| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2002: ¥1,200,000 (Direct Cost: ¥1,200,000)
|
|---|
| Research Abstract |
Many mesenchyme cells exist in the dental pulp tissue, these cells may be differentiated to capable of hard tissue formation by various stimuli. The purpose of this study is the mechanisms that the mechanisms of hard tissue formation on dental pulp is clarified. The cells were prepared by outgrowth from normal dental pulp obtained from the third molar extracted under aseptic conditions from a 20-year-old patient during orthodontic treatment. The patient gave informed consent before providing the sample. After the dental pulp was extracted, the tissue was minced, placed on a 35-mm^2 tissue-culture dish and then covered with a sterile glass coverslip.
ALP activity of dental pulp cells which were treated with 50 μM ascorbic acid, 10 mMβ-glycerophosphate, 100 nM dexamethasone after treated with 1 mM 5-Azacytidine was more strongly, than none treated 5-Azacytidine. It seemed that the dental pulp cells were de-differentiated by 5-Azacytidine. It may be possible that a number of dental pulp cells were induced cells with activity of mineralization. It seems that by 5-Azacytidine made de-metylation to the cells and many cells were differentiated. It is found that many gene when dental pulp cells were stimulated to hard tissue formation.
HDP cells were incubated with/without 50 mg/ml EMD and mRNA were extracted. The fluorescent labeled samples were hybridized on a human cDNA microarray (Clontech, 1,100 genes ), and the fluorescence intensity of each gene was measured using an array scanner. From the analytical results of gene expression fluctuation of pulp cell with EMD, many genes increased their mRNA levels including serum iducible kinase, cycline dependent kinase 2 and IGF-1. Identified genes found in this study may involved in cell proliferation and calcification process. Gene expression profiling using cDNA microarray shoud be useful understanding the biological effect to HDP cells.
|
