Hertel Susann, Wolf Alexandra, Basche Sabine, Viergutz Gabriele, Rupf Stefan, Hannig Matthias, Hannig Christian
Clinic of Operative and Pediatric Dentistry Medical Faculty Carl Gustav Carus, Technical University Dresden, Germany.
Dental Practice, Berlin, Germany.
Am J Dent. 2017 Jun;30(3):171-176.
To investigate patterns of overnight in situ microbial colonization of enamel in children.
Overall, 29 children (aged 5-9 years) participated in the study. Nine were caries-free with no decayed, missing, or filled teeth (DMFT), 11 were caries-rehabilitated (DMFT ≥ 2, no active carious lesions), and nine were caries-active (DMFT ≥ 2, at least two carious lesions). Bovine enamel samples were fixed on individual upper jaw splints stored overnight in situ. 4',6-diamidino-2-phenylindole (DAPI) combined with Concanavalin A staining was applied for fluorescence microscopic visualization of total adherent bacteria and glucans. Fluorescence in situ hybridization (FISH) was used for distinction of eubacteria, streptococci, and Candida albicans. Salivary samples were investigated for Streptococcus mutans (S. mutans) by using CRT bacteria test and yeasts with Calcofluor white (CFW) staining.
With all fluorescence methods, bacteria but not Candida albicans were detected on enamel samples. No statistically significant differences were observed in distribution patterns of the adherent bacteria between the groups. CFW staining indicated fungal structures in saliva samples of all participants. Based on CRT test results, the lowest amount of S. mutans were observed in caries-free children. Thus, initial microbial colonization patterns of enamel in children are not influenced by caries activity.
Caries activity in children may influence the process of initial bioadhesion and thus distribution patterns of bacterial attachment to the enamel surface. Investigation of in situ biofilm formation might provide valuable insights regarding the varying caries susceptibility in children.
研究儿童牙釉质过夜原位微生物定植模式。
共有29名儿童(5 - 9岁)参与本研究。其中9名无龋,无龋坏、缺失或充填牙(DMFT);11名已进行龋病修复(DMFT≥2,无活动性龋损);9名患有活动性龋病(DMFT≥2,至少有两个龋损)。将牛牙釉质样本固定在个体上颌夹板上,原位储存过夜。采用4',6-二脒基-2-苯基吲哚(DAPI)与伴刀豆球蛋白A染色相结合的方法,通过荧光显微镜观察总黏附细菌和葡聚糖。采用荧光原位杂交(FISH)技术区分真细菌、链球菌和白色念珠菌。使用CRT细菌检测法检测唾液样本中的变形链球菌(S. mutans),并用荧光增白剂(CFW)染色检测酵母菌。
采用所有荧光方法,在牙釉质样本上均检测到细菌,但未检测到白色念珠菌。各组之间黏附细菌的分布模式未观察到统计学显著差异。CFW染色显示所有参与者唾液样本中有真菌结构。根据CRT检测结果,无龋儿童中变形链球菌的数量最少。因此,儿童牙釉质的初始微生物定植模式不受龋病活动的影响。
儿童的龋病活动可能影响初始生物黏附过程,进而影响细菌在牙釉质表面附着的分布模式。对原位生物膜形成的研究可能为了解儿童不同的龋病易感性提供有价值的见解。