硅酸钙生物陶瓷对牙髓细胞体外增殖分化的影响

doi:10.3969/j.issn.1005-4979.2014.01.002

《口腔颌面外科杂志》 ›› 2014, Vol. 24 ›› Issue (1): 4-. doi: 10.3969/j.issn.1005-4979.2014.01.002

硅酸钙生物陶瓷对牙髓细胞体外增殖分化的影响

张茂林¹，夏伦果¹，陈蕾²，张秀丽³，蒋欣泉³，张志愿¹

1. 上海交通大学医学院附属第九人民医院口腔颌面外科，上海200011；
2. 中国科学院上海硅酸盐研究所高性能陶瓷和超微结构国家重点实验室, 上海200050；
3. 上海交通大学医学院附属第九人民医院口腔修复科，口腔生物工程/再生医学实验室，上海 200011

出版日期:2014-02-28 发布日期:2014-06-27
通讯作者: 张志愿，教授. E-mail:zhzhy@omschina.org.cn
作者简介:张茂林（1986—）, 男，河北人，硕士研究生. E-mail:zml0312@163.com
基金资助:
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）

Calcium Silicate for the Mineralization of Human Dental Pulp Cells

ZHANG Mao-lin¹, XIA Lun-guo¹, CHEN-Lei², ZHANG-Xiu li³, JIANG Xin-quan³, ZHANG Zhi-yuan¹

1. Department of Oral and Maxillofacial Surgery, College of Stomatology, Ninth People′s Hospital, Shanghai Jiaotong University, School of Medicine, Shanghai 200011;2. State Key Laboratory of High Performance Ceramics and Superfine Microstructure, Shanghai Institute of Ceramics, Chinese Academy of Sciences, Shanghai 200050;3. Department of Prosthodontics,Ninth People′s Hospital, Shanghai Jiaotong University; Oral Bioengineering and Regenerative Medicine Lab,Shanghai 200011，China

Online:2014-02-28 Published:2014-06-27

摘要/Abstract

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0.2 g/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。MTT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hDPCs 2、4 d后，Real?鄄Time PCR检测以下hDPCs成牙本质相关基因的表达：牙本质涎磷蛋白（dentin sialophosphoprotein, DSPP）、牙本质基质蛋白?鄄1（dentin matrix protein 1, DMP?鄄1），Ⅰ型胶原（collagen typeⅠ, COL?鄄Ⅰ）、骨钙蛋白（osteocalcin, OCN）、骨桥蛋白（osteopontin, OPN）；培养7、14 d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?鄄1、COL?鄄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。

关键词: 牙髓细胞； , 硅酸钙生物陶瓷； , 增殖； , 分化

Abstract: Objective: This paper was aimed to investigate the influence of calcium silicate(CS)ceramic 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),osteocalcin (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. The expression 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.

Key words: dental pulp cells, calcium silicate bioceramics, proliferation, differentiation

中图分类号:

R782.2

张茂林1，夏伦果1，陈蕾2，张秀丽3，蒋欣泉3，张志愿1. 硅酸钙生物陶瓷对牙髓细胞体外增殖分化的影响[J]. 《口腔颌面外科杂志》, 2014, 24(1): 4-.

ZHANG Mao-lin1, XIA Lun-guo1, CHEN-Lei2, ZHANG-Xiu li3, JIANG Xin-quan3, ZHANG Zhi-yuan1. Calcium Silicate for the Mineralization of Human Dental Pulp Cells[J]. 《Journal of Oral and Maxillofacial Surgery》, 2014, 24(1): 4-.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2014/V24/I1/4

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