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[双辉等离子体渗氮技术改性的一种近β型钛合金表面的初始成骨细胞功能]

[Initial osteoblast functions on a type of near β-type titanium alloys surfaces modified by the double glow plasma nitriding technology].

作者信息

Qu Y H, Li F L, Wen K, Wang W

机构信息

Department of Prosthodontics, Affiliated People's Hospital, Shanxi Medical University, Taiyuan 030012, China (Present address: Department of Oral Medicine, Shanxi Medical University, Taiyuan 030000, China).

Department of Prosthodontics, Affiliated People's Hospital, Shanxi Medical University, Taiyuan 030012, China.

出版信息

Zhonghua Kou Qiang Yi Xue Za Zhi. 2017 Feb 9;52(2):132-136. doi: 10.3760/cma.j.issn.1002-0098.2017.02.016.

Abstract

To evaluate the adhesion, proliferation, alkaline phosphatase (ALP) activity and the expression of osteogenesis-related genes and osteoprotegerin (OPG)/receptor activator of NF-κB ligand (RANKL) of osteoblast-like cells on a type of near β-type titanium alloys (Ti-5Zr-3Sn-5Mo-15Nb, TLM) surfaces modified by the double glow plasma nitriding technology, and to investigate the effect of the modified surfaces on the initial functions of osteoblast-like cells. The surfaces of TLM were modified by the double glow plasma nitriding technology. TLM surfaces without modification were used as control. Cell morphology was observed with scanning electron microscopy (SEM). Methyl thiazolyl tetrazolium (MTT) method was used to measure cell proliferation. Cell ALP activity was evaluated by using reagent kits. The mRNA expression of Runt-related transcription factor-2 (RUNX2), typeⅠcollagen alpha 1 chain (COLⅠ α1) and OPG/RANKL were examined by quantitative real-time PCR(qRT-PCR). Four hour following cell alture, cells on modified surfaces extend filopodia and intercellular junction was tight. Three days later, cell proliferation (0.277±0.007) was significantly higher than that in control group (0.249±0.004) (0.01). After two weeks, ALP activity on TLM modified layer (173.6±1.89) was significantly higher than that on unmodified TLM (162.6±2.4) (0.01). The mRNA expression of osteoblast marker RUNX2, COLⅠα1 were stronger than that in control group (0.05). The expression of OPG mRNA was higher than that in control group (0.01), and RANKL mRNA expression was significantly lower than that in control group (0.05). The TLM surface modified by the double glow plasma nitriding technology has a positive effect on osteoblasts initial adhesion, proliferation and differentiation, and it can also improve expression of OPG mRNA and has an inhibitory effect on RANKL mRNA expression of osteoblasts.

摘要

为评价经双层辉光等离子体渗氮技术改性的一种近β型钛合金(Ti-5Zr-3Sn-5Mo-15Nb,TLM)表面上类成骨细胞的黏附、增殖、碱性磷酸酶(ALP)活性以及成骨相关基因和骨保护素(OPG)/核因子κB受体活化因子配体(RANKL)的表达,并研究改性表面对类成骨细胞初始功能的影响。采用双层辉光等离子体渗氮技术对TLM表面进行改性,未改性的TLM表面作为对照。用扫描电子显微镜(SEM)观察细胞形态。采用甲基噻唑基四氮唑(MTT)法检测细胞增殖。使用试剂盒评估细胞ALP活性。通过定量实时聚合酶链反应(qRT-PCR)检测Runt相关转录因子2(RUNX2)、Ⅰ型胶原α1链(COLⅠα1)和OPG/RANKL的mRNA表达。细胞接种4小时后,改性表面的细胞伸出丝状伪足,细胞间连接紧密。3天后,细胞增殖率(0.277±0.007)显著高于对照组(0.249±0.004)(P<0.01)。2周后,TLM改性层上的ALP活性(173.6±1.89)显著高于未改性的TLM(162.6±2.4)(P<0.01)。成骨细胞标志物RUNX2、COLⅠα1的mRNA表达强于对照组(P<0.05)。OPG mRNA的表达高于对照组(P<0.01),而RANKL mRNA表达显著低于对照组(P<0.05)。经双层辉光等离子体渗氮技术改性的TLM表面对成骨细胞的初始黏附、增殖和分化具有积极作用,还可提高OPG mRNA的表达,并对成骨细胞RANKL mRNA表达具有抑制作用。

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