摘要
目的:研究山羊乳牙牙髓干细胞在镁黄长石表面黏附、增殖和成骨分化特性。方法:采用酶消法培养山羊乳牙牙髓干细胞。在生长培养液条件下,应用荧光显微镜观察细胞在材料表面的黏附情况,以CCK-8法检测细胞在材料表面的增殖特性;在矿化培养液条件下,以pNPP法检测细胞在材料表面于1、4、7 d碱性磷酸酶(ALP)活性,并通过实时定量PCR检测4、7 d时成骨标志基因ALP、COL I、OPN和OCN的表达。采用SPSS 13.0软件包对数据进行统计学分析。结果:与β-TCP相比,山羊乳牙牙髓干细胞在镁黄长石表面伸展更开,在第4天起增长明显加快,ALP活性在4、7 d更高,镁黄长石组的基因表达水平明显上调,其中OCN表达量在第7天显著增加(P<0.01)。结论:山羊乳牙牙髓干细胞在镁黄长石表面能够良好黏附和增殖。与β-TCP相比,镁黄长石能够促进山羊乳牙牙髓干细胞向成骨方向分化。
PURPOSE: To explore the characters of attachment, proliferation and osteogenic differentiation of stem cells from goat deciduous teeth (SGDs) cultured on akermanite ceramic. METHODS: SGDs were collected with enzyme digestion, and cultured. Under culture condition in growth medium, actin filament labeling and CCK-8 analysis were used to reveal attachment and proliferation. Expression level of alkaline phosphatase (ALP) and osteogenic differentiation genes (ALP, COL I, OPN and OCN) were examined by pNPP method and real-time PCR. The data was analyzed using SPSS 13.0 software package. RESULTS: Compared with β-TCP, SGDs seeded on akermanite ceramic not only spread out better, but also grew faster since day 4. In osteogenic medium, ceils of akermanite group had stronger ALP activity at day 4 and 7. The gene expression of akermanite was more than the β-TCP group. The gene OCN expression level increased significantly at day 7 (P〈0.01). CONCLUSIONS: SGDs can attach and grow well on akermanite ceramic. Compared with β-TCP, akermanite ceramic shows an advantage in osteogenic differentiation of SGDs. Supported by Research Fund of Science and Technology Commission of Shanghai Municipality (09JC 1411700) and Research Fund of Shanghai Jiao Tong University School of Medicine (09XJ21030).
出处
《上海口腔医学》
CAS
CSCD
北大核心
2013年第2期146-150,共5页
Shanghai Journal of Stomatology
基金
上海市科学技术委员会资助项目(09JC1411700)
上海交通大学医学院基金项目(09XJ21030)~~
关键词
镁黄长石
山羊
乳牙牙髓干细胞
骨组织工程
成骨分化
Akermanite
Goats
Stem cells from goat deciduous teeth(SGDs)
Bone tissue engineering
Osteogenic differentiiation