构建淫羊藿苷/聚己内酯纳米纤维膜及体外促成骨能力

Construction of icariin/polycaprolactone nanofiber membrane and its osteogenic ability in vitro

背景:在骨组织工程中,静电纺丝的应用使得促成骨生长因子能够更长久的发挥作用。研究表明,淫羊藿苷可促进MC3T3-E1细胞的增殖。目的:构建载淫羊藿苷/聚己内酯纳米纤维膜,评估其生物相容性及体外促成骨能力。方法:采用同轴静电纺丝技术制备淫羊藿苷/聚己内酯纳米纤维膜,表征其微观形貌及体外药物释放。体外培养MC3T3-E1细胞,分4组处理:空白组常规培养细胞,纳米纤维膜组加入聚己内酯纳米纤维膜,药物组加入含淫羊藿苷的培养基,实验组加入淫羊藿苷/聚己内酯纳米纤维膜,检测各组细胞骨架形态、增殖、碱性磷酸酶活性及骨钙素mRNA表达。结果与结论:①扫描电镜显示,淫羊藿苷/聚己内酯纳米纤维膜具有三维立体的网状交织的结构,相互交错,空隙大小均一,孔径100-200μm,纤维平均分布范围为300-600 nm;②观察淫羊藿苷/聚己内酯纳米纤维膜1个月内的药物释放累积量,开始1-5 d药物有突释现象,释药量达45%左右,后期药物呈缓慢持续释放,到30 d时药物释放量达80%以上;③培养24 h后激光共聚焦显微镜下可见,细胞黏附于纳米纤维膜生长,细胞肌动蛋白走行及细胞内部结构清晰;④CCK-8检测显示,实验组培养4,7 d的细胞增殖快于其他3组(P<0.05);⑤实验组诱导培养7,14,21 d的碱性磷酸酶活性均高于其他3组(P<0.05),诱导培养7 d的骨钙素mRNA表达高于其他3组(P<0.05);⑥结果表明,采用静电纺丝法可构建淫羊藿苷/聚己内酯纳米纤维膜,该膜具有良好的体外释药性能与促成骨性能。

BACKGROUND:In bone tissue engineering,the application of electrospinning makes bone growth factor play a long-term role.Icariin has been shown to promote the proliferation of MC3T3-E1 cells.OBJECTIVE:To construct icariin/polycaprolactone nanofiber membrane,and evaluate its biocompatibility and osteogenic ability in vitro.METHODS:Coaxial electrospinning technology was used to prepare icariin/polycaprolactone nanofiber membrane,and their morphology and drug release in vitro were characterized.MC3T3-E1 cells were cultured in vitro and divided into four groups.The blank group was treated with conventional culture.The nanofiber membrane group was treated with polycaprolactone nanofiber membrane.The drug group was treated with medium containing icariin.The experimental group was treated with icariin/polycaprolactone nanofiber membrane.Cytoskeleton morphology,proliferation,alkaline phosphatase activity,and osteocalcin mRNA expression were detected in each group.RESULTS AND CONCLUSION:(1)Scanning electron microscope revealed that the icariin-loaded/polycaprolactone electrospinning nanofiber membrane showed a three-dimensional structure,uniform void size,aperture of 100-200μm,distribution range of 300-600 nm.(2)The cumulative drug release of icariin/polycaprolactone nanofiber membrane observed within one month demonstrated that the drug release was about 45%in the first 1-5 days.The drug release was slow and sustained in the later period,and more than 80%in the 30 days.(3)After 24 hours of culture,laser confocal microscopy showed that the cells adhered to the nanofiber membrane,and the actin and internal structure of the cells were clear.(4)CCK-8 assay exhibited that the cell proliferation of the experimental group was faster than that of the other three groups at 4 and 7 days(P<0.05).(5)Alkaline phosphatase activity was higher in the experimental group than that in the other three groups at 7,14,and 21 days(P<0.05).Osteocalcin mRNA expression was higher in the experimental group than that in the other three groups at 7 days after culture(P<0.05).(6)The results show that electrospinning can be used to construct icariin/polycaprolactone nanofiber membrane,which has good drug release and bone promoting properties in vitro.