A Novel Organotypic Dento-Epithelial Culture Model: Effect of Fusobacterium nucleatum Biofilm on B-Defensin-2,-3, and LL-37 Expression
A Novel Organotypic Dento-Epithelial Culture Model: Effect of Fusobacterium nucleatum Biofilm on B-Defensin-2,-3, and LL-37 Expression
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DOI：
10.1902/jop.2011.110177
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发表时间：
2012-02-01
影响因子：
4.3
通讯作者：
Uitto, Veli-Jukka
中科院分区：
文献类型：
--
作者：
Gursoy, Ulvi K.;Pollanen, Marja;Uitto, Veli-Jukka
背景资料：在本研究中，三个上皮肽（人β-防御素[hBD]-2和-3，cathelicidin [LL-37]）的表达和定位研究在器官型牙上皮（OD-E）模型暴露于具核梭杆菌（Fn）biofilm.Methods：生物膜的Fn ATCC 25586或AHN 9508通过培养每个菌株在半透膜。OD-E模型通过将角质形成细胞接种在含有成纤维细胞的胶原凝胶上并将牙本质块放置在顶部来构建。将3天大的生物膜置于OD-E的顶部，并将共培养物孵育5小时或24小时。结果：培养5小时后，各Fn菌株的生物膜刺激hBD-2和hBD-3的表达。hBD-2定位于上皮和牙本质上皮连接的表层，hBD-3定位于基底细胞层，而LL-37仅弱表达。24小时后，hBD-2表达向上皮的基底细胞层延伸。相反，hBD-3的表达延伸到上皮的浅层。在Fn AHN 9508生物膜的情况下，LL-37被定位在dento-epithelial junction.Conclusion的细胞层中：在我们的OD-E模型中，上皮抗菌肽对Fn生物膜的反应具有独特的调节和定位特征，类似于已知的在体内牙龈上皮中发生的那些。牙周病学杂志2012;83：242-247。
Background: In the present study, the expression and localization of three epithelial peptides (human p-defensin [hBD]-2 and -3, and cathelicidin [LL-37]) are studied in an organotypic dento-epithelial (OD-E) model exposed to Fusobacterium nucleatum (Fn) biofilm.Methods: Biofilm of Fn ATCC 25586 or AHN 9508 were produced by culturing each strain on semipermeable membranes. The OD-E model was constructed by seeding keratinocytes on fibroblast-containing collagen gels and by placing dentin pieces on the top. A 3-day-old biofilm was placed on the top of the OD-E and the coculture was incubated for 5 hours or 24 hours. Production of epithelial antimicrobial peptides was determined immunohistochemically.Results: After 5 hours of incubation, the biofilm of each Fn strain stimulated expression of hBD-2 and -3. hBD-2 was localized on superficial layers and hBD-3 on basal cell layers of the epithelium and dento-epithelial junctions, whereas LL-37 was only weakly expressed. After 24 hours, hBD-2 expression was extended toward basal cell layers of the epithelium. In contrast, hBD-3 expression extended toward superficial layers of the epithelium. In the case of Fn AHN 9508 biofilm, LL-37 was localized in the cell layers of the dento-epithelial junction.Conclusion: In our OD-E model, epithelial antimicrobial peptide responses to Fn biofilms have distinct regulation and localization characteristics, resembling those known to occur in the gingival epithelium in vivo. J Periodontot 2012;83:242-247.