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首页> 外文期刊>Journal of orthopaedic research >Quantitative mouse model of implant-associated osteomyelitis and the kinetics of microbial growth, osteolysis, and humoral immunity.
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Quantitative mouse model of implant-associated osteomyelitis and the kinetics of microbial growth, osteolysis, and humoral immunity.

机译：植入物相关性骨髓炎的定量小鼠模型以及微生物生长，溶骨和体液免疫的动力学。

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Although osteomyelitis (OM) remains a serious problem in orthopedics, progress has been limited by the absence of an in vivo model that can quantify the bacterial load, metabolic activity of the bacteria over time, immunity, and osteolysis. To overcome these obstacles, we developed a murine model of implant-associated OM in which a stainless steel pin is coated with Staphylococcus aureus and implanted transcortically through the tibial metaphysis. X-ray and micro-CT demonstrated concomitant osteolysis and reactive bone formation, which was evident by day 7. Histology confirmed all the hallmarks of implant-associated OM, namely: osteolysis, sequestrum formation, and involucrum of Gram-positive bacteria inside a biofilm within necrotic bone. Serology revealed that mice mount a protective humoral response that commences with an IgM response after 1 week, and converts to a specific IgG2b response against specific S. aureus proteins by day 11 postinfection. Real-time quantitative PCR (RTQ-PCR) for the S. aureus specific nuc gene determined that the peak bacterial load occurs 11 days postinfection. This coincidence of decreasing bacterial load with the generation of specific antibodies is suggestive of protective humoral immunity. Longitudinal in vivo bioluminescent imaging (BLI) of luxA-E transformed S. aureus (Xen29) combined with nuc RTQ-PCR demonstrated the exponential growth phase of the bacteria immediately following infection that peaks on day 4, and is followed by the biofilm growth phase at a significantly lower metabolic rate (p < 0.05). Collectively, these studies demonstrate the first quantitative model of implant-associated OM that defines the kinetics of microbial growth, osteolysis, and humoral immunity following infection.
机译：尽管骨髓炎（OM）在整形外科中仍然是一个严重的问题，但是由于缺乏可以量化细菌载量，细菌随时间的代谢活性，免疫力和溶骨作用的体内模型，进展受到了限制。为了克服这些障碍，我们开发了一种与植入物相关的OM的小鼠模型，其中不锈钢针涂有金黄色葡萄球菌，并通过胫骨干physi端经皮层植入。 X射线和显微CT证实了同时发生的骨溶解和反应性骨形成，这在第7天就很明显。组织学证实了与植入物相关的OM的所有标志，即：骨溶解，死骨形成和生物膜内革兰氏阳性细菌的总luc。坏死骨内。血清学研究表明，小鼠在1周后开始发生IgM应答，开始出现保护性体液应答，并在感染后第11天转化为针对特定金黄色葡萄球菌蛋白的特异性IgG2b应答。金黄色葡萄球菌特定nuc基因的实时定量PCR（RTQ-PCR）确定，感染后11天出现峰值细菌负荷。细菌载量的减少与特异性抗体的产生的巧合提示了保护性体液免疫。 luxA-E转化的金黄色葡萄球菌（Xen29）的纵向体内生物发光成像（BLI）与nuc RTQ-PCR结合显示，细菌感染后呈指数生长期，在第4天达到峰值，随后是生物膜生长期代谢率显着降低（p <0.05）。这些研究共同证明了植入物相关的OM的第一个定量模型，该模型定义了感染后微生物生长，溶骨和体液免疫的动力学。

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