Propionibacterium acnes biofilm is present in intervertebral discs of patients undergoing microdiscectomy.
Propionibacterium acnes biofilm is present in intervertebral discs of patients undergoing microdiscectomy.
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DOI：
10.1371/journal.pone.0174518
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Slaby O
中科院分区：
文献类型：
--
作者：
Capoor MN;Ruzicka F;Schmitz JE;James GA;Machackova T;Jancalek R;Smrcka M;Lipina R;Ahmed FS;Alamin TF;Anand N;Baird JC;Bhatia N;Demir-Deviren S;Eastlack RK;Fisher S;Garfin SR;Gogia JS;Gokaslan ZL;Kuo CC;Lee YP;Mavrommatis K;Michu E;Noskova H;Raz A;Sana J;Shamie AN;Stewart PS;Stonemetz JL;Wang JC;Witham TF;Coscia MF;Birkenmaier C;Fischetti VA;Slaby O
在以前的研究中，痤疮丙酸杆菌是从约25%接受显微椎间盘切除术的患者的椎间盘组织中培养出来的，这表明慢性细菌感染和椎间盘退变之间可能存在联系。然而，考虑到痤疮丙酸杆菌作为皮肤真菌的重要性，此类分析通常难以排除这些微生物代表围手术期微生物污染的替代可能性。本研究旨在验证切除椎间盘培养物中痤疮丙酸杆菌的患病率，同时提供椎间盘中痤疮丙酸杆菌生物膜的显微镜证据。将368例因椎间盘突出症接受显微椎间盘切除术的患者的标本分成几个片段，一个片段被均质化，进行定量厌氧培养，并评估细菌生长，第二个片段被冷冻用于其他分析。通过MALDI-TOF质谱鉴定菌落，并通过多重PCR进行痤疮丙酸杆菌分型。对于一个子集的标本，细菌定位在光盘进行了评估，通过显微镜使用共聚焦激光扫描和FISH。从162个椎间盘（44%）中培养出细菌，其中119例（32.3%）为痤疮丙酸杆菌。在89例病例中，痤疮丙酸杆菌是专门培养的;在30例病例中，它与其他细菌（主要是凝固酶阴性葡萄球菌）联合分离。在阳性标本中，痤疮丙酸杆菌细菌负荷中位数为350 CFU/g（12 - 20，000 CFU/g）。对38株痤疮丙酸杆菌分离株进行分子亚型分型，确定6个确定的致病菌群中的4个：IA 1、IB、IC和II。八个培养阳性标本进行了评价，荧光显微镜，并显示痤疮丙酸杆菌原位。值得注意的是，这些细菌在椎间盘基质内表现出生物膜分布。痤疮丙酸杆菌在男性中的流行率高于女性（39% vs. 23%，p = 0.0013）。本研究证实痤疮丙酸杆菌在椎间盘突出组织中普遍存在。此外，它提供了此类标本中痤疮丙酸杆菌生物膜的第一个视觉证据，与感染而不是微生物污染一致。
In previous studies, Propionibacterium acnes was cultured from intervertebral disc tissue of ~25% of patients undergoing microdiscectomy, suggesting a possible link between chronic bacterial infection and disc degeneration. However, given the prominence of P. acnes as a skin commensal, such analyses often struggled to exclude the alternate possibility that these organisms represent perioperative microbiologic contamination. This investigation seeks to validate P. acnes prevalence in resected disc cultures, while providing microscopic evidence of P. acnes biofilm in the intervertebral discs. Specimens from 368 patients undergoing microdiscectomy for disc herniation were divided into several fragments, one being homogenized, subjected to quantitative anaerobic culture, and assessed for bacterial growth, and a second fragment frozen for additional analyses. Colonies were identified by MALDI-TOF mass spectrometry and P. acnes phylotyping was conducted by multiplex PCR. For a sub-set of specimens, bacteria localization within the disc was assessed by microscopy using confocal laser scanning and FISH. Bacteria were cultured from 162 discs (44%), including 119 cases (32.3%) with P. acnes. In 89 cases, P. acnes was cultured exclusively; in 30 cases, it was isolated in combination with other bacteria (primarily coagulase-negative Staphylococcus spp.) Among positive specimens, the median P. acnes bacterial burden was 350 CFU/g (12 - ~20,000 CFU/g). Thirty-eight P. acnes isolates were subjected to molecular sub-typing, identifying 4 of 6 defined phylogroups: IA1, IB, IC, and II. Eight culture-positive specimens were evaluated by fluorescence microscopy and revealed P. acnes in situ. Notably, these bacteria demonstrated a biofilm distribution within the disc matrix. P. acnes bacteria were more prevalent in males than females (39% vs. 23%, p = 0.0013). This study confirms that P. acnes is prevalent in herniated disc tissue. Moreover, it provides the first visual evidence of P. acnes biofilms within such specimens, consistent with infection rather than microbiologic contamination.