论文部分内容阅读
目的观察脉冲电磁场(pulsed electromagnetic fields,PEMF)对脱钙骨基质(demineralized bone matrix,DBM)诱导人骨髓间充质干细胞(human mesenchymal stem cells,hMSCs)成骨分化的影响,探讨可能的发生机制。方法提取1例健康14周岁男性骨髓间充质干细胞,培养至第3代,以1×104/孔接种至8块24孔板,以1×105/孔接种至2块6孔板,各分成4组:细胞对照组(C组)、细胞+材料组(CD组)、细胞+脉冲电磁场组(CP组)和细胞+材料+脉冲电磁场组(CDP组)。CD组和CDP组加入一块5mm×5mm×3mm大小的DBM,CP组和CDP组给予PEMF(频率15Hz,场强5Gs)照射;各组分别于种植后第1、7、14、21天进行碱性磷酸酶(ALP)活性、骨钙素(OC)浓度等指标检测,在第21天进行钙结节茜素红染色,光镜观察。结果CD组、CP组和CDP组ALP活性及OC浓度在种植7d后均明显升高(P<0.01);在14d时CD组和CDP组ALP活性达到最高值,第7、14天时CDP组ALP活性较CD组、CP组均显著升高(P<0.01),仅在第14天时CD组ALP活性较CP组显著升高(P<0.01);CDP组OC值在7、14、21d时均较CD组、CP组显著升高(P<0.01),仅在第21天时CD组OC浓度较CP组显著升高(P<0.01);21d时形态学观察显示钙结节数量CDP组明显多于CD组和CP组(P<0.01),CD组明显多于CP组(P<0.01)。结论PEMF与DBM联合对hMSCs的成骨诱导要强于PEMF或DBM的单独作用,且随着磁场应用时间的延长更加明显。另外PEMF对DBM诱导hMSCs成骨分化具有明显的协同效应,可能是通过PEMF增强hMSCs对BMPs等成骨活性因子的反应性来实现的。
Objective To investigate the effect of pulsed electromagnetic fields (PEMF) on the osteogenic differentiation of human mesenchymal stem cells (hMSCs) induced by demineralized bone matrix (DBM), and to explore the possible mechanism. METHODS: One male BMSC, aged 14 years, was isolated and cultured to the third passage. The cells were inoculated into 8 24-well plates at 1 × 104 / well and inoculated into 2 6-well plates at 1 × 105 / well, Four groups: control group (C group), cells + material group (CD group), cells + pulsed electromagnetic field group (CP group) and cells + material + pulsed electromagnetic field group (CDP group). CDM group and CDP group were added to a piece of DBM 5mm × 5mm × 3mm size, CP group and CDP group were given PEMF (frequency 15Hz, field intensity 5Gs) irradiation; each group were planted on the 1st, 14th, (ALP) activity and osteocalcin (OC) were measured. On day 21, alizarin red staining of calcium nodules was observed by light microscopy. Results The ALP activity and OC concentration in CD group, CP group and CDP group were significantly increased after 7 days of planting (P <0.01). The ALP activity of CD group and CDP group reached the highest value on the 14th day. (P <0.01). The ALP activity in CD group was significantly higher than that in CP group only on the 14th day (P <0.01), while the OC value in CDP group was on the 7th, 14th and 21st day Compared with CD group, CP group was significantly increased (P <0.01), CD concentration in CD group was significantly higher than that in CP group only on the 21st day (P <0.01), and on the 21st day, the number of calcium nodules in the CDP group was significantly higher In CD group and CP group (P <0.01), CD group was more than that in CP group (P <0.01). Conclusion The osteogenic induction of hMSCs by PEMF combined with DBM is stronger than that of PEMF or DBM alone, and it is more obvious with the prolongation of magnetic field. In addition, PEMF has obvious synergistic effect on DBM-induced osteogenic differentiation of hMSCs, which may be achieved by PEMF enhancing the reactivity of hMSCs to osteogenic active factors such as BMPs.