CAJ | 학술논문

目的探讨机械牵张应力对体外培养的颈椎后纵韧带骨化症（OPLL）患者颈椎韧带成纤维细胞的影响。方法对2012年1月至2013年12月 OPLL 与颈椎外伤但无后纵韧带骨化（非OPLL）患者（各15例）行前路颈椎手术治疗，术中取韧带标本。采用组织块培养法进行细胞体外培养，免疫细胞化学及免疫荧光技术检测胞质波形蛋白。采用 Flexercell 4000细胞加载培养系统分别对两组患者第3代细胞进行机械牵张应力加载，逆转录-聚合酶链式反应（RT-PCR）方法检测两组成纤维细胞应力刺激前及刺激后12、24 h 成骨特异性指标骨钙素、碱性磷酸酶与Ⅰ型胶原 mRNA 表达。结果免疫细胞化学及免疫荧光检测显示胞质波形蛋白呈阳性表达。OPLL组后纵韧带成纤维细胞经机械牵张应力刺激12 h后，骨钙素、碱性磷酸酶及Ⅰ型胶原 mRNA 表达明显升高，应力刺激前后差异具有统计学意义；而非OPLL组应力刺激前后骨钙素、碱性磷酸酶及Ⅰ型胶原 mRNA 表达无明显变化。结论机械牵张应力可促使OPLL患者后纵韧带成纤维细胞骨钙素、碱性磷酸酶及Ⅰ型胶原mRNA表达增加，促进其骨化，其在OPLL进展中发挥重要作用。
목적탐토궤계견장응력대체외배양적경추후종인대골화증（OPLL）환자경추인대성섬유세포적영향。방법대2012년1월지2013년12월 OPLL 여경추외상단무후종인대골화（비OPLL）환자（각15례）행전로경추수술치료，술중취인대표본。채용조직괴배양법진행세포체외배양，면역세포화학급면역형광기술검측포질파형단백。채용 Flexercell 4000세포가재배양계통분별대량조환자제3대세포진행궤계견장응력가재，역전록-취합매련식반응（RT-PCR）방법검측량조성섬유세포응력자격전급자격후12、24 h 성골특이성지표골개소、감성린산매여Ⅰ형효원 mRNA 표체。결과면역세포화학급면역형광검측현시포질파형단백정양성표체。OPLL조후종인대성섬유세포경궤계견장응력자격12 h후，골개소、감성린산매급Ⅰ형효원 mRNA 표체명현승고，응력자격전후차이구유통계학의의；이비OPLL조응력자격전후골개소、감성린산매급Ⅰ형효원 mRNA 표체무명현변화。결론궤계견장응력가촉사OPLL환자후종인대성섬유세포골개소、감성린산매급Ⅰ형효원mRNA표체증가，촉진기골화，기재OPLL진전중발휘중요작용。
Objective To investigate the effect of mechanical stress on fibroblasts derived from ossification of the posterior longitudinal ligament.Methods From January 2012 to December 2013,15 patients with ossification of the posterior longitudinal ligament (OPLL)and 15 non-OPLL patients underwent anterior cervical decompression. The posterior longitudinal ligaments were collected intraoperatively,and then sent to the laboratory and underwent vitro cell culture by tissue fragment.Vimentin of cultured cells was measured with immunocytochemistry and immunofluorescence.The Flexercell 4000 Tension Plus system was used to stretch the third passage cells cultured from OPLL and non-OPLL patients.We detected the mRNA expression changes of osteocalcin (OCN),alkaline phosphatase (ALP)and type Ⅰ collagen (COLⅠ)by reverse transcription-polymerase chain reaction (RT-PCR)in the static condition at 12 h and 24 h after mechanical stress application.Results Cultured cells stained positively for vimentin by immunocytochemistry and immunofluorescence.The mRNA expressions of OCN,ALP and COLⅠsignificantly increased at 12 h after mechanical stress application in OPLL cells.However,there were no significant changes in the non-OPLL cells.Conclusion The mRNA expressions of OCN,ALP and COLⅠin OPLL cells could be significantly up-regulated by mechanical stress.Mechanical stress plays an important role in the progression of OPLL.