Odontoblast marker gene expression is enhanced by a CC-chemokine family protein MIP-3alpha in human mesenchymal stem cells.
OBJECTIVE
Macrophage inflammatory protein-3 alpha (MIP-3alpha) is a major CC-chemokine family protein, which serves as a differentiation factor for mesenchymal cells, including osteoblasts and dental pulp cells. The purpose of this study was to investigate the influence of MIP-3alpha on human mesenchymal stem cell differentiation in vitro.


METHODS
Human mesenchymal stem cells were maintained in Dulbecco's modified Eagle's medium in the presence or absence of MIP-3alpha and the presence or absence of osteogenic factors (dexamethasone, beta-glycerophoshate and ascorbic acid). Alkaline phosphatase (ALP) activity was measured, and expression of odontoblast and osteoblast markers were examined by RT-PCR and Western blotting.


RESULTS
MIP-3alpha alone did not increase ALP activity, as compared to controls. The combination of MIP-3alpha and osteogenic factors increased ALP activity beyond increases observed with osteogenic factors alone. mRNA expression of the odontoblast marker dspp was only detectable when MIP-3alpha was added together with osteogenic factors at day 7 in three out of four samples. DSP protein level was increased only in the samples treated with both MIP-3alpha and osteogenic factors until day 5. In contrast, MIP-3alpha did not influence levels of the osteoblast markers CBFA1 or BSP.


CONCLUSIONS
The present study demonstrated that MIP-3alpha enhanced gene expression and protein levels of odontoblast-related genes, without affecting levels of the osteogenic proteins CBFA1 or BSP.
Iejima
D
D
Research and Development Center, Hitachi Medical Corporation, Kashiwa, Chiba 277-0804, Japan.
Sumita
Y
Y
Kagami
H
H
Ando
Y
Y
Ueda
M
M
D000328:Adult; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western / Q000379:methods; D002454:Cell Differentiation; D002478:Cells, Cultured; D019742:Chemokines, CC / Q000235:genetics / Q000502:physiology*; D003782:Dental Pulp / Q000166:cytology; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D019402:Macrophage Inflammatory Proteins / Q000235:genetics / Q000502:physiology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D009804:Odontoblasts / Q000166:cytology* / Q000502:physiology; D019707:Receptors, Chemokine; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012795:Sialoglycoproteins / Q000096:biosynthesis* / Q000235:genetics; D013223:Statistics as Topic
D015415:Biomarkers; D019742:Chemokines, CC; D019402:Macrophage Inflammatory Proteins; D019707:Receptors, Chemokine; D012795:Sialoglycoproteins