不对称多孔抗菌nZnO/P34HB复合引导骨再生膜的制备及性能初步研究  

Preparation and properties of asymmetric porous antimicrobial nZnO/P34HB composite guided bone regeneration membrane

作　　者：刘畅[1] 刘为 罗婧美 韩莉 张超[1] 唐婉容[1] LIU Chang;LIU Wei;LUO Jingmei;HAN Li;ZHANG Chao;TANG Wanrong(Department of Stomatology,Affiliated Hospital of North Sichuan Medical College,Nanchong,Sichuan Province,637000,China)

机构地区：[1]川北医学院附属医院口腔科,四川南充637000

出　　处：《陆军军医大学学报》2024年第24期2745-2754,共10页Journal of Army Medical University

基　　金：川北医学院博士科研启动基金项目(CBY22-QDA11)。

摘　　要：目的制备不对称多孔抗菌纳米氧化锌(zinc oxide nanoparticle,nZnO)/聚3-羟基丁酸酯4-羟基丁酸酯[Poly(3-hydroxybutyrate-co-4-hydroxy butyrate)copolymer,P34HB]复合引导骨再生(guided bone regeneration,GBR)膜,并检测其成骨性能。方法本研究采用超声分散技术、溶剂置换法和冷冻干燥技术制备负载nZnO的P34HB复合GBR膜(实验分为空白对照组和0%、0.5%、1.0%、1.5%nZnO/P34HB材料组),采用扫描电子显微镜(scanning electron microscope,SEM)、能谱仪(energy dispersive spectrometer,EDS)等明确nZnO的负载并观察其结构;CCK-8检测细胞增殖能力;划痕实验检测细胞迁移能力;蛋白吸附实验检测GBR膜的蛋白吸附能力;酶解实验检测GBR膜的体外降解;成骨实验检测碱性磷酸酶(alkaline phosphatase,ALP)活性;茜素红染色观察钙化结节;体外抑菌实验检测GBR膜的抑菌能力。结果本研究成功制备了负载0%、0.5%、1.0%、1.5%nZnO的4组nZnO/P34HB复合GBR膜,SEM结果显示,该复合膜一面疏松多孔,另一面致密;EDS表明复合膜上出现锌元素的掺入。细胞增殖实验结果显示,0.5%、1.0%nZnO/P34HB组促进MC3T3-E1增殖且增殖数量均高于空白对照组、0%、1.5%nZnO/P34HB组(P<0.05);细胞划痕实验结果显示,在含有0.5%nZnO/P34HB组的浸提液培养基中MC3T3-E1明显越过划痕边缘进行迁移,且48 h时的0.5%nZnO/P34HB组细胞迁移率高于其他3个实验材料组(P<0.05);蛋白吸附实验结果显示,4组实验材料组光密度值差异无统计学意义,表明nZnO的加入不会降低材料本身的蛋白吸附能力;酶解实验结果显示,4组实验材料组的质量均随着酶解时间增加而持续降低,28 d后仍保持较低的降解率;ALP活性检测结果显示,0.5%nZnO/P34HB组ALP染色更深且活性高于其他3个实验材料组(P<0.01)。茜素红染色结果显示,0.5%nZnO/P34HB组与其他3个实验材料组比较,观察到较多的典型红色深染钙化结节,0%、0.5%nZnO/P34HB组的细胞矿化程度在4组中更Objective To develop an asymmetric porous guided bone regeneration(GBR)membrane made from antibacterial zinc oxide nanoparticle(nZnO)/poly(3-hydroxybutyrate-co-4-hydroxy butyrate)copolymer(P34HB),and detect its osteogenic performance.Methods Ultrasonic dispersion technique,solvent displacement method and freeze-drying technique were used to prepare P34HB composite GBR membrane loaded with nZnO at different doses(0%,0.5%,1.0%and 1.5%).Scanning electron microscopy(SEM)and energy spectrometry(EDS)were conducted to determine the load of nZnO and observe the structure of the prepared membrane.Then MC3T3-E1 cells were implanted in a 24-well culture plate with the prepared GBR membrane pre-laid in the well.Thus the experiment included blank control group(no membrane)and groups with membranes composing 0%,0.5%,1.0%and 1.5%nZnO,respectively.CCK-8 and cell scratch assays were used to detect cell proliferation capacity and migration ability.The protein adsorption capacity of GBR membrane was detected by protein adsorption experiment.The in vitro degradation of GBR membrane was tested;the activity of alkaline phosphatase(ALP)was detected with chemical test.And calcified nodules were observed with alizzarin red staining;the antibacterial ability of GBR membranes was tested with in vitro antibacterial experiments.Results Four groups of nZnO/P34HB composite GBR membranes with 0%,0.5%,1.0%and 1.5%nZnO were successfully prepared.SEM results showed that the composite membrane was porous on one side and dense on the other side.EDS indicated the incorporation of zinc elements in the composite membrane.The results of cell proliferation assay showed that 0.5%and 1.0%nZnO/P34HB group promoted the proliferation of MC3T3-E1 cells,when compared with the blank control group and 0%and 1.5%nZnO/P34HB groups(P<0.05).The results of cell scratch assay showed that MC3T3-E1 cells in the 0.5%nZnO/P34HB group migrated significantly over the scratch edge,with the cell mobility higher than that of the other 3 membrane groups in 48 h after culture(P<0

关 键 词：不对称多孔结构 径向排列纤维 纳米氧化锌 生物可降解材料 引导性骨再生 

分 类 号：R318.08[医药卫生—生物医学工程] R782.13[医药卫生—基础医学] R944.9