CAJ | 학술논문

目的探讨在体外条件下低频电磁场诱导大鼠骨髓间充质干细胞向软骨细胞分化的可行性.方法大鼠骨髓间充质于细胞传代至第5代后,采用微团培养法在含成纤维生长因子-2和转化生长因子-β3的培养基中生长,并给予正弦波电磁场(1.0 mT,50 Hz)干预.3周后进行阿利新蓝染色以检测软骨基质生成量,采用荧光定量聚合酶链反应(FQ-PCR)技术检测软骨特异性蛋白的基因表达水平,并使用二甲基-亚甲蓝(DMMB)染料结合法评估细胞微团中糖胺多糖水平.结果在电磁场和生长因子的协同作用下,大鼠骨髓间充质干细胞(BMSCs)细胞微团向软骨分化,细胞微团中Ⅱ型、X型胶原及蛋白多糖基因表达水平显著增加,而性别决定区Y框蛋白9(SOX9)的表达没有明显变化.与非暴磁组比较,暴磁组细胞糖胺多糖(GAG)/DNA比例较高(3.108±0.341).结论电磁场促进大鼠BMSCs成软骨分化可能与细胞表达Ⅱ、X型胶原及糖胺多糖增多有关.电磁场在生长因子的协同作用下可诱导及维持间充质干细胞成软骨分化,但单因素电磁场刺激不能诱导大鼠骨髓间充质干细胞成软骨分化.
목적 탐토재체외조건하저빈전자장유도대서골수간충질간세포향연골세포분화적가행성.방법 대서골수간충질우세포전대지제5대후,채용미단배양법재함성섬유생장인자-2화전화생장인자-β3적배양기중생장,병급여정현파전자장(1.0 mT,50 Hz)간예.3주후진행아리신람염색이검측연골기질생성량,채용형광정량취합매련반응(FQ-PCR)기술검측연골특이성단백적기인표체수평,병사용이갑기-아갑람(DMMB)염료결합법평고세포미단중당알다당수평.결과 재전자장화생장인자적협동작용하,대서골수간충질간세포(BMSCs)세포미단향연골분화,세포미단중Ⅱ형、X형효원급단백다당기인표체수평현저증가,이성별결정구Y광단백9(SOX9)적표체몰유명현변화.여비폭자조비교,폭자조세포당알다당(GAG)/DNA비례교고(3.108±0.341).결론 전자장촉진대서BMSCs성연골분화가능여세포표체Ⅱ、X형효원급당알다당증다유관.전자장재생장인자적협동작용하가유도급유지간충질간세포성연골분화,단단인소전자장자격불능유도대서골수간충질간세포성연골분화.
Objective To determine the chondrogenic potential of mesenchymal stem cells (BMSCs) under exposure of extremely low frequency electromagnetic fields (ELF-EMF) and discuss the application prospect of EMF in cartilage tissue engineering.Methods Cell pellets of rat bone marrow-derived BMSCs were cultured in vitro under the addition of fibroblast growth factor 2 (FGF-2) and transforming growth factor-β3 (TGF-β3).Pellet cultures were exposed to sinusoidal ELF-EMF (1.0 mT,50 Hz).After 3 weeks of inductive differentiation culture,chondrogenesis was detected by alcian blue staining,and quantitative real-time polymerase chain reaction (real-time PCR) was used for detection of chondrogenesis related proteins.Glycosaminoglycan (GAG) contents of pellet cultures were determined using the dimethylmethylene blue (DMMB) dye-binding assay.Results With the effects of growth factors,EMF could significantly promote chondrogenic differentiation of BMSCs pellet cultures,and the gene expression of collagen type Ⅱ,X and aggrecan in rat BMSCs was also significantly up-regulated.The gene expression of sex determining region Y box 2 (SOX9) did not show significant difference between EMF exposed groups and nonexposed groups.The glycosaminoglycan (GAG)/DNA ratio of EMF exposed pellet cultures reached 3.108 ±0.341,which was significantly higer than those unexposed pellet cultures.Conclusion Increased expression of collagen type Ⅱ,X and GAG content in pellet cultures under EMF exposure may contribute to the chonderogenic differentiation of rat BMSCs.EMF was able to stimulate and maintain chondrogenic differentiation of rat BMSCs under influence of growth factors.EMF alone,however,could not induce chondrogenic differentiation.