CAJ | 학술논문

背景:众所周知,骨重建是骨组织中重要的生物学反应过程,其中成骨细胞与破骨细胞发挥了关键作用.但目前,关于骨重建中成骨与破骨细胞间信号传递的深层机制还不清楚. 目的:利用 transwel 技术,在体外建立一种成骨与破骨细胞的新型共育体系,为深入研究骨重建中成骨与破骨细胞的相互作用提供成熟的实验模型. 方法:采用 MC3T3-E1成骨样细胞株与RAW264.7破骨前体细胞株,进行体外成骨与破骨细胞的诱导分化,并利用Transwel 共培养板(0.4μm聚酯膜)建立成骨与破骨细胞的共育体系.共培养6 d后,通过测定细胞活性和碱性磷酸酶(ALP)活力分析成骨细胞的增殖和分化活性,利用抗酒石酸酸性磷酸酶(TRAP)染色、甲苯胺蓝(TB)染色、TRAP活性测定及扫描电镜技术观察破骨细胞的分化及骨吸收功能. 结果与结论:共培养体系中成骨样细胞的无限增殖能力减弱,而分化活性明显增强,同时破骨前体细胞被诱导分化为成熟的破骨细胞,并具有一定的骨吸收功能.因此,该共培养体系可用于骨重建中成骨与破骨细胞间信号通路的深层研究.
배경:음소주지,골중건시골조직중중요적생물학반응과정,기중성골세포여파골세포발휘료관건작용.단목전,관우골중건중성골여파골세포간신호전체적심층궤제환불청초. 목적:이용 transwel 기술,재체외건립일충성골여파골세포적신형공육체계,위심입연구골중건중성골여파골세포적상호작용제공성숙적실험모형. 방법:채용 MC3T3-E1성골양세포주여RAW264.7파골전체세포주,진행체외성골여파골세포적유도분화,병이용Transwel 공배양판(0.4μm취지막)건립성골여파골세포적공육체계.공배양6 d후,통과측정세포활성화감성린산매(ALP)활력분석성골세포적증식화분화활성,이용항주석산산성린산매(TRAP)염색、갑분알람(TB)염색、TRAP활성측정급소묘전경기술관찰파골세포적분화급골흡수공능. 결과여결론:공배양체계중성골양세포적무한증식능력감약,이분화활성명현증강,동시파골전체세포피유도분화위성숙적파골세포,병구유일정적골흡수공능.인차,해공배양체계가용우골중건중성골여파골세포간신호통로적심층연구.
@@@@BACKGROUND:It is wel known that bone remodeling is an important biological process in which osteoblasts and osteoclasts play the critical role. However, the detailed mechanisms of osteoblast-osteoclast communication in bone remodeling remain unclear. @@@@OBJECTIVE:To in vitro establish a novel co-culture system for interaction of mouse osteoblasts and osteoclasts using the Transwel technology. @@@@METHODS:Osteoblastic cel line MC3T3-E1 and preosteoclasts RAW264.7 were selected, and then the cel s were induced to differentiate into osteoblasts and osteoclasts, respectively. The co-culture system for osteoblasts and osteoclasts interaction was established with Transwel co-culture plate (0.4 μm polyester film), and the osteoblasts and osteoclasts were co-cultured for 6 days. After that, the proliferation and differentiation activities of osteoblasts were analyzed through testing the cel activity and alkaline phosphatase activity, and the differentiation and bone resorption of osteoclasts were observed using tartrate-resistant acid phosphatase staining, toluidine blue staining, tartrate-resistant acid phosphatase activity detecting and scanning microscope technology. @@@@RESULTS AND CONCLUSION:In the co-culture system, the unlimited proliferation ability of osteoblasts was decreased, while the differentiation activity was increased. Meanwhile, the preosteoclasts could differentiate into mature osteoclasts with bone resorption function. Thus, this co-culture system can be applied in the further study of osteoblast-osteoclast communication in bone remodeling.