冷光标记金黄色葡萄球菌胫骨感染动物模型的建立

Establishment the Luminescence staphylococcus aureus tibia infection animal model

目的建立一种简便、稳定、便于实验研究的小鼠胫骨内植物术后金黄色葡萄球菌感染动物模型。方法将20只小鼠分为A、B两组:A组为非感染组,麻醉后在右侧胫骨上1/3段矢状面中间点横向穿双侧骨皮质置入无菌钛针;B组为感染组,以相同手术方法置入冷光标记的金黄色葡萄球菌感染的钛针。两组术后进行小鼠肉眼观察评分;放射学评价,包括X线,第1、3、5、7、10、14天作IVIS检查、标本MircoCT检查;细菌学评价:术后14d处死小鼠,取出内置物作细菌菌落计数(CFU);组织学评价:包括组织学评分,病理切片作ABH、TRAP和Brownbrenn染色。结果两组术后肉眼观察评分B组明显高于A组(P<0.05)。放射学检查:两组术后X线检查内植物位置良好,A组术后IVIS检查无菌生长,B组可见局部金黄色葡萄球菌聚集,高峰从第3天持续到第7天,后逐渐回落;Mirco-CT结果 B组针道面积大于A组,周围明显有骨溶解和骨吸收。细菌学检查:术后14d内置物细菌菌落计数A组无菌生长,B组内置物培养出大量金黄色葡萄球菌。组织学检查:两组标本组织学评分B组明显高于A组(P<0.05)。病理切片通过ABH、TRAP、Brown-brenn染色进一步证实了B组感染金黄色葡萄球菌,并在局部形成骨质破坏。结论成功建立了小鼠胫骨金黄色葡萄球菌感染模型;操作简单,观察期短,结果稳定,适用于需要骨感染模型的相关实验室研究。

Objective To establish the model was a simple,stable,easy to experiment research tibial staphylococ cus aureus infection animal of mice. Methods 20 only 7 weeks Balk/c female mice were divided into A and B groups and there were 10 mice in each group. After anesthesia in group A in the right tibia lateral sagittal middle 1/3 point on wear titanium bilateral bone cortex in sterile needle,group B in the same operation method in the Luminescence staphylococcus aureus infection titanium needle. Two groups were studied after macroscopic observation score. Ra- diological evaluation,including X-ray, 1,3,5,7,10, and 14 days, the specimen Mirco-CT, IVIS check. Evaluation of bacteriology, 14 days after death mice, took implants for bacterial colony count （CFU）. Histologic evaluation： inclu- ding the histologic grading, pathological ABH, TRAP and Brown- brenn dyeing. Results The postoperative macro scopic observation,group B was significantly higher than that of group A and there were statistical significantly differences （P〈0.05）. Radiological examination that two groups of postoperative good plant location in x-rays, post- operative IVIS examination group A sterile growth,B group of visible local staphylococcus aureus, peak lasts from day 3 to 7 days,after gradually retreated, Mirco-CT results needle way bigger than group A, group B there was an obvious bone dissolution and the surrounding bone resorption. Bacteriology examination, after 14 days implants bac- terial colony counting group A sterile growth, group B implants to cultivate A large number of staphylococcus au- reus. Histological examination that group B was significantly higher than that of group A,and there were statistical significant differences （P〈 0.05）. Pathological slices through the ABH, TRAP, Brown, brenn was dyeing group B infection of staphylococcus aureus, and in the formation of local bone destruction. Conclusion The success of the mod- el of the tibial staph infections in mice model was established, the operation is simple, short observation period, the stability,is suitable for the need of bone infection model related laboratory studies.