髓石中Nanobacteria细菌的分离、培养与鉴定

目的 从髓石中分离Nanobacteria，并进行培养和鉴定。方法 收集新鲜的27颗髓石分为9个样本，每个样本含3颗髓石，进行Nanobacteria的分离、培养，观察该菌形态学及生长特性，采用VOBKossa染色、Nanobacteria单克隆抗体8D10和G188免疫组化和间接免疫荧光染色、Hoechst染色和VOBKossa染色双重染色法联合进行Nanobacteria鉴定。结果 本次实验7个样本中所分离、培养出的细菌形态学特征与生长特性与Nanobacteria相似，VOBKossa染色呈阳性反应，单克隆抗体免疫组化染色和间接免疫荧光染色阳性。双重染色实验中，样本Hoechst染色呈阴性反应，VOBKossa染色呈阳性反应，而牙髓细胞的Hoechst染色可见胞核呈明显阳性反应，有2个样本中未见细菌生长。结论 髓石中可分离得到Nanobacteria，该菌的生物学特性可能在髓石的形成中起重要作用。

Isolation, cultivation and initial identification of Nanobacteria from dental pulp stone

Objective To isolate Nanobacteria from dental pulp stone and perform culturing and the identification of Nanobacteria. Methods Freshly collected 27 dental pulp stones were divided into nine samples. Each sample contained three dental pulp stones. All samples were used for the isolation and culture of Nanobacteria. The shape and the growth characteristics of the cultured bacteria were observed. Nanobacteria were identified by von Kossa staining, immunohistochemical staining and indirect immunofluorescence staining, double staining including Hoechst staining and von Kossa staining. Results The characteristics growth and morphology of the bacteria detected in seven samples were similar to Nanobacteria. von Kossa staining, immunohistochemical staining, indirect immunofluorescent staining were positive for Nanobacteria. In double staining method, Hoechst staining of the samples was negative for Nanobacteria, but von Kossa staining was positive. Hoechst staining of the dental pulp cells was positive. No Nanobacteria was found in the other two samples. Conclusions The bacteria isolated from dental pulp stone in this study was similar to Nanobacteria in terms of growth rate , morphology and staining properties. These unusual properties of the bacteria may play an important role in the formation of pulp stone.