氧化石墨烯-羟基磷灰石复合涂层材料的理化性质及生物相容性

Physicochemical properties and biocompatibility of graphene oxide-hydroxyapatite composite coating materials

背景:医用钛及钛合金种植体在临床应用中已经取得较好的治疗效果,但是仍存在种植体周围炎、松动、脱落等现象。目的:探究氧化石墨烯涂层材料的理化性能及其对骨髓间充质干细胞的影响。方法:①采用电化学沉积方法在钛片表面分别制备羟基磷灰石涂层和氧化石墨烯-羟基磷灰石复合涂层,通过扫描电镜、X射线能量色散谱、X射线光电子能谱、X射线衍射仪、拉曼光谱仪、接触角测量仪分析涂层的表面形貌、物相结构、官能团、元素成分及表面亲水性能;②分离培养小鼠骨髓间充质干细胞,分别接种于纯钛、羟基磷灰石涂层和氧化石墨烯-羟基磷灰石复合涂层表面,利用CCK-8法和扫描电镜对材料表面细胞的增殖、形态和生长状态进行评价。结果与结论:①扫描电镜、X射线能量色散谱显示,氧化石墨烯-羟基磷灰石复合涂层表面较羟基磷灰石涂层、纯钛更平展、致密及均匀;X射线光电子能谱、X射线衍射、拉曼光谱显示,在钛片表面成功制备了羟基磷灰石涂层、氧化石墨烯-羟基磷灰石复合涂层;氧化石墨烯-羟基磷灰石复合涂层的亲水性能较羟基磷灰石涂层、纯钛更优;②CCK-8检测显示,随着共培养时间的延长,3组材料表面的骨髓间充质干细胞数量增加,共培养1,4,7 d,氧化石墨烯-羟基磷灰石复合涂层表面的细胞增殖吸光度值均高于羟基磷灰石涂层、纯钛(P<0.0001);共培养4 d后的扫描电镜显示,3组材料表面的骨髓间充质干细胞均呈纺锤形,其中氧化石墨烯-羟基磷灰石复合涂层表面的细胞具有较好的拉伸性能,呈多边形,细胞边缘可见多个触角黏附于涂层表面,且表面的丝状足伸长量较纯钛和羟基磷灰石涂层多;③结果显示,氧化石墨烯-羟基磷灰石复合涂层材料是一种理化性能及生物学性能良好的涂层材料。

BACKGROUND:Medical titanium and titanium alloy have achieved good therapeutic effects in clinical applications,but there are still some phenomena such as peri-implant inflammation,loosening and shedding.OBJECTIVE:To explore the physicochemical properties of graphene oxide coating materials and their effects on bone marrow mesenchymal stem cells.METHODS:(1)Hydroxyapatite coating and graphene oxide/hydroxyapatite composite coating were prepared on a titanium surface by electrochemical deposition.The surface morphology,phase structure,functional groups,elemental composition and surface hydrophilicity of the coating were analyzed by scanning electron microscopy,X-ray energy dispersion spectroscopy,X-ray photoelectron spectroscopy,X-ray diffraction,Raman spectroscopy,and contact angle measurement instrument.(2)Mouse bone marrow mesenchymal stem cells were isolated and cultured and inoculated on pure titanium,hydroxyapatite coating and graphene oxide/hydroxyapatite composite coating,respectively.CCK-8 assay and scanning electron microscopy were used to evaluate the proliferation,morphology and growth status of the coated cells.RESULTS AND CONCLUSION:(1)Scanning electron microscopy and X-ray energy dispersion spectra showed that the surface of graphene oxide/hydroxyapatite composite coating was more flat,compact and uniform than that of hydroxyapatite coating and pure titanium.X-ray photoelectron spectroscopy,X-ray diffraction and Raman spectroscopy showed that hydroxyapatite coating and graphene oxide/hydroxyapatite composite coating were successfully prepared on the surface of the titanium sheet.The hydrophilicity of graphene oxide/hydroxyapatite composite coating was better than that of hydroxyapatite coating and pure titanium.(2)CCK-8 assay showed that the number of bone marrow mesenchymal stem cells on the surface of the three groups increased with the extension of co-culture time.On days 1,4,and 7,the proliferative absorbance of the cells on the graphene oxide/hydroxyapatite composite coating was higher than that on hydroxyapatite coating and pure titanium(P<0.0001).Scanning electron microscopy after 4 days of co-culture showed that bone marrow mesenchymal stem cells on the surface of the three groups of materials were fusiform.The cells on the surface of graphene oxide/hydroxyapatite composite coating showed good tensile properties and were polygonal.Multiple antennae could be seen attached to the coating surface at the edge of the cells,and the elongation of the filamentous foot on the surface was more than that on pure titanium and hydroxyapatite coatings.(3)The results show that graphene oxide/hydroxyapatite composite coating material is a kind of coating material with excellent physicochemical and biological properties.