经多巴胺或硅烷化修饰的氧化石墨烯钛表面涂层的体外成骨性能研究

doi:10.7518/hxkq.2025.2024357

华西口腔医学杂志 ›› 2025, Vol. 43 ›› Issue (3): 336-345.doi: 10.7518/hxkq.2025.2024357

经多巴胺或硅烷化修饰的氧化石墨烯钛表面涂层的体外成骨性能研究

吴清霖(), 赖颖真(), 黄艳玲, 谢泽宇, 林彦吟

厦门医学院口腔医学院口腔生物材料福建省高校工程研究中心，厦门 361023

收稿日期:2024-09-25 修回日期:2024-11-29 出版日期:2025-06-01 发布日期:2025-06-10
通讯作者: 赖颖真 E-mail:1976717712@qq.com;dentistyz@126.com
作者简介:吴清霖，学士，E-mail：1976717712@qq.com
基金资助:
福建省自然科学基金(2022J011408);厦门市医疗卫生指导性项目(3502Z20244ZD1134);厦门医学院校级科研项目(K2023-01)

In vitro osteogenic performance study of graphene oxide-coated titanium surfaces modified with dopamine or silane

Wu Qinglin(), Lai Yingzhen(), Huang Yanling, Xie Zeyu, Lin Yanyin

School of Stomatology, Xiamen Medical College & Engineering Research Center of Stomatological Biomaterials, Fujian Province University, Xiamen 361023, China

Received:2024-09-25 Revised:2024-11-29 Online:2025-06-01 Published:2025-06-10
Contact: Lai Yingzhen E-mail:1976717712@qq.com;dentistyz@126.com
Supported by:
Fujian Natural Science Foundation Project(2022J011408);Xiamen City Medical and Health Guiding Project(3502Z20244ZD1134);Scientific Research Project of Xiamen Medical College(K2023-01)

摘要/Abstract

摘要：

目的本研究通过比较经多巴胺或硅烷化修饰的氧化石墨烯钛表面涂层的成骨性能差异，旨在尝试为钛表面氧化石墨烯涂层的修饰提供较为适宜的方案。方法钛碱热处理后分别进行多巴胺和硅烷化改性，被覆氧化石墨烯涂层，设计对照组：纯钛（Ti）组；实验组：钛经碱热处理（Ti-NaOH）组、钛经碱热处理后硅烷化改性（Ti-APTES）组、钛经碱热处理后多巴胺改性（Ti-DOPA）组、钛经硅烷化改性表面修饰氧化石墨烯（Ti-APTES/GO）组、钛经多巴胺改性表面修饰氧化石墨烯（Ti-DOPA/GO）组。利用扫描电子显微镜、接触角测量仪、X射线光电子能谱分析仪及拉曼光谱分析仪对材料表面的理化性能进行分析。通过细胞活性检测、免疫荧光染色后激光共聚焦显微镜观察小鼠胚胎成骨细胞前体细胞MC3T3-E1在材料表面的增殖及黏附形态。通过碱性磷酸酶、茜素红染色及定量、实时荧光定量聚合酶链反应研究对MC3T3-E1细胞成骨分化的影响。结果氧化石墨烯涂层修饰后扫描电子显微镜观察可见覆盖一层薄膜样结构，实验组亲水性皆有改善，其中Ti-DOPA/GO组亲水性最佳。X射线光电子能谱学和拉曼光谱分析显示氧化石墨烯修饰后的材料均出现典型D峰和G峰，表面出现大量含氧官能团。细胞计数盒检测可见各组材料均无细胞毒性，Ti-APTES/GO组增殖水平高于Ti-DOPA/GO组。激光共聚焦显微镜下可见Ti-DOPA/GO组和Ti-APTES/GO组细胞铺展更为饱满。Ti-DOPA/GO组与Ti-APTES/GO组碱性磷酸酶染色最深，Ti-APTES/GO组茜素红染色矿化结节最多，茜素红染色定量结果最高；在Ti-DOPA/GO组和Ti-APTES/GO组中，早期成骨相关基因RUNX2的表达均达到较高水平，而在晚期成骨相关基因OPN和OCN的表达上，Ti-APTES/GO组的表现优于Ti-DOPA/GO组。结论 Ti-APTES/GO在促进MC3T3-E1细胞的黏附、增殖及体外成骨分化方面，效果显著优于Ti-DOPA/GO。

关键词: 钛, 氧化石墨烯, 表面改性, 成骨分化

Abstract:

