CAJ | 학술논문

目的探讨人牙髓细胞在体外传代过程中的生物学行为变化,为其在口腔材料生物相容性及牙髓损伤修复等研究中的应用提供依据.方法选取第4、8、10和15代人牙髓细胞,从细胞形态及增殖能力、碱性磷酸酶活性、老化相关β-半乳糖苷酶的表达及p53基因的表达对牙髓细胞进行评价.结果人牙髓细胞培养至第15代时,胞质增大、胞体扁平,增殖缓慢,第7天时A值仅为0.124±0.017,显著低于第4、8代(A值分别为0.391±0.030、0.317±0.049)(F=126.465,P =0.000);第15代细胞的碱性磷酸酶活性[(21.96±4.03) U/g蛋白]显著低于第4、8、10代细胞[分别为(85.67 ±4.64)、(84.22 ±2.99)、(29.64±3.53) U/g蛋白](P=0.000),并出现了大量β-半乳糖苷酶染色阳性细胞,p53基因表达显著上升,其相对表达量为(2.3±0.1),与第4代(设为1.0)比较差异有统计学意义(t=18.622,P=0.000).结论人牙髓细胞在传代培养过程中细胞体积增大、代谢酶指标下降、细胞老化相关酶及p53基因表达升高.
목적 탐토인아수세포재체외전대과정중적생물학행위변화,위기재구강재료생물상용성급아수손상수복등연구중적응용제공의거.방법 선취제4、8、10화15대인아수세포,종세포형태급증식능력、감성린산매활성、노화상관β-반유당감매적표체급p53기인적표체대아수세포진행평개.결과 인아수세포배양지제15대시,포질증대、포체편평,증식완만,제7천시A치부위0.124±0.017,현저저우제4、8대(A치분별위0.391±0.030、0.317±0.049)(F=126.465,P =0.000);제15대세포적감성린산매활성[(21.96±4.03) U/g단백]현저저우제4、8、10대세포[분별위(85.67 ±4.64)、(84.22 ±2.99)、(29.64±3.53) U/g단백](P=0.000),병출현료대량β-반유당감매염색양성세포,p53기인표체현저상승,기상대표체량위(2.3±0.1),여제4대(설위1.0)비교차이유통계학의의(t=18.622,P=0.000).결론 인아수세포재전대배양과정중세포체적증대、대사매지표하강、세포노화상관매급p53기인표체승고.
Objective To investigate biological behavior changes of human dental pulp cells during cell passaging,so as to provide a basis in the repair of dental pulp injury studies.Methods Human dental pulp cells at the 4 th,8 th,10 th and 15 th passage were investigated,including cells morphology,proliferation ability using methyl thiazolyl tetrazolium (MTT) assay,the alkaline phosphatase (ALP)expression and its activity,senescence-associated β-galactosidase (SA-β-Gal) expression and p53 gene expression.Results Cells alteration occurred at the 15 th passage.Cells morphology displayed flatter and larger plasma area.Cells proliferation ability decreased.The A value(0.124 ±0.017) at the 7 th day was significantly lower (F =126.465,P =0.000).ALP expression and activity significantly decreased [(21.96 ±4.03) U/g protein].More SA-β-Gal stain positive cells appeared.The p53 gene expression level was 2.3 times high compared with cells at the 4 th passage,the difference was statistically significant (t =18.622,P =0.000).Conclusions During cell passaging,human dental pulp cells exhibited senescence features in cellular level,enzymatic level and genic level separately.