고농도의 포도당이 처리된 MC3T3-E1 세포에서 635 nm 광조사에 의한 골분화에 관한 연구
(주)코리아스칼라
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서지정보
ㆍ발행기관 : 대한구강악안면병리학회
ㆍ수록지정보 : 대한구강악안면병리학회지 / 35권 / 1호
ㆍ저자명 : 임원봉, 원재웅, 김지선, 김인애, 고영종, 권혁일, 김상우, 김서연, 김옥준, 최홍란
목차
I. INTRODUCTION
II. MATERIALS AND METHODS
1. Cell culture and chemicals
2. Light source and irradiation
3. Measurement of calcium deposition
4. Alizarin red S staining
5. Alkaline phosphatase(ALP) assay
6. Total RNA isolation and quantitative RT-PCR
7. Cell viability
8. Statistical analysis
III. RESULTS
1. Bone mineralization and ALP activity
2. mRNA expression of MC3T3-E1 differentiation-related genes
3. Determination of PMA concentration
4. Bone mineralization on PMA-treatedMC3T3-E1 cells
5. mRNA expression in PMA-treatedMC3T3-E1 cells
IV. DISCUSSION
V. REFERENCES
영어 초록
Diabetic patients tend to exhibit delayed bone formation and osteoblast differentiation, which results in osteopenia. Recently, numerous clinical reports suggest that 635-nm light irradiation improves bone regeneration and wound healing, and reduces pain in patients suffering from diabetes. The purpose of the present study was to test the hypothesis that 635-nm irradiation can influence bone formation by MC3T3-E1 osteoblasts cultured on high concentrations of glucose(25mmol/L D-glucose) in the presence or absence of phorbol 12-myristate 13-acetate(PMA), and to establish an in vitro pathological model of bone formation. The effect of 635-nm irradiation on bone formation was investigated using Alizarin Red S staining, and alkaline phosphatase enzyme activ ity and calcium deposition assays. In addition, gene expression of the o steogenic markers BMP-2, osterix and osteocalcin were assayed by RT-PCR. Calcium deposition by MC3T3-E1 cells was reduced in the presence of high concentrations of glucose or by PMA supplementation. However, 635-nm irradiation led to an increase in calcium deposition by MC3T3 cells, followed by increased bone mineralization. mRNA expression of BMP-2 and osterix at an early stage and of osteocalcin at a late stage was significantly upregulated by 635-nm irradiation in MC3T3-E1 cells supplemented with high concentrations of glucose. Irradiation at 635 nm increases bone mineralization in MC3T3-E1 cells cultured in vitro on high concentrations of glucose and alters osteogenic gene expression, which accelerates bone formation in hyperglycemic conditions.
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