CAJ | 학술논문

目的：骨髓基质细胞（bone marrow stromal cells，BMSCs）在调节颅骨锁骨发育不全（cleidocranial dysplasia，CCD）患者骨结构中发挥关键作用，本研究通过与正常 BMSCs 比较分析，探讨 CCD 患者 BMSCs 的体外增殖、成骨分化、干性及衰老特征。方法：分离培养 CCD 患者及正常同龄人 BMSCs；甲基噻唑基四唑（MTT）法及流式细胞周期分析其增殖能力；成骨诱导后采用Western blot 及茜素红染色分析其成骨能力；检测多潜能转录因子及克隆形成，分析其干性能力；检测衰老调控关键基因 p16、p21表达及通过β-gal 衰老染色分析其衰老特征。结果：与正常 BMSCs 相比，BMSCs-CCD 增殖活性低，且细胞周期中处于 S期、G2的细胞比例较低；两组细胞成骨诱导1、3、7 d 后，BMSCs-CCD 组中 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、骨桥蛋白（Osteopontin，Opn）表达水平较正常组低；成骨诱导14 d 后，BMSCs-CCD组形成的钙化结节较正常组少；BMSCs-CCD 组中多潜能转录因子 Oct4、Nanog、Sox2表达及克隆形成率均较正常组低；相反， BMSCs-CCD 组中 p16、p21等衰老标记分子表达及衰老细胞阳性染色比例均较正常组高。结论：同正常 BMSCs 比较，CCD 患者 BMSCs 的增殖能力、成骨能力、干性强度均较差，且更易衰老。这些特征可能是 CCD 患者易发骨质疏松及骨折的生物学机制之一。
목적：골수기질세포（bone marrow stromal cells，BMSCs）재조절로골쇄골발육불전（cleidocranial dysplasia，CCD）환자골결구중발휘관건작용，본연구통과여정상 BMSCs 비교분석，탐토 CCD 환자 BMSCs 적체외증식、성골분화、간성급쇠로특정。방법：분리배양 CCD 환자급정상동령인 BMSCs；갑기새서기사서（MTT）법급류식세포주기분석기증식능력；성골유도후채용Western blot 급천소홍염색분석기성골능력；검측다잠능전록인자급극륭형성，분석기간성능력；검측쇠로조공관건기인 p16、p21표체급통과β-gal 쇠로염색분석기쇠로특정。결과：여정상 BMSCs 상비，BMSCs-CCD 증식활성저，차세포주기중처우 S기、G2적세포비례교저；량조세포성골유도1、3、7 d 후，BMSCs-CCD 조중 Runt 상관전록인자2（Runt-related transcription factor 2，Runx2）、성골세포특이성전록인자 Osterix、골교단백（Osteopontin，Opn）표체수평교정상조저；성골유도14 d 후，BMSCs-CCD조형성적개화결절교정상조소；BMSCs-CCD 조중다잠능전록인자 Oct4、Nanog、Sox2표체급극륭형성솔균교정상조저；상반， BMSCs-CCD 조중 p16、p21등쇠로표기분자표체급쇠로세포양성염색비례균교정상조고。결론：동정상 BMSCs 비교，CCD 환자 BMSCs 적증식능력、성골능력、간성강도균교차，차경역쇠로。저사특정가능시 CCD 환자역발골질소송급골절적생물학궤제지일。
Objective:To study the in vitro biologic characteristics of bone marrow stromal cells with cleidocranial dysplasia (BM-SCs-CCD),including osteogenesis,proliferation ability,stemness and senescence.Methods:MTT and cell cycle detection for prolifera-tion ability,Western blot and alizarin red staining for osteogenesis ability,Western blot and clony-formation for stemness ability,Western blot and senescence staining for senescence characteristics.Results:The optical density value was lower in BMSCs-CCD in MTT experi-ment and lower BMSCs-CCD were under S or G2 stage while more under G1 stage in cell cycle detection.BMSCs-CCD expressed lower levels of Runx2,Opn,Osterix and weaker alizarin red staining.Moreover,BMSCs-CCD also exhibited weaker stemness associate proteins (Oct4,Nanog,and Sox2 )and clony-formation rates.However,stronger senescence associated proteins (p16,p21 )and more senes-cenced cells were found in BMSCs-CCD.Conclusions:BMSCs-CCD exhibited weaker proliferation ability,osteogenesis and stemness while stronger senescence ability as compared to BMSCs.These results are helpful for us to understand pathological mechanism of CCD bone disease.