三种普雷沃菌致龋效果比较研究  

Comparative study on the cariogenic effects of three species of Prevotella

作　　者：赵晓芳 陶匀雅 孙雁斐 孙翔琨 梅昆洲 张雅萌 崔严文 杨芳[1,2] ZHAO Xiao-fang;TAO Yun-ya;SUN Yan-fei;SUN Xiang-kun;MEI Kun-zhou;ZHANG Ya-meng;CUI Yan-wen;YANG Fang(School of Stomatology,Qingdao University,Qingdao266003,China;不详)

机构地区：[1]青岛大学口腔医学院,山东青岛266003 [2]青岛大学青岛市市立医院口腔科,山东青岛266071 [3]山东第二医科大学口腔医学院,山东潍坊261053 [4]滨州医学院口腔医学院,山东滨州256600 [5]中国科学院青岛生物能源与过程研究所,山东青岛266101

出　　处：《中国实用口腔科杂志》2025年第2期160-169,共10页Chinese Journal of Practical Stomatology

基　　金：国家自然科学基金(81670979)。

摘　　要：目的 比较研究3种普雷沃菌[栖组织普雷沃菌(Prevotella histicola,P.h)、栖牙普雷沃菌(Prevotella denticola,P.d)、小斑点普雷沃菌(Prevotella maculosa,P.m)]的致龋效果,为龋病的预防和治疗措施提供参考依据。方法培养3种普雷沃菌和变异链球菌(Streptococcus mutans,S.m)24 h,测定细菌上清液pH值,并使用高效液相色谱仪检测上清液中细菌产酸情况。将4种菌液分别置于pH值为4.5、5.5和6.5条件下培养24 h,测定光密度(OD)值来评价细菌耐酸情况。使用结晶紫染色检测4种菌液培养48 h后生物膜形成量,扫描电镜和激光共聚焦显微镜下观察生物膜形态与结构,蒽酮法测定生物膜水不溶性胞外多糖含量。收集2024年7—8月于青岛大学青岛市市立医院口腔科就诊患者拔除的阻生第三磨牙20颗,制备40个牙釉质块随机分为5组,每组8个,分别置于含P.h(P.h组)、P.d(P.d组)、P.m(P.m组)、S.m(S.m组)菌液及无菌(对照组)的TSBS培养液中。扫描电镜下观察培养2周后的牙釉质表面形态,使用显微维氏硬度计测定牙釉质块培养前后的表面维氏硬度(HV)差值。结果 (1)培养24 h后,S.m相较于3种普雷沃菌,上清液pH值较小,乳酸和总产酸质量浓度较大;P.h上清液pH值显著小于P.d和P.m,琥珀酸、乳酸、乙酸及总产酸质量浓度均大于P.d和P.m,差异均有统计学意义(均P <0.05);而P.d和P.m比较,差异无统计学意义(P> 0.05)。(2)不同pH条件下培养24 h后,S.m相较于3种普雷沃菌的OD值明显增大;当pH=6.5时,P.h的OD值大于P.d和P.m,P.d的OD值大于P.m,差异均有统计学意义(均P <0.05);随着pH值的降低,细菌的生长逐渐受到抑制,当pH=4.5时,S.m的OD值增长缓慢,而3种普雷沃菌的生长被严重抑制。(3)培养48 h后,3种普雷沃菌均能形成生物膜。P.h形成的生物膜具有均匀致密的特点,由大量微生物菌落构成。与P.d和P.m相比,P.h生物膜中水不溶性胞外多糖质量浓度[(1.39±0.23)g/L]显著升高,�ObjectiveTo investigate and compare the cariogenic effects of three Prevotella species[Prevotella histicola(P.h),Prevotella denticola(P.d)and Prevotella maculosa(P.m)] and provide a reference basis for the prevention and treatment of dental caries.Methods Three Prevotella species and Streptococcus mutans(S.m)were cultured for 24 h.The pH of the bacterial supernatant was then measured,and high-performance liquid chromatography was employed to detect acid production in the supernatant.The four bacterial cultures were incubated at pH levels of 4.5,5.5,and 6.5 for 24 h,and the optical density(OD)was measured to evaluate bacterial acid resistance.Crystal violet staining was used to detect the biofilm formation of the four bacterial cultures after 48 h of incubation.The morphology and structure of the biofilm were observed using scanning electron microscopy and laser confocal microscopy.The content of water-insoluble extracellular polysaccharides in the biofilm was determined using the anthrone method.Twenty impacted third molars were collected from patients who visited the Department of Stomatology at Qingdao Municipal Hospital between July and August 2024.Forty enamel blocks were prepared and randomly divided into five groups,each containing eight blocks.The groups were incubated in TSBS culture medium with bacterial solutions of P.h(P.h group),P.d(P.d group),P.m(P.m group),and S.m(S.m group),and a sterile control group was set.After two weeks of culture,the enamel surface morphology was examined using scanning electron microscope,and the difference in Vickers hardness(HV)of the enamel blocks before and after culturing was measured with a micro Vickers hardness tester.Results After 24 h of incubation,compared with the three Prevotella species,the pH of the supernatant from S.m was lower,and the concentrations of lactic acid and total acid were higher.The pH of the supernatant from P.h was significantly lower than that from P.d and P.m,and the concentrations of succinic acid,lactic acid,acetic acid,and total acid were

关 键 词：栖组织普雷沃菌 栖牙普雷沃菌 小斑点普雷沃菌 牙釉质龋 生物膜 脱矿 

分 类 号：R78[医药卫生—口腔医学]