Kim Ga-Hyun, Kim Il-Shin, Park Sang-Won, Lee Kwangmin, Yun Kwi-Dug, Kim Hyun-Seung, Oh Gye-Jeong, Ji Min-Kyung, Lim Hyun-Pil
J Nanosci Nanotechnol. 2016 Feb;16(2):1396-9. doi: 10.1166/jnn.2016.11916.
This study examined the effect of the immobilization of the Gly-Arg-Gly-Asp-Ser (GRGDS) peptide on titanium dioxide (TiO2) nanotube via chemical grafting on osteoblast-like cell (MG-63) viability and differentiation. The specimens were divided into two groups; TiO2 nanotubes and GRGDS-immobilized TiO2 nanotubes. The surface characteristics of GRGDS-immobilized TiO2 nanotubes were observed by using X-ray photoelectron spectroscopy (XPS) and a field emission scanning electron microscope (FE-SEM). The morphology of cells on specimens was observed by FE-SEM after 2 hr and 24 hr. The level of cell viability was investigated via a tetrazolium (XTT) assay after 2 and 4 days. Alkaline phosphatase (ALP) activity was evaluated to measure the cell differentiation after 4 and 7 days. The presence of nitrogen up-regulation or C==O carbons con- firmed that TiO2 nanotubes were immobilized with GRGDS peptides. Cell adhesion was enhanced on the GRGDS-immobilized TiO2 nanotubes compared to TiO2 nanotubes. Furthermore, significantly increased cell spreading and proliferation were observed with the cells grown on GRGDS-immobilized TiO2 nanotubes (P < .05). However, there was no significant difference in ALP activity between GRGDS-immobilized TiO2 nanotubes and TiO2 nanotubes. These results suggest that the GRGDS-immobilized TiO2 nanotubes might be effective in improving the osseointegration of dental implants.
本研究通过化学接枝法将甘氨酸-精氨酸-甘氨酸-天冬氨酸-丝氨酸(GRGDS)肽固定在二氧化钛(TiO₂)纳米管上,考察其对成骨样细胞(MG-63)活力和分化的影响。将标本分为两组:TiO₂纳米管组和GRGDS固定化TiO₂纳米管组。采用X射线光电子能谱(XPS)和场发射扫描电子显微镜(FE-SEM)观察GRGDS固定化TiO₂纳米管的表面特性。在2小时和24小时后,通过FE-SEM观察标本上细胞的形态。在2天和4天后,通过四氮唑(XTT)法研究细胞活力水平。在4天和7天后,评估碱性磷酸酶(ALP)活性以测量细胞分化情况。氮上调或C==O碳的存在证实了TiO₂纳米管已固定有GRGDS肽。与TiO₂纳米管相比,GRGDS固定化TiO₂纳米管上的细胞黏附增强。此外,观察到在GRGDS固定化TiO₂纳米管上生长的细胞的铺展和增殖显著增加(P <.05)。然而,GRGDS固定化TiO₂纳米管和TiO₂纳米管之间的ALP活性没有显著差异。这些结果表明,GRGDS固定化TiO₂纳米管可能在改善牙种植体的骨整合方面有效。