An in vitro biofilm model of subgingival plaque
An in vitro biofilm model of subgingival plaque
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DOI：
10.1111/j.1399-302x.2007.00336.x
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发表时间：
2007-06-01
影响因子：
--
通讯作者：
Sedlacek, M. J.
中科院分区：
文献类型：
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作者：
Walker, C.;Sedlacek, M. J.
许多生物膜模型已经被描述用于研究与龈上菌斑相关的细菌。然而，用于龈下菌斑研究的模型较少。本研究的目的是开发和验证一个模型，密切模仿牙龈下菌群的组成。方法：采用10%无菌唾液包被羟基磷灰石钙片过夜，置于含有胰酶-豆浆的平底组织培养板中，直接接种少量分散的龈下菌斑，厌氧培养，每隔48 h转移到新鲜培养基中，直至形成高潮（稳态）生物膜（类似于10天）。结果：该模型基于8名牙周炎患者和8名健康受试者的样本，产生了一种多物种、异质性的生物膜，包括革兰氏阳性和革兰氏阴性物种，包括15-20种与龈下菌群相关的可培养物种。存在的物种及其比例反映了最初可培养的龈下菌群。健康受试者的初始斑块样本与成熟生物膜的比较显示，物种相似度为81%，比例相似度为70%。从牙周病患者获得的样品形成的生物膜在种类上有69%相似，在目前的比例上有57%相似。结论：本文所描述的生物膜模型能很好地再现可培养龈下菌斑的组成及其相对比例。从患病部位和非患病部位生长的生物膜之间存在差异，前者的特征是牙周病原体在微生物水平上显著存在。
Introduction: Numerous biofilm models have been described for the study of bacteria associated with the supragingival plaque. However, there are fewer models available for the study of subgingival plaque. The purpose of this study was to develop and validate a model that closely mimicked the composition of the subgingival flora.Methods: The model was developed as follows: calcium hydroxyapatite disks were coated overnight with 10% sterile saliva, placed in flat-bottomed tissue culture plates containing trypticase-soy broth, directly inoculated with a small aliquot of dispersed subgingival plaque, incubated anaerobically, and transferred to fresh medium at 48-h intervals until climax (steady-state) biofilms were formed (similar to 10 days).Results: The model, based on samples from eight periodontitis patients and eight healthy subjects, yielded a multi-species, heterogeneous biofilm, consisting of both gram-positive and gram-negative species, and comprising 15-20 cultivable species associated with the subgingival flora. The species present and their proportions were reflective of the initial cultivable subgingival flora. Comparisons of the initial plaque samples from healthy subjects and the mature biofilms showed 81% similarity in species and 70% similarity in the proportions present. Biofilms formed from samples obtained from periodontally diseased subjects were 69% similar in species and 57% similar in the proportions present.Conclusion: The biofilm model described here closely reproduces the composition of the cultivable subgingival plaque both in the species present and in their relative proportions. Differences existed between biofilms grown from diseased and non-diseased sites with the former being characterized by the presence of periodontal pathogens at microbially significant levels.