牙周病患者牙结石中纳米级矿化颗粒的分离培养及形态学观察

Separation,cultivation and morphological observation of Calcifying nanoparticles (CNPs) from periodontal patients' dental calculus in vitro

目的对不同程度牙周病患者牙结石中纳米级矿化颗粒分离培养,探讨该物质与牙周病之间的相关性。方法根据牙周炎症程度不同分为牙龈炎组、轻度牙周炎组、中重度牙周炎组,每组10例,分别采集病人下前牙舌面及上颌第一磨牙颊面龈上牙石、龈下牙石,其中牙龈炎组只采集以上牙位龈上牙石,分离培养后进行光镜、HE染色、茜素红钙染色及透射电镜观察。结果分离培养率比较:牙龈炎、轻度牙周炎、中重度牙周炎患者分离培养率分别为10%、70%、80%,差异具有统计学意义(P<0.01),组间比较中重度牙周炎组分离培养率明显高于牙龈炎组(P<0.0167);轻度牙周炎和中重度牙周炎组龈上结石培养阳性率为20.00%,龈下结石为62.50%,差异具有统计学意义(2=14.91,P<0.01);不同牙位及不同性别间分离培养率差异无统计学意义;HE染色光学显微镜下见颗粒周围呈深蓝色,茜素红钙染色为橘色圆形小颗粒;电镜显示其为周围有一层高密度影的圆形、卵圆形、短杆状颗粒。结论中重度牙周炎组牙结石中分离培养出的纳米级矿化颗粒明显高于牙龈炎组,轻度牙周炎和中重度牙周炎组龈下结石纳米级矿化颗粒分离培养率高于龈上结石,纳米级矿化颗粒与牙周炎之间的相关性值得进一步探讨。

Objective To separate and cultivate calcifying nanoparticles（ CNPs） from periodontal patients with various degrees and explore the relationship between CNPs and periodontal diseases. Methods Supragingival calculus,subgingival calculus at lingual of incisor and buccal of first maxillary molar from gingivitis group,gently periodontal disease group and severe periodontal disease group with 10 cases in each group were collected,respectively. The morphological observation of CNPs was performed with light microscope,HE and alizarin red staining and transmission electron microscope（ TEM）. Results The CNPs cultivation rate in gingivitis,gently periodontal disease and severe periodontal disease groups were about 10%,70% and 80%,respectively,with the statistically significant difference（ P〈0. 01） and there was significant difference of the cultivation rate between gingivitis and severe periodontal disease group（ P〈0. 0167）. The positive cultivation rate of supragingival calculus and subgingival calculus were about 20. 00% and 62. 50%,respectively,with the statistically significant difference（Х^2= 14. 91,P〈0. 01）. However,there was no significant difference of the CNPs cultivation rate in different tooth position and gender. Mazarine and orange were shown around CNPs through HE and alizarin red staining,respectively. High- density cicular,bicular- ovate or rod- shaped paticle around CNPs were indicated via TEM. Conclusion The positive cultivation rate of CNPs separated from severe periodontal disease is higher than that of gingivitis and the positive cultivation rate in supragingival calculus group is higer than that in subgingival calculus group,suggesting that the relationship between CNPs and periodontal diseases warrants further investigation.