载白藜芦醇及BMP-2同轴纳米纤维膜的构建及其在牙周炎骨缺损重建中应用的实验研究

Experimental Study of Fabrication of Resveratrol and BMP-2 Loaded Coaxial Nanofiber Membrane and its Application in the Reconstruction of Periodontitis Bone Defects

目的:制备载白藜芦醇(RES)及骨形态发生蛋白-2(BMP-2)的同轴纳米纤维膜,初步探究其对牙周组织炎症的抑制作用及其对根分叉病变牙槽骨缺损的修复作用。方法:通过同轴静电纺丝技术制备具有芯-壳结构的纳米纤维膜,将白藜芦醇负载于纳米纤维的壳层,BMP-2负载于芯层。采用透射电镜、扫描电镜等对其进行表征;采用CCK-8法及qRT-PCR技术体外初步探讨其生物相容性、炎症抑制作用及促进干细胞成骨分化的作用。构建比格犬II度根分叉病变模型,并探究所制备的载药纳米纤维膜对牙周缺损组织的修复作用。结果:本实验制备的同轴纳米纤维膜,平均直径约为(280±12)nm,其表现出明显的芯-壳结构;其具有良好的生物相容性,同时qRT-PCR结果显示同轴纳米纤维膜可显著下调牙周膜细胞炎症因子表达(P<0.05),并促进骨髓间充质干细胞成骨相关基因表达(P<0.05)。动物实验显示,载白藜芦醇及BMP-2的同轴纳米纤维膜可以显著促进根分叉缺损的骨再生。结论:载白藜芦醇及BMP-2的同轴纳米纤维膜,可抑制牙周炎相关因子的表达,并促进成骨基因的表达,其对比格犬II度根分叉病变骨缺损具有一定促进修复作用。

Objective:A coaxial nanofibric membrane containing resveratrol and BMP-2 was prepared to investigate its inhibitory effect on inflammatory factors.To explore the repair effect on the alveolar bone defect in the II degree bifurcation periodontitis disease model of the Beagle dog.Methods:A nanofiber membrane with a core-shell structure was prepared by coaxial electrospinning technique.Resveratrol was loaded into the nanofiber shell,and the BMP-2 was loaded into the core layer.The morphology and structure of nanofibers were detected by transmission electron microscopy(TEM)and scanning electron microscopy(SEM)respectively.The effect of coaxial nanofiber membranes on cell proliferation was detected by CCK-8 assay.qRT-PCR was used to detect the effects of expression of inflammatory cytokines and its effect on stem cells osteogenic gene expression.Beagle II-degree bifurcation lesion model was constructed to study its repairing effect on the bone defect.Results:The average diameter of the coaxial nanofibers prepared in this experiment was about(280±12)nm,which showed a clear core-shell structure.The periodontal ligament cells could be well proliferated on the surface of the coaxial nanofiber membrane,The results of qRT-PCR showed that the coaxial nanofiber membranes decreased the expression of inflammatory cytokines(P<0.05).It also intensified the expression of osteogenic genes of bone marrow mesenchymal stem cells(BMSCs)(P<0.05).The results of animal experiments showed that the coaxial nanofiber membranes containing resveratrol and BMP-2 could significantly promote bone regeneration in Beagle dogs with II-degree furcation defects.Conclusion:In this study,coaxial nanofiber membranes containing resveratrol and BMP-2 were successfully prepared,and it was confirmed that the nanofiber membranes can inhibit the expression of periodontitis related fac-tors and promote the expression of osteogenic genes.The coaxial nanofiber membrane has a repairing effect on the bone defect of the dog II degree furcation lesions,and is expected to be applied in the regeneration of periodontal bone defects.