PDF(Sci-hub)
Abstract：
OBJECTIVE: Zinc (Zn), an essential trace element in the body, has stable chemical properties, excellent osteogenic ability and moderate immunomodulatory property. In the present study, a Zn-incorporated TiO2 nanotube (TNT) was fabricated on titanium (Ti) implant material. We aimed to evaluate the influence of nano-scale topography and Zn on behaviors of murine RAW 264.7 macrophages. Moreover, the effects of Zn-incorporated TNT surface-regulated macrophages on the behaviors and osteogenic differentiation of murine MC3T3-E1 osteoblasts were also investigated.
METHODS: TNT coatings were firstly fabricated on a pure Ti surface using anodic oxidation, and then nano-scale Zn particles were incorporated onto TNTs by the hydrothermal method. Surface topography, chemical composition, roughness, hydrophilicity, Zn release pattern and protein adsorption ability of the Zn-incorporated TiO2 nanotube surface were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS), surface profiler, contact angle test, Zn release test and protein adsorption test. The cell behaviors and both pro-inflammatory (M1) and pro-regenerative (M2) marker gene and protein levels in macrophages cultured on Zn-incorporated TNTs surfaces with different TNT diameters were detected. The supernatants of macrophages were extracted and preserved as conditioned medium (CM). Furthermore, the behaviors and osteogenic properties of osteoblasts cultured in CM on various surfaces were evaluated.
RESULTS: The release profile of Zn on Zn-incorporated TNT surfaces revealed a controlled release pattern. Macrophages cultured on Zn-incorporated TNT surfaces displayed enhanced gene and protein expression of M2 markers, and M1 markers were moderately inhibited, compared with the LPS group (the inflammation model). When cultured in CM, osteoblasts cultured on Zn-incorporated TNTs showed strengthened cell proliferation, adhesion, osteogenesis-related gene expression, alkaline phosphatase activity and extracellular mineralization, compared with their TNT counterparts and the Ti group.
CONCLUSIONS: This study suggests that the application of Zn-incorporated TNT surfaces may establish an osteogenic microenvironment and accelerate bone formation. It provided a promising strategy of Ti surface modification for a better applicable prospect.
METHODS: TNT coatings were firstly fabricated on a pure Ti surface using anodic oxidation, and then nano-scale Zn particles were incorporated onto TNTs by the hydrothermal method. Surface topography, chemical composition, roughness, hydrophilicity, Zn release pattern and protein adsorption ability of the Zn-incorporated TiO2 nanotube surface were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS), surface profiler, contact angle test, Zn release test and protein adsorption test. The cell behaviors and both pro-inflammatory (M1) and pro-regenerative (M2) marker gene and protein levels in macrophages cultured on Zn-incorporated TNTs surfaces with different TNT diameters were detected. The supernatants of macrophages were extracted and preserved as conditioned medium (CM). Furthermore, the behaviors and osteogenic properties of osteoblasts cultured in CM on various surfaces were evaluated.
RESULTS: The release profile of Zn on Zn-incorporated TNT surfaces revealed a controlled release pattern. Macrophages cultured on Zn-incorporated TNT surfaces displayed enhanced gene and protein expression of M2 markers, and M1 markers were moderately inhibited, compared with the LPS group (the inflammation model). When cultured in CM, osteoblasts cultured on Zn-incorporated TNTs showed strengthened cell proliferation, adhesion, osteogenesis-related gene expression, alkaline phosphatase activity and extracellular mineralization, compared with their TNT counterparts and the Ti group.
CONCLUSIONS: This study suggests that the application of Zn-incorporated TNT surfaces may establish an osteogenic microenvironment and accelerate bone formation. It provided a promising strategy of Ti surface modification for a better applicable prospect.
摘要：
目标：锌（Zn），体内必需的微量元素,具有稳定的化学性质，良好的成骨能力和中等的免疫调节性能。在本研究中,在钛(Ti)植入材料上制造掺入Zn的TiO2纳米管(TNT)。我们旨在评估纳米尺度形貌和锌对小鼠RAW264.7巨噬细胞行为的影响。此外，还研究了掺入Zn的TNT表面调节巨噬细胞对小鼠MC3T3-E1成骨细胞的行为和成骨分化的影响。
方法：首先使用阳极氧化在纯Ti表面上制备TNT涂层，然后通过水热法将纳米级Zn颗粒掺入到TNTs上。表面形貌,化学成分,粗糙度,亲水性,通过扫描电子显微镜(SEM)表征了掺入Zn的TiO2纳米管表面的Zn释放模式和蛋白质吸附能力,X射线衍射(XRD)和X射线光电子能谱(XPS),表面轮廓仪,接触角试验,锌释放试验和卵白质吸附试验。检测了在不同直径的Zn掺入的TNT表面上培养的巨噬细胞中的细胞行为以及促炎（M1）和促再生（M2）标记基因和蛋白质水平。提取巨噬细胞的上清液并保存为条件培养基(CM)。此外，评价了在不同表面的CM中培养的成骨细胞的行为和成骨特性。
结果：Zn在掺入Zn的TNT表面上的释放曲线显示出受控的释放模式。在掺入Zn的TNT表面上培养的巨噬细胞显示出M2标记的基因和蛋白质表达增强，M1标记受到中度抑制，与LPS组(炎症模型)比较。当在CM中培养时，在掺入Zn的TNTs上培养的成骨细胞显示出增强的细胞增殖，附着力,成骨相关基因表达，碱性磷酸酶活性和细胞外矿化，与他们的TNT同行和Ti组相比。
结论：这项研究表明，掺入Zn的TNT表面的应用可能会建立成骨微环境并加速骨形成。为钛表面改性提供了一种有前景的策略，具有更好的应用前景。
方法：首先使用阳极氧化在纯Ti表面上制备TNT涂层，然后通过水热法将纳米级Zn颗粒掺入到TNTs上。表面形貌,化学成分,粗糙度,亲水性,通过扫描电子显微镜(SEM)表征了掺入Zn的TiO2纳米管表面的Zn释放模式和蛋白质吸附能力,X射线衍射(XRD)和X射线光电子能谱(XPS),表面轮廓仪,接触角试验,锌释放试验和卵白质吸附试验。检测了在不同直径的Zn掺入的TNT表面上培养的巨噬细胞中的细胞行为以及促炎（M1）和促再生（M2）标记基因和蛋白质水平。提取巨噬细胞的上清液并保存为条件培养基(CM)。此外，评价了在不同表面的CM中培养的成骨细胞的行为和成骨特性。
结果：Zn在掺入Zn的TNT表面上的释放曲线显示出受控的释放模式。在掺入Zn的TNT表面上培养的巨噬细胞显示出M2标记的基因和蛋白质表达增强，M1标记受到中度抑制，与LPS组(炎症模型)比较。当在CM中培养时，在掺入Zn的TNTs上培养的成骨细胞显示出增强的细胞增殖，附着力,成骨相关基因表达，碱性磷酸酶活性和细胞外矿化，与他们的TNT同行和Ti组相比。
结论：这项研究表明，掺入Zn的TNT表面的应用可能会建立成骨微环境并加速骨形成。为钛表面改性提供了一种有前景的策略，具有更好的应用前景。
