Vitkov L, Hannig M, Krautgartner W D, Herrmann M, Fuchs K, Klappacher M, Hermann A
Department of Operative Dentistry and Periodontology, Saarland University, Homburg/Saar, Germany.
Lett Appl Microbiol. 2005;41(5):404-11. doi: 10.1111/j.1472-765X.2005.01787.x.
To develop a protocol for harvesting ex vivo samples of gingival-biofilm consortia and to investigate their basic characteristics.
Gingival epithelial cells with attached biofilm were collected from healthy subjects by taking a smear. The bacterial viability was estimated via the alteration of the membrane permeability and metabolic activity via the double/single-stranded nucleic acid ratio using a confocal laser-scanning microscope. Morphological analysis was performed by scanning and transmission electron microscopy. Additionally, microbiological estimations were made. The electron microscopy revealed fimbriae-mediated adhesion and the formation of a biofilm matrix. Most bacteria were viable and had a high metabolic activity.
The presented study offers an easy to follow approach for harvesting samples of gingival-biofilm consortia. The latter differs considerably from the supragingival plaque in viability and zonal distribution. Related to free-living and in vitro-grown biofilms, the gingiva-associated biofilm revealed an atypically high metabolic activity.
Biofilm fragments should possess the basic features of the entire gingiva-associated biofilm; which as yet cannot be simulated in vitro. Thus, samples of ex vivo gingival-biofilm consortia can be used to investigate the resistance of oral biofilms against antibiotics and biocides.
制定一种采集牙龈生物膜群落离体样本的方案，并研究其基本特征。
通过涂片从健康受试者中收集附着有生物膜的牙龈上皮细胞。使用共聚焦激光扫描显微镜，通过双链/单链核酸比率来评估细菌活力，该比率反映了膜通透性和代谢活性的变化。通过扫描电子显微镜和透射电子显微镜进行形态学分析。此外，还进行了微生物学评估。电子显微镜显示菌毛介导的黏附以及生物膜基质的形成。大多数细菌具有活力且代谢活性较高。
本研究提供了一种易于操作的采集牙龈生物膜群落样本的方法。牙龈生物膜群落在活力和区域分布上与龈上菌斑有很大差异。与自由生活和体外培养的生物膜相比，牙龈相关生物膜显示出异常高的代谢活性。
生物膜片段应具备整个牙龈相关生物膜的基本特征；目前尚无法在体外模拟。因此，离体牙龈生物膜群落样本可用于研究口腔生物膜对抗生素和杀菌剂的抗性。
