| 引用本文: |
吴婧艺, 李扬, 钱亚男, 张鑫源, 曹玉净.耐甲氧西林金黄色葡萄球菌感染慢性骨髓炎大鼠胫骨骨缺损模型的构建与评估[J].湖南中医药大学学报,2025,45(10):1831-1838[点击复制] |
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| 耐甲氧西林金黄色葡萄球菌感染慢性骨髓炎大鼠胫骨骨缺损模型的构建与评估 |
| 吴婧艺,李扬,钱亚男,张鑫源,曹玉净 |
| (河南中医药大学骨伤学院, 河南 郑州 450046;河南省中医院(河南中医药大学第二附属医院), 河南 郑州 450002) |
| 摘要: |
| 目的 构建评估耐甲氧西林金黄色葡萄球菌（MRSA）（ATCC 43300）感染的胫骨骨缺损大鼠慢性骨髓炎模型。方法 50只Wistar大鼠随机分为5组：对照组与MRSA感染浓度梯度组（A组1×105 CFU/mL、B组1×107 CFU/mL、C组1×109 CFU/mL、D组1×1011 CFU/mL），每组10只。在大鼠左侧胫骨制造骨缺损后采用直接注射法造模。术后4周，观察大体情况并进行Petty外观评分，ELISA检测血清C反应蛋白（CRP）含量，行X线、微计算机断层扫描（micro-CT）影像学检查并进行X线Norden骨髓炎评分，HE染色观察组织变化并进行Smeltzer病理学评分，细菌培养检测菌落计数。结果 在术后4周，与对照组相比，C组与D组的Petty外观评分均升高（P＜0.01）；与A组相比，D组Petty外观评分升高（P＜0.05）。术后4周，与对照组及A组相比，B、C与D组CRP含量均升高（P＜0.05）；与B组相比，C组和D组CRP含量均升高（P＜0.05）；与C组相比，D组CRP含量升高（P＜0.05）。术后4周，与对照组相比，C组和D组的X线Norden骨髓炎评分均升高（P＜0.01），菌落数量均增加（P＜0.01），B、C与D组Smeltzer病理学评分均升高（P＜0.01）；与A组相比，D组X线Norden骨髓炎评分升高（P＜0.01），菌落数量增加（P＜0.05），B、C和D组Smeltzer病理学评分均升高（P＜0.01）；与B组相比，C组和D组Smeltzer病理学评分均升高（P＜0.05，P＜0.01）。结论 直接注射10 μL浓度为1×109 CFU/mL的MRSA（ATCC 43300）菌液于大鼠胫骨骨缺损处，可显著升高CRP含量与细菌负荷，诱发与慢性骨髓炎相符的病理表现，成功构建出稳定可靠的慢性骨髓炎动物模型。 |
| 关键词: 耐甲氧西林金黄色葡萄球菌 慢性骨髓炎 动物模型 骨缺损 胫骨感染 模型构建 |
| DOI：10.3969/j.issn.1674-070X.2025.10.005 |
| 投稿时间：2025-08-01 |
| 基金项目:河南省科技攻关项目（252102311254，242102310108）；河南省中医药科学研究专项课题（2023ZY1008）；河南中医药大学科研创新项目（2024KYCX07）。 |
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| Establishment and evaluation of a chronic osteomyelitis model with a tibial bone defect induced by methicillin-resistant Staphylococcus aureus in rat |
| WU Jingyi, LI Yang, QIAN Ya'nan, ZHANG Xinyuan, CAO Yujing |
| (School of Orthopedics and Traumatology, Henan University of Chinese Medicine, Zhengzhou, Henan 450046, China;Henan Hospital of Traditional Chinese Medicine (The Second Hospital of Henan University of Chinese Medicine), Zhengzhou, Henan 450002, China) |
| Abstract: |
| Objective To establish and evaluate a rat model of chronic osteomyelitis induced by methicillin-resistant Staphylococcus aureus (MRSA) (ATCC 43300) infection in a tibial bone defect. Methods Fifty Wistar rats were randomized into five groups (n=10 per group): a control group and four MRSA infection groups with concentration gradients (Group A: 1×105 CFU/mL, Group B: 1×107CFU/mL, Group C: 1×109CFU/mL, Group D: 1×1011 CFU/mL). A bone defect was created in the left tibia of all rats, and the model was established using the direct injection method. At 4 weeks post-surgery, the rat general condition was observed and the wounds were assessed using the Petty appearance scoring system; serum C-reactive protein (CRP) level was measured by ELISA; X-ray and micro-computed tomography, micro-CT imaging were performed, followed by X-ray Norden osteomyelitis scoring; histopathological changes were observed via HE staining and evaluated using the Smeltzer pathological scoring system; bacterial colony counts were determined through bacterial culture. Results At 4 weeks post-surgery, compared with the control group, the Petty appearance scores of Groups C and D significantly increased (P＜0.01); compared with Group A, the Petty appearance score of Group D significantly increased (P＜0.05). At 4 weeks post-surgery, compared with the control group and Group A, the CRP levels in Groups B, C, and D were significantly elevated (P＜0.05); compared with Group B, the CRP levels in Groups C and D significantly increased (P＜0.05); compared with Group C, the CRP level in Group D was significantly higher (P＜0.05). At 4 weeks post-surgery, compared with the control group, the X-ray Norden osteomyelitis scores and bacterial colony counts in Groups C and D significantly increased (P＜0.01), and the Smeltzer pathological scores in Groups B, C, and D were significantly elevated (P＜0.01). Compared with Group A, the X-ray Norden osteomyelitis score and bacterial colony count in Group D were significantly higher (P＜0.01 and P＜0.05, respectively), and the Smeltzer pathological scores in Groups B, C, and D significantly increased (P＜0.01). Compared with Group B, the Smeltzer pathological scores in Groups C and D were significantly elevated (P＜0.05 and P＜0.01, respectively). Conclusion Direct injection of 10 μL of 1×109CFU/mL MRSA (ATCC 43300) into the rat tibial bone defect can significantly elevate the CRP level and bacterial load, inducing pathological manifestations consistent with chronic osteomyelitis, and thus can successfully establish a stable and reliable animal model for chronic osteomyelitis. |
| Key words: methicillin-resistant Staphylococcus aureus chronic osteomyelitis animal model bone defect tibial infection model establishment |
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