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人工关节置换应用中类金刚石碳(DLC)磨损颗粒的尺寸依赖性细胞毒性评估。

Evaluation of the Size-Dependent Cytotoxicity of DLC (Diamondlike Carbon) Wear Debris in Arthroplasty Applications.

作者信息

Liao T T, Deng Q Y, Li S S, Li X, Ji L, Wang Q, Leng Y X, Huang N

机构信息

Key Laboratory for Advanced Technologies of Materials, Ministry of Education, School of Materials Science and Engineering, Southwest Jiaotong University, Chengdu, Sichuan 610031, China.

出版信息

ACS Biomater Sci Eng. 2017 Apr 10;3(4):530-539. doi: 10.1021/acsbiomaterials.6b00618. Epub 2017 Mar 27.

Abstract

Patients with DLC (diamond like carbon)-coated artificial joints may be exposed to a wide size range of DLC wear debris (DW). In this study, the cytotoxicity of DW of different size ranges (0-0.22, 0.22-0.65, 0.65-1.0, and 1.0-5.0 μm) was evaluated. The microstructure and physical characteristics of DW were investigated by Raman spectroscopy, transmission electron microscopy (TEM), scanning electron microscope (SEM), and dynamic light scattering (DLS). Macrophages, osteoblasts, and fibroblasts were incubated with DW of different size ranges respectively followed by cytotoxicity evaluations of inflammatory cytokines, alkaline phosphatase (ALP) assays, and related signal protein expression analysis. The results showed that, except for the size range of 0-0.22 μm, DW cytotoxicity showed a size-dependent (0.22-5.0 μm) decrease with increasing size. Within the range of 0.22-5.0 μm, DW of larger size resulted in lessened inflammatory response and enhanced osteoblastogenesis and fibrogenesis, with increased viability of cells (macrophages, osteoblasts, and fibroblasts), better morphology, less release of pro-inflammatory factors and more release of anti-inflammatory factors. The results demonstrated that DW sizes below 0.22 μm had less negative effects on cell adhesion and growth because of the BSA (bovine serum albumin) encapsulation effect. These findings provide valuable knowledge about the comprehensive mechanism of promotion of inflammatory response and inhibition of osteoblastogenesis and fibrogenesis induced by DW. In conclusion, an effective system of biocompatibility evaluation for different sizes of DW was derived.

摘要

患有类金刚石碳(DLC)涂层人工关节的患者可能会接触到各种尺寸范围的DLC磨损颗粒(DW)。在本研究中,评估了不同尺寸范围(0 - 0.22、0.22 - 0.65、0.65 - 1.0和1.0 - 5.0μm)的DW的细胞毒性。通过拉曼光谱、透射电子显微镜(TEM)、扫描电子显微镜(SEM)和动态光散射(DLS)研究了DW的微观结构和物理特性。分别用不同尺寸范围的DW孵育巨噬细胞、成骨细胞和成纤维细胞,然后进行炎性细胞因子的细胞毒性评估、碱性磷酸酶(ALP)测定和相关信号蛋白表达分析。结果表明,除了0 - 0.22μm尺寸范围外,DW细胞毒性呈现出尺寸依赖性(0.22 - 5.0μm),随着尺寸增加而降低。在0.22 - 5.0μm范围内,较大尺寸的DW导致炎症反应减轻,成骨和纤维化增强,细胞(巨噬细胞、成骨细胞和成纤维细胞)活力增加,形态更好,促炎因子释放减少,抗炎因子释放增加。结果表明,由于牛血清白蛋白(BSA)包封作用,尺寸低于0.22μm的DW对细胞黏附和生长的负面影响较小。这些发现为DW诱导的炎症反应促进以及成骨和纤维化抑制的综合机制提供了有价值的知识。总之,得出了一个针对不同尺寸DW的有效生物相容性评估系统。

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