Objective This study aimed to compare the osteogenic performance differences of titanium surface coatings modified by dopamine or silanized graphene oxide, and to provide a more suitable modification scheme for titanium surface graphene oxide coatings. Methods Titanium was subjected to alkali-heat treatment and then modified with dopamine and silanization, respectively, followed by coating with graphene oxide. Control and experimental groups were designed as follows: pure titanium (Ti) group; titanium after alkali-heat treatment (Ti-NaOH) group; titanium after alkali-heat treatment and silanization modification (Ti-APTES) group; titanium after alkali-heat treatment and dopamine modification (Ti-DOPA) group; titanium with silanization-modified surface decorated with graphene oxide (Ti-APTES/GO) group; titanium with dopamine-modified surface decorated with graphene oxide (Ti-DOPA/GO) group. The physical and chemical properties of the material surfaces were analyzed using scanning electron microscopy (SEM), contact angle goniometer, X-ray photoelectron spectroscopy (XPS), and Raman spectrometer. The proliferation and adhesion morphology of mouse embryonic osteoblast precursor cells MC3T3-E1 on the material surfaces were observed by cell viability detection and immunofluorescence staining followed by laser confocal microscopy. The effects on the osteogenic differentiation of MC3T3-E1 cells were studied by alkaline phosphatase (ALP) staining, alizarin red staining and quantification, and real-time quantitative polymerase chain reaction. Results After modification with graphene oxide coating, a thin-film-like structure was observed on the surface under SEM. The hydrophilicity of all experimental groups was improved, among which the Ti-DOPA/GO group had the best hydrophilicity. XPS and Raman spectroscopy analysis showed that the modified materials exhibited typical D and G peaks, and XPS revealed the presence of a large number of oxygen-containing functional groups on the surface. CCK8 assay showed that all groups of materials had no cytotoxicity, and the proliferation level of the Ti-APTES/GO group was higher than that of the Ti-DOPA/GO group. Under the laser confocal microscope, the cells in the Ti-DOPA/GO and Ti-APTES/GO groups spread more fully. The Ti-DOPA/GO and Ti-APTES/GO groups had the deepest ALP staining, and the Ti-APTES/GO group had the most alizarin red-stained mineralized nodules and the highest quantitative result of alizarin red staining. In the Ti-DOPA/GO and Ti-APTES/GO groups, the expression of the early osteogenic-related gene RUNX2 reached a relatively high level, while in the expression of the late osteogenic-related genes OPN and OCN, the Ti-APTES/GO group performed better than the Ti-DOPA/GO group. Conclusion Ti-APTES/GO significantly outperformed Ti-DOPA/GO in promoting the adhesion, proliferation, and in vitro osteogenic differentiation of MC3T3-E1 cells

Key words: titanium, graphene oxide, surface modification, osteogenic differentiation

中图分类号:

R782

吴清霖, 赖颖真, 黄艳玲, 谢泽宇, 林彦吟. 经多巴胺或硅烷化修饰的氧化石墨烯钛表面涂层的体外成骨性能研究[J]. 华西口腔医学杂志, 2025, 43(3): 336-345.

Wu Qinglin, Lai Yingzhen, Huang Yanling, Xie Zeyu, Lin Yanyin. In vitro osteogenic performance study of graphene oxide-coated titanium surfaces modified with dopamine or silane[J]. West China Journal of Stomatology, 2025, 43(3): 336-345.

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基因	引物序列（5'-3'）
RUNX2	AACGATCTGAGATTTGTGGGC CCTGCGTGGGATTTCTTGGTT
ALP	CCAACTCTTTTGTGCCAGAGA GGCTACATTGGTGTTGAGCTTTT
COL-I	GCTCCTCTTAGGGGCCACT CCACGTCTCACCATTGGGG
OCN	CTGACCTCACAGATCCCAAGC TGGTCTGATAGCTCGTCACAAG
OPN	AGCAAGAAACTCTTCCAAGCAA GTGAGATTCGTCAGATTCATCCG
Osterix	ACCCCAAGATGTCTATAAGCCC CGCTCTAGCTCCTGACAGTTG
GAPDH	ACAACTTTGGTATCGTGGAAGG GCCATCACGCCACAGTTTC

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经多巴胺或硅烷化修饰的氧化石墨烯钛表面涂层的体外成骨性能研究

In vitro osteogenic performance study of graphene oxide-coated titanium surfaces modified with dopamine or silane

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