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

作　　者：戈杰[1,2] 张娟[1,2] 郭松松[1,2] 傅瑜[1,2] 江宏兵[1,2] 

机构地区：[1]南京医科大学口腔疾病研究江苏省重点实验室,江苏南京210029 [2]南京医科大学附属口腔医院口腔颌面外科,江苏南京210029

出　　处：《口腔生物医学》2014年第4期169-173,共5页Oral Biomedicine

基　　金：国家自然科学基金(81070810;81470723);江苏高校优势学科建设工程资助项目(2014-37)

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Brd U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Brd U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。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：DF-Cs-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.

关 键 词：颅骨锁骨发育不全 牙囊细胞 增殖 成骨 破骨 

分 类 号：R816.1[医药卫生—放射医学]