摘要
目的：应用硅酸钙（CaSiO3，CS）生物陶瓷作用于人牙髓细胞（human dental pulp cells，hDPCs），研究其对hDPCs增殖及向成牙本质细胞方向分化的影响。方法：从年轻健康患者（18～20岁）拔除的智齿或前磨牙牙髓组织中获取hDPCs进行培养。将质量浓度为0．2g／mL的CS浸提液按1／2、1，4、1／8、1／16、1／32、1／64和1／128倍比稀释后作用于hDPCs。MTY实验检测不同质量浓度CS浸提液对hDPCs增殖的影响，进而筛选出最佳浓度。以最佳质量浓度（1／64倍比稀释）CS浸提液培养hDPCs2、4d后，Real．TimePCR检测以下hDPCs成牙本质相关基因的表达：牙本质涎磷蛋白（dentin sialophosphopmtein，DSPP）、牙本质基质蛋白－1（dentin matrix protein1，DMP．1），I型胶原（collagen tvpeI，COL-I）、骨钙蛋白（osteocalcin，OCN）、骨桥蛋白（osteopontin，OPN）；培养7、14d后碱性磷酸酶（alkaline phosphatase，ALP）染色及半定量检测ALP活性。结果：最佳质量浓度（1／64倍比稀释）cs浸提液能够促进hDPCs的增殖，对牙髓细胞DSPP、DMP-1、COL-1、OPN等成牙本质相关基因的表达有较为明显的促进作用。ALP染色及半定量分析显示该浓度的CS浸提液能够提高hDPCs分泌ALP的活性。结论：最佳质量浓度（1／64倍比稀释）CS浸提液能够明显促进hDPCs的增殖，提高成牙本质相关基因的表达，进而促进hDPCs向成牙本质细胞方向分化，为后期hDPCs结合CS支架材料进行牙本质再生的研究打下基础。
Objective： This paper was aimed to investigate the influence of calcium silicate （CS）ceramie on osteoblastic and odontoblastic differentiation of human dental pulp cells （hDPCs） in vitro. Methods： The hDPCs were derived from healthy third molars or premolars that had been extracted from young healthy patients （18-20 years old）. 0.2g/mL CS extract was diluted to 1/2, 1/4, 1/8, 1/16, 1/32, 1/64 and 1/128 concentration. The hDPCs were incubated in the medium that contain different CS extract dilution, hDPCs proliferation was studied by MTT assay on days 1, 2, and 4 to determine the optimal CS extract dilution. The odontogenesis related genes including dentin sialophosphoprotein （DSPP）, dentin matrix protein 1 （DMP- 1）, collagen type I （COL-1）, osteopontin （OPN）,osteocalein （OCN） were tested by Real-time PCR on days 2 and 4. ALP activity at time-point day 7 and day 14 was determined calorimetrically with a substrate solution. Results： MTT assay showed the proliferation of hDPCs were significantly increased when incubated with CS in a concentration of 1/64 dilution. Theexpression of odontogenesis related gene DSPP, DMP-1, COL-1 and OPN were enhanced in the optimal CS group. ALP staining and semi-quantitative analysis indicated that CS enhanced the ALP activity of hDPCs. Conclusion： Results show that the CS bioceramics could enhance the proliferation and promote odontogenesis differentiation of hDPCs in vitro. The combination of CS scaffolds and human dental pulp cells for dentin regeneration showed a promising prospect.
出处
《口腔颌面外科杂志》
CAS
2014年第1期4-9,共6页
Journal of Oral and Maxillofacial Surgery
基金
国家自然科学基金项目(81170939)
上海科学技术委员会项目资助(12nm0501600)
关键词
牙髓细胞
硅酸钙生物陶瓷
增殖
分化
dental pulp cells
calcium silicate bioceramics
proliferation
differentiation
