牙龈卟啉单胞菌对牙髓成纤维细胞胞内感染的研究  

Establishment of a model of intracellular infection on human dental pulp fibroblast by Porphyromonas gingivalisin vitro

作　　者：卢煜[1] 罗世高[1] 黄定明[1] 

机构地区：[1]口腔疾病研究国家重点实验室,四川大学,四川成都610041

出　　处：《华西口腔医学杂志》2008年第1期87-89,93,共4页West China Journal of Stomatology

基　　金：国家自然科学基金资助项目(30572034)

摘　　要：目的研究牙龈卟啉单胞菌活菌细胞内感染牙髓成纤维细胞的体外模型。方法将牙龈卟啉单胞菌和牙髓细胞以100∶1比例共同孵育0.5、1.0、2.0h,倒置显微镜观察牙髓细胞形态。加入双抗和甲硝唑杀死胞外细菌,牙髓细胞用蒸馏水裂解后厌氧培养细胞裂解液,观察活细菌是否进入牙髓细胞胞内。将牙龈卟啉单胞菌和牙髓细胞以多重感染比(MOI)100、50共同孵育1.0h,采用MTT法检测感染牙龈卟啉单胞菌后牙髓细胞存活率。结果倒置显微镜下显示,被胞内感染的牙髓细胞培养2.0h未见胞膜破裂,形态完整。采用双抗能彻底杀灭胞外培养基中的细菌而对胞内细菌无影响,共同孵育1.0h和2.0h活细菌能进入牙髓细胞。MTT法显示细菌感染后牙髓细胞仍有一定存活率,其存活率分别是MOI100组74.43%、MOI50组99.07%。结论成功建立了牙龈卟啉单胞菌对牙髓成纤维细胞胞内感染的模型。Objective To establish a model of intracellular infection of human dental pulp fibroblast （HDPF） by Porphyromonas gingivalis in vitro. Methods HDPF was incubated with live Porphyromonas gingivalis with a multiplicitas of infection （MOI） from 100 for 0.5, 1.0, 2.0 hours. HDPF infected with live Porphyromonas gingivalis were observed by inverted microscope. Metronidazole, Penicillin, Streptomycin were added to the medium to kill any extracellular bacteria. Then the cells were lysed by sterile distilled water, dilutions of the lysates of cells were cultured anaerobically. Colony of invasive bacteria were then observed. HDPF exposed live Porphyromonas gingivalis with a MOI 50 and 100 for 1.0 hour, then cells survival rate were detected by MTT assay. Results After infected with Porphyromonas gingivalis 2.0 hours, cells were eumorphism. Porphyromonas gingivalis of incubation for 1.0, 2.0 hours was invasive cells. MTT assays indicated 99.07% cells survival rate with a MOI from 50 and 74.43% cells survival rate with a MOI from 100. Conclusion A model of HDPF intracellular infection with Porphyromonas gingivalis was successfully established in vitro.

关 键 词：牙龈卟啉单胞菌 牙髓成纤维细胞: 胞内感染 

分 类 号：R78[医药卫生—口腔医学]