Sugiatno Erwan, Samsudin Abdul R, Sosroseno Wihaskoro
Department of Prosthodontics, Faculty of Dentistry, Gadjah Mada University, Yogyakarta - Indonesia and School of Dental Sciences, University Sains Malaysia, Kota Bharu - Malaysia.
J Appl Biomater Biomech. 2009 Jan-Apr;7(1):29-33.
The aim of this study was to test the hypothesis that the proliferation of hydroxyapatite (HA)-induced human osteoblast cell line (HOS cells) may be up-regulated by exogenous nitric oxide (NO).
HOS cells were cultured on the surface of HA with or without the presence of a NO donor, S-nitroso acetyl penicillamine (SNAP) or nitroso acetyl penicillamine (NAP). 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) known as carboxy PTIO, a NO scavenger, was added in the cell cultures with or without the presence of SNAP. The cells were also pre-treated with L-N5-(1-iminoethyl)orthinine hydrochloride (L-NIO), an endothelial nitric oxide synthase (eNOS) inhibitor, or anti-integrin alphaV antibody before culturing on HA surfaces with or without the presence of SNAP. Medium, cells alone or cells pretreated with these inhibitors or antibodies was used as the controls. Cell proliferation was assessed by colorimetric assay.
The results showed that SNAP, but not NAP, augmented HA-induced HOS cell proliferation. This modulatory effect of SNAP on HA-induced HOS cell proliferation was abolished by carboxy PTIO or anti-integrin alphaV antibody, but only partially reduced by L-NIO.
Therefore, the results of this study suggest that exogenous NO alone may up-regulate the proliferation of HOS cells attached on HA surfaces via integrin alphaV molecules.
本研究旨在验证外源性一氧化氮(NO)可能上调羟基磷灰石(HA)诱导的人成骨细胞系(HOS细胞)增殖这一假说。
将HOS细胞培养于HA表面,分别添加或不添加NO供体S-亚硝基乙酰青霉胺(SNAP)或亚硝基乙酰青霉胺(NAP)。在添加或不添加SNAP的细胞培养物中加入NO清除剂2-(4-羧基苯基)-4,4,5,5-四甲基咪唑啉-1-氧基-3-氧化物(羧基PTIO)。在将细胞培养于添加或不添加SNAP的HA表面之前,还用内皮型一氧化氮合酶(eNOS)抑制剂L-N5-(1-亚氨基乙基)鸟氨酸盐酸盐(L-NIO)或抗整合素αV抗体对细胞进行预处理。以培养基、单独的细胞或用这些抑制剂或抗体预处理的细胞作为对照。通过比色法评估细胞增殖。
结果显示,SNAP可增强HA诱导的HOS细胞增殖,而NAP则无此作用。羧基PTIO或抗整合素αV抗体可消除SNAP对HA诱导的HOS细胞增殖的这种调节作用,但L-NIO只能部分降低该作用。
因此,本研究结果表明,外源性NO单独作用时可能通过整合素αV分子上调附着于HA表面的HOS细胞的增殖。