抗牙龈卟啉单胞菌卵黄抗体脂质体的制备及抗菌作用研究

Study on the inhibitory effect of anti-P.gingivalis yolk antibody liposome on periodontal pathogenic bacteria

目的通过制备特异性抗牙龈卟啉单胞菌卵黄抗体脂质体(抗Pg-IgY脂质体),探讨特异性抗Pg-IgY脂质体对牙周主要致病菌牙龈卟啉单胞菌体外抑菌效果。方法采用薄膜分散法制备抗Pg-IgY脂质体。使用透射电子显微镜观察抗Pg-IgY脂质体外观表征,纳米粒径分析仪测定抗Pg-IgY脂质体的Zeta电位(ζ-电位)和粒径,利用超滤离心法测量其包封率。采用微量液体稀释法研究抗Pg-IgY脂质体对牙龈卟啉单胞菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC)并绘制其kill-time曲线。结果测得抗Pg-IgY脂质体平均粒径为(175.43±3.25)nm(n=4)、多分散指数为0.263(n=4)、Zeta电位为(-37.82±0.20)mV(n=4)。抗Pg-IgY脂质体包封率为42.71%,药物累积释放量为83.4%。抗Pg-IgY脂质体对牙龈卟啉单胞菌MIC值为4 mg/ml,MBC值为8 mg/ml。当达到细菌的MBC值可完全抑制细菌在培养液中的生长繁殖。结论抗Pg-IgY脂质体对牙龈卟啉单胞菌具有良好的抑制作用,达到最小的杀菌浓度时可以杀灭细菌。

Objective To prepare liposomes of egg yolk antibodies against Porphyromonas gingivalis(anti-Pg-IgY liposome),and to study the antibacterial effect of these anti-Pg-IgY liposomes on Porphyromonas gingivalis,a major periodontal pathogen.Methods Anti-Pg-IgY liposomes were preparedby thin film dispersion.Use transmission electron microscope to observe the appearance and characterization of anti-Pg-IgY liposomes.Zeta potential(ζ-potential)and particle size of anti-Pg-IgY liposomes was determined by ultrafiltration centrifugation method.The micro-liquid dilution method was used to study the minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of anti-Pg-IgY liposomes against Porphyromonas gingivalis and draw its kill-time curve.Results The average particle size of the liposomes was(175.43±3.25)nm(n=4),while the polydispersity index was 0.263(n=4),and the zeta potential was(-37.82±0.20)mV(n=4).The encapsulation efficiency was 42.71%(n=4),and the cumulative release rate was 83.4%.The MIC and MBC values of the Pg-Igy liposomes against porphyromonas gingivalis were 4 mg/ml and 8 mg/ml,respectively.When the anti-Pg-IgY liposome reached the MBC value of bacteria,it could completely inhibit the growth and reproduction of bacteria in the culture medium.Conclusion The anti-Pg-IgY liposomes exhibit excellent antibacterial activity,and can kill the bacteria when the minimum germicidal concentration is reached.