开发体外模型,研究金黄色葡萄球菌入侵骨细胞裂隙-髓鞘网络的情况。

IF 2.1 3区 医学 Q2 ORTHOPEDICS
Niels Vanvelk, Karen L de Mesy Bentley, Michael H J Verhofstad, Willem-Jan Metsemakers, Thomas F Moriarty, Claudia Siverino
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引用次数: 0

摘要

金黄色葡萄球菌有多种逃避宿主免疫系统和抗生素治疗的机制。其中一种机制是侵入骨细胞裂隙-颅骨网(OLCN),这可能是清创和抗生素治疗后感染复发的重要原因。本研究的目的是建立一个体外模型,以便于进一步研究金黄色葡萄球菌入侵 OLCN 的情况,并对针对这一龛位细菌的抗菌策略进行早期测试。通过显微镜测量未感染的人类、绵羊和小鼠骨骼的管腔直径,结果显示管腔直径有很大的重叠。布朗和布伦染色切片中的骨细胞裂隙以及扫描电子显微镜显示,金黄色葡萄球菌在体外成功侵入了所有物种的OLCN。然后选择小鼠骨骼进行进一步实验,将带有野生型或ΔPBP4突变型金黄色葡萄球菌USA300的钛针经皮质放置在胰蛋白酶大豆肉汤中培养2周。野生型金黄色葡萄球菌很容易侵入小鼠骨骼中的骨细胞裂隙,而ΔPBP4对OLCN的侵入明显较低(p = 0.0005)。然后用庆大霉素、西他沙星、R14噬菌体处理或不处理骨标本。庆大霉素(p = 0.0027)和西他沙星(p = 0.0280)显著降低了金黄色葡萄球菌占据裂隙的比例,而噬菌体处理则没有影响。这项研究表明,金黄色葡萄球菌能够在体外模型中侵入 OLCN。这种体外模型可用于未来的早期研究,然后再进行体内研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development of an ex vivo model to study Staphylococcus aureus invasion of the osteocyte lacuno-canalicular network.

Staphylococcus aureus has multiple mechanisms to evade the host's immune system and antibiotic treatment. One such mechanism is the invasion of the osteocyte lacuno-canalicular network (OLCN), which may be particularly important in recurrence of infection after debridement and antibiotic therapy. The aim of this study was to develop an ex vivo model to facilitate further study of S. aureus invasion of the OLCN and early-stage testing of antibacterial strategies against bacteria in this niche. The diameter of the canaliculi of non-infected human, sheep, and mouse bones was measured microscopically on Schmorl's picrothionin stained sections, showing a large overlap in canalicular diameter. S. aureus successfully invaded the OLCN in all species in vitro as revealed by presence in osteocyte lacunae in Brown and Brenn-stained sections and by scanning electron microscopy. Murine bones were then selected for further experiments, and titanium pins with either a wild-type or ΔPBP4 mutant S. aureus USA300 were placed trans-cortically and incubated for 2 weeks in tryptic soy broth. Wild-type S. aureus readily invaded the osteocyte lacunae in mouse bones while the ΔPBP4 showed a significantly lower invasion of the OLCN (p = 0.0005). Bone specimens were then treated with gentamicin, sitafloxacin, R14 bacteriophages, or left untreated. Gentamicin (p = 0.0027) and sitafloxacin (p = 0.0280) significantly reduced the proportion of S. aureus-occupied lacunae, whilst bacteriophage treatment had no effect. This study shows that S. aureus is able to invade the OLCN in an ex vivo model. This ex vivo model can be used for future early-stage studies before proceeding to in vivo studies.

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来源期刊
Journal of Orthopaedic Research®
Journal of Orthopaedic Research® 医学-整形外科
CiteScore
6.10
自引率
3.60%
发文量
261
审稿时长
3-6 weeks
期刊介绍: The Journal of Orthopaedic Research is the forum for the rapid publication of high quality reports of new information on the full spectrum of orthopaedic research, including life sciences, engineering, translational, and clinical studies.
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