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碘涂层骨科钛内植物的制备及抗菌性能的评估

摘要

目的 通过电泳沉积法在钛克氏针上进行碘的负载,并验证其抗菌性能.方法 采用电泳沉积法对钛克氏针表面进行碘的负载以制备碘涂层钛克氏针,分别配制1 000 ppm、2 000 ppm、4 000 ppm聚维酮碘水溶液(l ppm=1 mg/kg=1 mg/L)为电解液,经抛光打磨后的钛克氏针作为阳极,铂片作为阴极,在200V的电压下电泳沉积30 min,再用蒸馏水进行超声清洗后烘干,分别制得3种含碘量不同的碘涂层骨科钛克氏针.通过扫描电镜及能谱分析,研究碘涂层钛克氏针的表面征象及成分结构.体外抗菌实验分组:未涂层钛克氏针10根对照组,1 000 ppm碘涂层钛克氏针10根,2000 ppm碘涂层钛克氏针10根,4 000 ppm碘涂层钛克氏针为10根.分别将钛针浸没在106 CFU/ml的金黄色葡萄球菌(ATCC25923)菌悬液中,以便金葡菌充分的接种在钛针表面.在37°C下细菌培养箱中放置6h后取出,再利用超声震荡将生长在钛针表面的细菌震荡下来,稀释10000倍后取100μl菌液涂抹在琼脂培养基中进行培养(37℃),24h后观察、计数菌落数量.结果 电泳沉积法制备的3种碘含量不同的碘涂层钛克氏针外观呈褐色,表面覆盖均匀的含碘化合物涂层,膜层结构稳定,超声清洗后无脱落.电镜下观察可见钛克氏针表面大致均匀覆盖一层较为致密的涂层,外观结构较为平整,并可见散在分布些许轻度陷凹,能谱仪分析结果提示1 000 ppm组、2 000 ppm组、4 000 ppm组的表面涂层碘含量依次为4.38 wt%、9.05 wt%、14.48wt%.利用超声震荡将生长在钛针表面的细菌震荡下来,其菌液经稀释、涂布、培养24h后行CFU计数,可见随着碘涂层含量的上升,其抗菌性能不断的提升,差异有统计学意义;其中含碘量14.48 wt%的碘涂层内植物的抗菌性能超过对照组1个数量级以上.结论 利用电泳沉积法制备碘涂层钛针,经电镜扫描观测表征、能谱仪分析元素结构、定量测定,证明碘的成功负载;体外抗菌性能评估证明,在实验室条件下碘涂层钛针的抗菌性能优于普通钛针.%Objective Iodine loading would be carried out on orthopedic titanium Kirschner-wire by electrophoretic deposition.And by this,a kind of orthopedic iodine-coated implant with antibacterial properties was supposed to be developed,that would provide a theoretical basis for the development of clinical orthopedic implants with antibacterial properties.Methods Iodine loading on the surface of titanium Kirschner-wire was carried out by electrophoretic deposition.PVP-I solution as 1 000 ppm,2 000 ppm and 4 000 ppm was prepared respectively (1 ppm=1 mg/kg=1 mg/L).Three kinds of iodine-coated orthopedic titanium Kirschner-wires with different iodine content were prepared by electrophoretic deposition at 200 V voltage for 30 min and then stoved after being washed with distilled water.The surface signs and composition structure of iodine coated titanium Kirschnerwire were studied through scanning electron microscopy (SEM) and energy dispersive spectrometer (EDS).Antibacterial experiments of Iodine coating titanium Kirschner needle were performed in vitro.Antimicrobial test:10 pieces of uncoated titanium Kirschner-wires;10 pieces of iodine-coated titanium Kirschner-wires with 1 000 ppm;10 pieces of iodine-coated titanium Kirschner-wires with 2 000 ppm;10 pieces of iodine-coated titanium wires with 4 000 ppm.Because iodine is easy to sublimate and does not withstand high temperature,all the titanium Kirschner needles are fumigated and disinfected by ethylene oxide (supported by the supply room of our hospital).Titanium wires were immersed respectively in 106 CFUJ/ml (ATCC25923) bacterial suspension of standard Staphylococcus aureus so that bacteria would be fully inoculated on the surface of the titanium wire.The sterilized titanium Kirschner needle was placed in a sterile culture cup and 30 ml suspension was added into the culture cup.The sterilized titanium Kirschner needle was completely immersed in the bacterial solutionand was kept at 37°C for 6 h before taking out.Take out the titanium Kirschner needle slowly,rinse the titanium Kirschner needle with 5 ml sterile PBS buffer,after rinsing the surface of titanium Kirschner needle,transfer the titanium Kirschner needle to the new sterile culture cup and add 5 ml sterile saline until the titanium Kirschner needle is completely immersed.Then put the titanium Kirschner needle into the ultrasonic oscillator to wash the surface of the bacteria fully.After 10 000 times dilution,100 μl was smeared on agar medium for culture (37°C) and the number of colonies was observed and counted 24 h later.Results The surface of three kinds of iodine coating titanium Kirschner-wires was covered with uniform iodized compound coating and had a brown look.The film structure was stable and there was no falling off after ultrasonic cleaning.SEM exhibited that the surface of Titanium Kirschner-wire is covered with a compact coating;the appearance structure is relatively flat and a slight collapse can be seen scattered in the surface of the wire.The results of EDS showed that the iodine content in the surface coating of 3 groups was 4.38 wt%,9.05 wt% and 14.48 wt%.The bacteria growing on the surface of titanium needle were shaken down by ultrasonic vibration.The CFU counting was carried out after the bacterial solution being diluted,coated and cultured for 24 h.It can be seen that the antibacterial property increases continuously with the increase of iodine coating content.The antibacterial properties of iodine-coated implant which content 14.48 wt% iodine was more than 1 orders of magnitude than the control group.The results showed that the plants coated with iodine had obvious antibacterial effect.Conclusion The iodine coated titanium wire was prepared by electrophoretic deposition,after being characterized by SEM,EDS and quantitative measurement,the successful loading of iodine was proved,the antibacterial experiment proved that the iodinecoated orthopedic titanium Kirschner-wire has a stronger antibacterial properties than the ordinary titanium Kirschner-wire under laboratory conditions.

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