Liang N W, Shi L, Huang Y, Deng X L
Department of Prosthodontics, Peking University School and Hospital of Stomatology & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China.
Department of Geriatric Dentistry, Peking University School and Hospital of Stomatology & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2017 Feb 18;49(1):43-8.
To study the role of different scale structure of Ti implants on the biological behaviors of human umbilical vein endothelial cell (HUVECs) and to reveal the role of material surface topographical features on peri-implant angiogenesis.
Titanium (Ti) discs with different surface structures (Ti discs with smooth surface, Ti discs with nano scale structure, Ti discs with micro scale structure and Ti discs with micro/nano scale structure, named as SM-Ti, Nano-Ti, Micro-Ti and Micro/Nano-Ti, respectively) were prepared and their surface topographical features were confirmed via scanning electron microscopy (SEM) observation. HUVECs were cultured on these Ti discs. Biological outcomes of HUVECs on different surfaces were carried out, including cell adhesive capacity, proliferation, vascular endothelial growth factor (VEGF) production and intracellular expression of Ca(2+).
The results of SEM images and immunofluorescence double staining of rhodamine-phalloidin and DAPI showed that compared with the SM-Ti and Nano-Ti group, the adhesive capacity and proliferation behavior of HUVECs on the surfaces of Micro-Ti and Micro/Nano-Ti was decreased. The results of culturing HUVECs on different groups of Ti discs after 24 hours showed that the cells number grew from (18±4) to (42±6)/ vision on SM-Ti, (28±6) to (52±10)/vision on Nano-Ti, (20±4) to (21±6)/vision on Micro-Ti and (16±4) to (18±6)/vision on Micro/Nano-Ti. Moreover, compared with the adhesion and proliferation of HUVECs on SM-Ti group and Nano-Ti, the adhesion and proliferation of HUVECs on Micro-Ti group and Micro/Nano-Ti group was significantly reduced (P<0.05).The results of enzyme-linked immunosorbent assay (ELISA) showed that the VEGF productions of SM-Ti, Nano-Ti, Micro-Ti and Micro/Nano-Ti were (690±35) ng/L, (560±20) ng/L, (474±43) ng/L and (517±29) ng/L, respectively. Moreover, compared with the VEGF production of HUVECs on SM-Ti group, the VEGF production of HUVECs on Micro-Ti group and Micro/Nano-Ti group was significantly reduced (P<0.05). The results of Ca(2+) ion detection showed that the Ca(2+) expression of HUVECs on Micro-Ti and Micro/Nano-Ti was significantly higher than that on the surface of SM-Ti and Nano-Ti. These results implied that the over expressed Ca(2+) might contributed to the impaired biological function of HUVECs on Micro-Ti and Micro/Nano-Ti.
Different topographical features on titanium influenced the biological behaviors of the HUVECs, which may illustrate how topographical features of Ti implant affect peri-implant angiogenesis. These results also suggest that the biological behaviors of HUVECs might be relative to the changed expression of intracellular Ca(2+).
研究钛种植体不同尺度结构对人脐静脉内皮细胞(HUVECs)生物学行为的作用,并揭示材料表面形貌特征对种植体周围血管生成的作用。
制备具有不同表面结构的钛(Ti)盘(表面光滑的Ti盘、具有纳米尺度结构的Ti盘、具有微米尺度结构的Ti盘和具有微/纳米尺度结构的Ti盘,分别命名为SM-Ti、Nano-Ti、Micro-Ti和Micro/Nano-Ti),并通过扫描电子显微镜(SEM)观察确认其表面形貌特征。将HUVECs培养在这些Ti盘上。对不同表面上的HUVECs进行生物学检测,包括细胞黏附能力、增殖、血管内皮生长因子(VEGF)产生以及细胞内Ca(2+)的表达。
SEM图像以及罗丹明 - 鬼笔环肽和DAPI免疫荧光双染结果显示,与SM-Ti和Nano-Ti组相比,HUVECs在Micro-Ti和Micro/Nano-Ti表面的黏附能力和增殖行为降低。在不同组Ti盘上培养HUVECs 24小时的结果显示,细胞数量在SM-Ti上从(18±4)增长至(42±6)/视野,在Nano-Ti上从(28±6)增长至(52±10)/视野,在Micro-Ti上从(20±4)增长至(21±6)/视野,在Micro/Nano-Ti上从(16±4)增长至(18±6)/视野。此外,与HUVECs在SM-Ti组和Nano-Ti上的黏附及增殖相比,HUVECs在Micro-Ti组和Micro/Nano-Ti组上的黏附及增殖显著降低(P<0.05)。酶联免疫吸附测定(ELISA)结果显示,SM-Ti、Nano-Ti、Micro-Ti和Micro/Nano-Ti的VEGF产生量分别为(690±35) ng/L、(560±20) ng/L、(474±43) ng/L和(517±29) ng/L。此外,与HUVECs在SM-Ti组的VEGF产生量相比,HUVECs在Micro-Ti组和Micro/Nano-Ti组的VEGF产生量显著降低(P<0.05)。Ca(2+)离子检测结果显示,HUVECs在Micro-Ti和Micro/Nano-Ti上的Ca(2+)表达显著高于在SM-Ti和Nano-Ti表面。这些结果表明,过量表达的Ca(2+)可能导致HUVECs在Micro-Ti和Micro/Nano-Ti上的生物学功能受损。
钛表面不同的形貌特征影响了HUVECs的生物学行为,这可能解释了Ti种植体的形貌特征如何影响种植体周围血管生成。这些结果还表明,HUVECs的生物学行为可能与细胞内Ca(2+)表达的变化有关。