Orthopaedic Department, The Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China.
Mol Med Rep. 2017 Nov;16(5):7591-7596. doi: 10.3892/mmr.2017.7512. Epub 2017 Sep 19.
The aim of the present study was to investigate the effects of cobalt nanoparticles (CoNPs) on the proliferation and differentiation of human osteoblasts in vitro, and to investigate the molecular mechanisms via which CoNPs affect proliferation and differentiation of osteoblasts. The MG‑63 human osteoblast cell line was treated with different concentrations of CoNPs for 12 to 48 h in vitro. At each time point, cell morphology was observed and an MTT assay was performed to assess cell viability. Alkaline phosphatase (ALP), osteocalcin (BGLAP), collagen I (COL I), osteoprotegerin (OPG) and receptor activator of nuclear factor κ‑B ligand mRNA expression levels, and ALP, BGLAP and COL protein expression levels, were assessed by reverse transcription‑quantitative polymerase chain reaction and western blotting, respectively. The viability of MG‑63 cells decreased significantly after treatment with CoNPs. As CoNP concentration increased, a higher growth inhibition and cell death was observed. Compared with CoNPs, treatment with the same concentration of Co2+ may have a greater inhibitory effect on the growth of MG‑63 cells. CoNPs affected the mRNAs expression levels of ALP, BGLAP, COL I and OPG in MG‑63 cells, and reduced the protein expression levels of ALP, BGLAP and COL I. In conclusion, the present study demonstrated that CoNPs induce cytotoxic effects on MG‑63 cells by markedly reducing cell viability and inducing cell death at high concentrations. In addition, CoNPs may inhibit the function and differentiation of osteoblasts by affecting the mRNA and protein expression levels of associated genes. The results of the present study indicate that CoNPs may serve an important role in the aseptic loosening mechanism following total joint replacement surgery, particularly in situations where metal on metal prostheses are used. Further study into inhibiting this effect is required.
本研究旨在探讨钴纳米粒子（CoNPs）对人成骨细胞体外增殖和分化的影响，并探讨 CoNPs 影响成骨细胞增殖和分化的分子机制。将 MG-63 人成骨细胞系用不同浓度的 CoNPs 处理 12 至 48 小时。在每个时间点，观察细胞形态并用 MTT 法评估细胞活力。通过逆转录-定量聚合酶链反应和 Western blot 分别评估碱性磷酸酶（ALP）、骨钙素（BGLAP）、胶原 I（COL I）、骨保护素（OPG）和核因子 κB 配体受体激活剂（RANKL）mRNA 表达水平以及 ALP、BGLAP 和 COL 蛋白表达水平。用 CoNPs 处理后，MG-63 细胞的活力明显下降。随着 CoNP 浓度的增加，观察到更高的生长抑制和细胞死亡。与 CoNPs 相比，用相同浓度的 Co2+处理可能对 MG-63 细胞的生长有更大的抑制作用。CoNPs 影响 MG-63 细胞中 ALP、BGLAP、COL I 和 OPG 的 mRNAs 表达水平，并降低 ALP、BGLAP 和 COL I 的蛋白表达水平。综上所述，本研究表明 CoNPs 通过显著降低细胞活力和在高浓度下诱导细胞死亡，对 MG-63 细胞产生细胞毒性作用。此外，CoNPs 可能通过影响相关基因的 mRNA 和蛋白表达水平来抑制成骨细胞的功能和分化。本研究结果表明，CoNPs 在全关节置换手术后的无菌性松动机制中可能发挥重要作用，特别是在使用金属对金属假体的情况下。需要进一步研究抑制这种作用。
