased mRNA levels of osteogenesis- and chondrogenesis-related transcription factors. EMD also induced Runx2 and Sox9 protein expression. Western blotting analysis of EMD using anti-BMP-6 antibody revealed immunoreactive bands corresponding to about 14 kDa and 60 kDa. These results suggest that EMD stimulates osteogenesis- and chondrogenesis-related transcription factors, and these activities may be mediated, at least in part, by BMP-6 in EMD.
Narukawa
Masataka
M
Department of Periodontology, Nihon University School of Dentistry, Tokyo101-8310, Japan.
Suzuki
Naoto
N
Takayama
Tadahiro
T
Yamashita
Yasuhiko
Y
Otsuka
Kichibee
K
Ito
Koichi
K
D000818:Animals; D055418:Bone Morphogenetic Protein 6; D019485:Bone Morphogenetic Proteins / Q000032:analysis; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000235:genetics*; D024043:Collagen Type II / Q000235:genetics; D003746:Dental Enamel Proteins / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D051379:Mice; D015675:Osteocalcin / Q000235:genetics; D010012:Osteogenesis / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis
C525817:Bmp6 protein, mouse; D055418:Bone Morphogenetic Protein 6; D019485:Bone Morphogenetic Proteins; D024043:Collagen Type II; D003746:Dental Enamel Proteins; D012333:RNA, Messenger; C109069:enamel matrix proteins; D015675:Osteocalcin