颅骨锁骨发育不全患者牙囊细胞的体外生物学特征

›› 2014, Vol. 5 ›› Issue (4): 169-173.

• 论著 • 下一篇

颅骨锁骨发育不全患者牙囊细胞的体外生物学特征

戈杰,张娟,郭松松,傅瑜,江宏兵

江苏省南京医科大学附属口腔医院口腔颌面外科

收稿日期:2014-08-29 修回日期:2014-12-10 出版日期:2014-12-25 发布日期:2015-01-06
通讯作者: 江宏兵 E-mail:jhbtooth@163.com
基金资助:
国家自然科学基金资助项目;江苏高校优势学科建设工程资助项目

Invitro biologic characteristics of dental follicle cells with cleidocranial dysplasia

Received:2014-08-29 Revised:2014-12-10 Online:2014-12-25 Published:2015-01-06

摘要/Abstract

摘要： 目的：在成功分离培养正常同龄人牙囊细胞（dental follicle cells，DFCs）与颅骨锁骨发育不全（cleidocranial dysplasia，CCD）患者牙囊细胞（DFCs-CCD）的基础上，研究其一般体外生物学特征，包括增殖、克隆、成骨及破骨能力。方法：采用BrdU细胞增殖实验与倍增法研究两种来源DFCs增殖能力；用结晶紫染液染色培养12 d的两种DFCs，分析其克隆形成能力；并用成骨诱导液诱导两种DFCs成骨，用Western blot和茜素红染色法分析成骨能力，用实时定量PCR方法研究其破骨基因表达差异。结果：DFCs-CCD的BrdU阳性率高于DFCs，同时DFCs的群体倍增时间为（1.834±0.093）d，而DFCs-CCD的则为（1.394±0.028）d，差异具有统计学意义（P<0.05）；DFCs-CCD的克隆集落多于DFCs，但Runt相关转录因子2（Runt-related transcription factor 2，Runx2）、成骨细胞特异性转录因子Osterix、骨钙素（osteocalcin，Ocn）等成骨相关蛋白表达水平低，而其茜素红染色所示钙化结节同样较少；同时，DFCs-CCD高表达核因子-κB受体活化因子（receptor activator for nuclear factor-κB，RANK）和骨保护素（osteoprotegerin，OPG）等破骨相关基因（P<0.05），而核因子-κB受体活化因子配体（receptor activator for nuclear factor-κB ligand，RANKL）水平无统计学差异（P＞0.05）。结论：相比DFCs，DFCs-CCD具有更强的增殖、克隆能力和更弱的成骨能力，而破骨基因表达紊乱。

Abstract: Objective： To study the invitro biologic characteristics of dental follicle cells with cleidocranial dysplasia (DFCs-CCD) including proliferation ability, clone-forming efficiency, osteogenesis and osteoclastogenesis abilities. Methods： Brdu cell proliferation and population doubling time assays for cell proliferationo ability, crystal violet staining with cells cultured for 12 days for clone-forming efficiency, Western blot and alizarin red staining for osteogenesis ability and real-time PCR for osteoclastogenesis ability. Results： DFCs-CCD gave higher BrdU positive results. The population doubling time was (1.834±0.093) d of DFCs and (1.394±0.028) d of DFCs-CCD (P<0.05). DFCs-CCD presented more clones and weaker expression of Runx2, Osrerix, Ocn and alizarin red staining. DFCs-CCD also expressed stronger levels of OPG and RANK, but no statistical difference of RANKL(P＞0.05). Conclusions：DFCs-CCD exhibited stronger cell proliferation and clone-forming abilities, weaker osteogenesis and disorganized osteoclastogenesis abilities.

戈杰张娟郭松松傅瑜江宏兵. 颅骨锁骨发育不全患者牙囊细胞的体外生物学特征[J]. , 2014, 5(4): 169-173.

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颅骨锁骨发育不全患者牙囊细胞的体外生物学特征

Invitro biologic characteristics of dental follicle cells with cleidocranial dysplasia

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