静电纺丝蛋清/聚乙烯醇/氧化石墨烯复合纤维支架的制备及体外生物相容性

Preparation and in vitro biocompatibility of egg white/polyvinyl alcohol/graphene oxide composite fiber scaffolds based on electrospinning

背景:相关研究表明,添加适量的氧化石墨烯可以改善纤维支架的机械性能、亲水性和细胞相容性,并具有一定的促进成骨性能。目的:制备含不同质量浓度氧化石墨烯的蛋清/聚乙烯醇/氧化石墨烯复合纤维支架,初步探索该复合纤维支架的体外生物相容性。方法:采用静电纺丝技术制备含氧化石墨烯0,0.5,1,1.5 g/L的蛋清/聚乙烯醇/氧化石墨烯复合纤维支架,扫描电镜观察支架形貌,同时检测支架的机械性能、接触角与平衡溶胀比。将人脐静脉血管内皮细胞和人骨髓间充质干细胞分别接种于4组支架表面,采用CCK-8法检测细胞增殖,Live/Dead荧光染色观察细胞活性,划痕实验和罗丹明鬼笔环肽染色观察人脐静脉血管内皮细胞的迁移与形态。结果与结论:①扫描电镜显示,随着氧化石墨烯含量的增加,复合支架的纤维直径增加;②随着氧化石墨烯含量的增加,复合支架的亲水性先降低后升高,其中含氧化石墨烯0.5 g/L复合支架的亲水性最低;③随着氧化石墨烯含量的增加,复合支架的杨氏模量和拉伸强度先增大后减小,其中含氧化石墨烯0.5 g/L复合支架的杨氏模量和拉伸强度最大;④CCK-8实验显示,含氧化石墨烯1.5 g/L复合支架上人骨髓间充质干细胞的增殖速率快于其他3组支架(P<0.05),含氧化石墨烯1 g/L复合支架上人脐静脉血管内皮细胞的增殖速率快于其他3组支架(P<0.05);⑤随着氧化石墨烯含量的增加,复合支架上人骨髓间充质干细胞的存活率增加;含氧化石墨烯1 g/L复合支架上人脐静脉血管内皮细胞的存活率最高;⑥划痕实验和罗丹明鬼笔环肽染色证明,含氧化石墨烯1 g/L复合支架最有利于人脐静脉血管内皮细胞的迁移和形态扩张;⑦结果表明,含氧化石墨烯1 g/L的蛋清/聚乙烯醇/氧化石墨烯纤维支架具有优良的纤维结构、亲水性、力学性能及细胞相容性。

BACKGROUND:Relevant studies have shown that adding an appropriate amount of graphene oxide can improve the mechanical properties,hydrophilicity,and cell compatibility of the fiber scaffold,and has a certain ability to promote bone formation.OBJECTIVE:To prepare egg white/polyvinyl alcohol/graphene oxide composite fiber scaffolds with different concentrations of graphene oxide and preliminary explore the biocompatibility of composite fiber scaffold in vitro.METHODS:Electrospinning was used to prepare egg white/polyvinyl alcohol/graphene oxide composite fiber scaffolds containing 0,0.5,1,and 1.5 g/L graphene oxid.The morphology of the scaffold was observed by scanning electron microscope,and the mechanical properties,contact angle and equilibrium swelling ratio of the scaffold were also tested.Human umbilical vein endothelial cells and human bone marrow mesenchymal stem cells were respectively inoculated on the surface of four groups of scaffolds.The CCK-8 assay was used to detect the proliferation of the two types of cells.Live/Dead fluorescent staining was used to observe the viability of the two types of cells.The scratch test and rhodamine phalloidin staining were used to observe the migration and morphology of human umbilical vein endothelial cells.RESULTS AND CONCLUSION:(1)Scanning electron microscope showed that with the increase of graphene oxid content,the fiber diameter of the composite scaffold increased.(2)With the increase of graphene oxid content,the hydrophilicity of the composite scaffold first decreased and then increased,and the hydrophilicity of the composite scaffold containing 0.5 g/L graphene oxid was the lowest.(3)With the increase of graphene oxid content,the Young's modulus and tensile strength of the composite scaffold first increased and then decreased,and the Young's modulus and tensile strength of the composite scaffold containing 0.5 g/L graphene oxid were the largest.(4)CCK-8 assay showed that the proliferation rate of human bone marrow mesenchymal stem cells on the composite scaffold containing 1.5 g/L graphene oxid was faster than that of the other three groups of scaffold(P<0.05),and the proliferation rate of human umbilical vein endothelial cells on the composite scaffold containing 1 g/L graphene oxid was faster than the other three groups of scaffolds(P<0.05).(5)With the increase of graphene oxid content,the survival rate of human bone marrow mesenchymal stem cells on the composite scaffold increased;the survival rate of human umbilical vein endothelial cells on the composite scaffold containing 1 g/L graphene oxid was the highest.(6)Scratch experiment and rhodamine phalloidin staining proved that the composite scaffold containing 1 g/L graphene oxid was most conducive to the migration and morphological expansion of human umbilical vein endothelial cells.(7)The results show that the egg white/polyvinyl alcohol/graphene oxide fiber scaffold containing 1 g/L graphene oxid has excellent fiber structure,hydrophilicity,mechanical properties and cell compatibility.