Multi-species bacterial biofilm and intracellular infection in otitis media.
Multi-species bacterial biofilm and intracellular infection in otitis media.
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DOI：
10.1186/1471-2431-11-94
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发表时间：
2011-10-24
期刊：
影响因子：
2.4
通讯作者：
Richmond PC
中科院分区：
文献类型：
--
作者：
Thornton RB;Rigby PJ;Wiertsema SP;Filion P;Langlands J;Coates HL;Vijayasekaran S;Keil AD;Richmond PC
慢性中耳炎伴积液（COME）和复发性急性中耳炎（rAOM）患儿中耳积液中存在代谢活跃但无法通过抗生素治疗培养和根除的细菌。这些观察结果提示生物膜的存在或细菌在细胞内的隔离，并可能在OM的发病机制中发挥作用。本研究的目的是为中耳COME或rAOM患儿中耳黏膜细胞内或生物膜内存在耳致病菌提供证据。采用透射电子显微镜（TEM）、物种特异性荧光原位杂交（FISH）和共聚焦激光扫描显微镜（CLSM）对20例患有COME或rAOM的儿童中耳粘膜活检进行耳致病菌（生物膜或细胞内）检查。一个接受人工耳蜗手术的儿童的健康对照活检也被检查。健康对照样品未检出细菌。在3个活检中，有2个用TEM成像，在上皮细胞内含有液泡的粘液中观察到细菌。其中的细菌种类无法确定，也未观察到生物膜。使用FISH和CLSM， 17例中耳炎粘膜标本中有15例可见细菌。在该组中，17例中耳粘膜活检中有11例（65%）显示细菌生物膜，12例显示细胞内细菌。52%的活组织检查生物膜和细胞内细菌均阳性。在15个有细菌存在的样本中，有13个样本至少鉴定出一种耳病原体。COME患儿和rAOM患儿的活检结果无差异。使用FISH和CLSM，在患有COME和/或rAOM的儿童中耳黏膜上/中发现了已知耳病原体的细菌生物膜和细胞内感染。虽然它们在疾病发病机制中的作用仍有待确定，但这种先前未描述的感染模式可能有助于解释当前治疗策略在预防或解决COME或rAOM方面的无效。
Bacteria which are metabolically active yet unable to be cultured and eradicated by antibiotic treatment are present in the middle ear effusion of children with chronic otitis media with effusion (COME) and recurrent acute otitis media (rAOM). These observations are suggestive of biofilm presence or intracellular sequestration of bacteria and may play a role in OM pathogenesis. The aim of this project is to provide evidence for the presence of otopathogenic bacteria intracellularly or within biofilm in the middle ear mucosa of children with COME or rAOM. Middle ear mucosal biopsies from 20 children with COME or rAOM were examined for otopathogenic bacteria (either in biofilm or located intracellularly) using transmission electron microscopy (TEM) or species specific fluorescent in situ hybridisation (FISH) and confocal laser scanning microscopy (CLSM). One healthy control biopsy from a child undergoing cochlear implant surgery was also examined. No bacteria were observed in the healthy control sample. In 2 of the 3 biopsies imaged using TEM, bacteria were observed in mucus containing vacuoles within epithelial cells. Bacterial species within these could not be identified and biofilm was not observed. Using FISH with CLSM, bacteria were seen in 15 of the 17 otitis media mucosal specimens. In this group, 11 (65%) of the 17 middle ear mucosal biopsies showed evidence of bacterial biofilm and 12 demonstrated intracellular bacteria. 52% of biopsies were positive for both biofilm and intracellular bacteria. At least one otopathogen was identified in 13 of the 15 samples where bacteria were present. No differences were observed between biopsies from children with COME and those with rAOM. Using FISH and CLSM, bacterial biofilm and intracellular infection with known otopathogens are demonstrated on/in the middle ear mucosa of children with COME and/or rAOM. While their role in disease pathogenesis remains to be determined, this previously undescribed infection pattern may help explain the ineffectiveness of current treatment strategies at preventing or resolving COME or rAOM.