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首页> 外文期刊>Biomolecular engineering >Influence of manganese ions on cellular behavior of human osteoblasts in vitro.
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Influence of manganese ions on cellular behavior of human osteoblasts in vitro.

机译：锰离子对人成骨细胞体外细胞行为的影响。

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Divalent cations like Mn(2+) are known to strongly influence the integrin affinity to ligands and - in consequence - cell adhesion to extracellular matrix proteins. Therefore, divalent cation supplementation of biomaterials could be a promising approach to improve the ingrowth and the integration of implants. We were interested, whether manganese ions affect cellular functions like spreading, proliferation as well as gene expression in human osteoblasts. MG-63 osteoblastic cells were cultured in DMEM with 10% FCS. MnCl(2) was added at a concentration range of 0.01-0.5mM for 24h and 48h. Spreading (cell area in mum(2)) of PKH26-stained cells (cell membrane dye) was analyzed using confocal microscopy. Cell proliferation was measured by flow cytometry. Quantification of the phosphorylation status of signaling proteins was estimated using the Bio-Plex 200 system. Gene expression of osteogenic markers at the mRNA and protein level was analyzed by quantitative real time RT-PCR and Western blot, respectively.The results demonstrated that at higher concentrations of Mn(2+) cells revealed a spindle shaped morphology. Further analyses indicated a reduced spreading, proliferation as well as phosphorylation of signaling proteins due to the influence of Mn(2+) in a concentration-dependent manner. Although expression of bone sialo protein (BSP) at the mRNA level increased both after 24h and 48h in the presence of manganese, no increased expression of BSP was detected at the protein level. The expression of alkaline phosphatase (ALP) and collagen 1 (Col 1) mRNA decreased at >0.1mM MnCl(2). We speculate that the effect of manganese cations on cell functions is strongly concentration-dependent and the release of manganese when incorporated in a biomaterial surface has to be thoroughly adjusted.
机译：已知像Mn（2+）这样的二价阳离子会强烈影响整联蛋白对配体的亲和力，进而影响细胞对细胞外基质蛋白的粘附。因此，生物材料的二价阳离子补充可能是改善植入物向内生长和整合的有前途的方法。我们感兴趣的是，锰离子是否会影响人类成骨细胞中细胞的功能，例如扩散，增殖以及基因表达。 MG-63成骨细胞在含10％FCS的DMEM中培养。 MnCl（2）以0.01-0.5mM的浓度范围添加24h和48h。使用共聚焦显微镜分析了PKH26染色的细胞（细胞膜染料）的扩散情况（在mum（2）中的细胞面积）。细胞增殖通过流式细胞术测量。使用Bio-Plex 200系统评估信号蛋白磷酸化状态的定量。通过定量实时RT-PCR和Western blot分别分析了成骨标记在mRNA和蛋白质水平的基因表达。结果表明，在较高浓度的Mn（2+）细胞中显示出纺锤形的形态。进一步的分析表明，由于Mn（2+）的浓度依赖性，减少了信号蛋白的扩散，增殖以及磷酸化。尽管在锰存在下24h和48h后，骨唾液蛋白（BSP）在mRNA水平上的表达均增加，但在该蛋白水平上未检测到BSP表达的增加。碱性磷酸酶（ALP）和胶原蛋白1（Col 1）mRNA的表达在> 0.1mM MnCl（2）时降低。我们推测，锰阳离子对细胞功能的影响强烈地依赖于浓度，当掺入生物材料表面时锰的释放必须进行彻底调整。

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