Detection Of Nanobacteria From Dental Calculus And Gingival Crevicular Fluid With Periodontal Disease

Objective:Nanobacteria can produce solid apatite forming mineral-coats and initiate pathologic calcification such as human renal stones and gallstones. People with periodontal disease often have dental calculus, which is also a kind of pathologic calcification in oral cavity. The objective of this study was to determine whether Nanobacteria are present in the gingival crevicular fluid (GCF) and dental calculus from subjects with periodontal disease, explore the role of Nanobacteria in the formation of dental calculus preliminarily, try to find out the relationship between Nanobacteria and periodontal disease and to become a hot reseach for the pathogenesis of periodontal disease.Methods:1. Separation and cultivation were performed in 30 dental calculus and gingival crevicular fluid (GCF) samples with periodontal disease, 15 cases was with gingivitis and other 15 cases was with periodontitis,30 gingival crevicular fluid (GCF) samples without periodontal disease as a control group.2. The samples of the culture were identified by calcium-alizarin red staining, transmission electron microscopy (TEM), scanning electron microscope (SEM), and chemical analysis were done by Energy Dispersive X-ray Detector (EDX)Results:1. After culturing for 2 months, white-colored precipitates could be seen attach to the culture tube bottom. the positive rates of GCF in periodontal disease and normal people were 70.00%(21/30)and 33.33%(10/30) respectively, the difference was significant (P<0.05).2. calcium-alizarin red staining displayed tiny black and red calcificnodus, appearing as clusters. the positive rates of GCF in periodontal disease and normal people were 60.00% (18/30) and 26.67%(8/30) respectively, the difference was significant (P <0.05).3.The calcium-staining positive rates of GCF and dental calculus in periodontal disease were 66.67%(20/30) vs 60.00%(18/30), which was no significant difference (P>0.05).4. SEM studies revealed 100-500 nm sized particles. They appeared coccoid in shape and gathered as a big stone. TEM images showed NB with a distinct dark capsule, with particles like apatite structures both within and surrounding them.5. EDX showed the paticles contained calcium, phosphonium, oxygen, carbon.Conclusion:1.Nanobacteria can be cultured and identified from dental calculus and GCF with periodontal disease, the composition of Nanobacteria is similar to dental calculus. 2. The positive rate of periodontal disease is higher than normal people, it should be further confirmed whether Nanobacteria contribute to the pathogenesis of periodontal disease.