SD大鼠胫骨慢性骨髓炎动物模型的构建及效果评价

发布时间：2018-01-03 01:39

本文关键词：SD大鼠胫骨慢性骨髓炎动物模型的构建及效果评价　出处：《广州中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:用简易的方法构建一个稳定可复制的SD大鼠胫骨慢性骨髓炎动物模型。方法:30只SD大鼠随机分为实验组(n=20)与对照组(n=10)两组。3%戊巴比妥钠溶液腹腔注射麻醉后,右后腿备皮,碘酒消毒,铺无菌巾,胫骨近侧干骺端内下方切0.5cm纵行切口,显露内侧骨皮质,在胫骨结节内下0.5cm处钻一小孔。往实验组大鼠右胫骨骨髓腔内注入0.1ml5%鱼肝油酸钠和0.1ml2× 108/ml菌落形成单位(CFU)的标准金黄色葡萄球菌(ATCC25934)悬液;往对照组(n=10)大鼠的右胫骨髓腔内注入0.1ml5%鱼肝油酸钠和0.1ml生理盐水。观察术后一周和三周大鼠体重、病灶大体感染情况、造模胫骨X线表现;对所拍X线片由放射科医师独立进行Norden评分。术后第一周从实验组和对照组分别随机抽取5只SD大鼠,过量麻醉处死后,取造模灶周围软组织做细菌培养,取右胫骨标本做组织病理学切片HE染色分析。术后第三周,过量麻醉处死所有大鼠,取造模灶周围软组织做细菌培养,取右胫骨标本做组织病理学切片HE染色分析。用IBM SPSS 19版本软件进行统计学分析。多次重复测量采用重复测定方差分析的统计学方法;术后第一周、术后第三周实验组与对照组右胫骨X线Norden评分比较用非参数检验两独立样本Mann-Whitney U检验法的统计学方法。实验组和对照组术后第一周与术后第三周X线Norden评分比较用非参数检验Wilcomxon配对符秩检验。结果:术后第一周:实验组大鼠体重比对照组大鼠体重轻,差异有统计学意义。实验组所有动物在手术区域大体感染情况明显,手术切口未愈合,对照组手术切口基本愈合。实验组动物右胫骨的X线摄片可见明显的骨髓内高亮度影,有些可见骨膜反应,未见明显的骨缺损和死骨形成,Norden评分平均分1.15±0.587;对照组大鼠右胫骨X线片结果与正常侧胫骨X线表现基本无异,Norden评分平均分为0.30±0.483;实验组大鼠右胫骨X线片Norden评分明显高于对照组大鼠右胫骨X线Norden评分,差异具有统计学意义。随机抽取实验组5只大鼠的胫骨标本组织病理学切片HE染色光镜下可见视野中大量炎症细胞浸润、骨膜增生;对照组的胫骨标本病理切片视野中偶可见炎症细胞。实验组5只大鼠的造模病灶周围软组织标本细菌培养全为阳性;对照组全为阴性。术后第三周:实验组大鼠体重与对照组大鼠无明显差异。实验组所有动物在手术区域大体感染情况明显趋于慢性,4只大鼠病灶出现窦道,其余皮肤手术切口基本愈合,但大部分呈现明显色素沉着;对照组动物手术切口完全愈合,手术区域与正常组织基本无异。实验组大鼠右胫骨X线片大多数可见严重的骨膜反应、新骨形成、骨质破坏和死骨形成,有些可见骨干畸形,Norden评分平均分为2.67±0.617;对照组X线片与正常侧无异,Norden评分平均分为0;实验组大鼠右胫骨X线片Norden评分明显高于对照组大鼠右胫骨X线Norden评分,差异具有统计学意义。实验组右胫骨标本组织病理学切片HE染色光镜下可见大量炎症细胞浸润、严重骨细胞破坏、骨膜增生;对照组骨组织病理学切片可见排列紧密、有秩序的骨细胞排列。实验组所有大鼠的造模病灶周围软组织标本细菌培养全为阳性,对照组全为阴性。术后第三周实验组大鼠的右胫骨X线Norden评分明显高于第一周实验组大鼠的右胫骨X线Norden评分,差异有统计学意义。结论:该造模方法稳定可行,可在往SD大鼠胫骨近侧干骺端髓腔内注入0.1ml5%鱼肝油酸钠和0.1ml2× 108CFU/mL金黄色葡萄球菌后的第三周时成功造出SD大鼠胫骨慢性骨髓炎动物模型。
[Abstract]:Objective: to construct SD rat animal model of chronic osteomyelitis of tibia can copy a stable using a simple method. Methods: 30 SD rats were randomly divided into experimental group (n=20) and control group (n=10) anesthesia group two.3% after intraperitoneal injection of pentobarbital sodium, right hind leg skin preparation, iodine disinfection, shop the aseptic towel, the proximal tibial metaphysis below the cut 0.5cm longitudinal incision, expose the medial cortical bone, the tibial tubercle in 0.5cm drill a small hole. To the experimental rats right tibia bone marrow cavity injection of 0.1ml5% Cod Liver Oil acid sodium and 0.1ml2 * 108/ml colony forming units (CFU) standard Staphylococcus aureus (ATCC25934) suspension; to control group (n=10) rats right tibia bone marrow cavity injection of 0.1ml5% Cod Liver Oil acid sodium and 0.1ml saline. The body weight and three week rats were observed after lesions generally infection model of tibia X-ray; to shoot X ray independently by a radiologist Norden score. After the first week of the experimental group and control group were randomly selected 5 SD rats were killed by overdose anesthesia after the molding around the focus of soft tissue for bacterial culture, take the right tibia specimens of pathological HE staining analysis. After third weeks, all rats were killed by overdose anesthesia. The modeling of soft tissue around the focus of bacterial culture, take the right tibia specimens for histological analysis. HE staining was analyzed by IBM SPSS software version 19. Repeated measurements analysis of variance by statistical method of repeated determination; after the first week, third weeks after operation, the experimental group and the control group the right tibia X-ray Norden score comparison test test of two independent samples Mann-Whitney U using non parametric statistical methods. The experimental group and the control group after the first week and third weeks after operation, X-ray Norden scores were compared by nonparametric test Wilcomxon paired rank test. Test. Results: after the first week: the body weight of rats in the experimental group than the control group rats weight, the difference was statistically significant. The experimental group in the operation area in all animal infection, incision healing, incision healed in control group. The experimental group animal right tibia X-ray showed bone marrow in the shadow of high brightness, some visible periosteal reaction, there was no obvious bone defect and dead bone formation, the average Norden score was 1.15 + 0.587; the control group rats right tibia X-ray results and normal tibia X-ray manifestations of basically the same, the average Norden score was divided into 0.30 + 0.483; the experimental group rats right tibia X-ray Norden the score was significantly higher than the control group rats right tibia X-ray Norden score, the difference was statistically significant. The experimental group were randomly selected from the pathological specimens of tibia in 5 rats, HE staining of inflammatory cells under light microscope in the field of leaching Run, periosteal proliferation; even visible inflammatory cells in the control group tibia biopsy specimens. The experimental group in the field of die lesions surrounding soft tissue samples of bacterial culture was made all of the 5 rats in the control group; all negative. Third weeks after operation: the weight of the experimental group rats and control group rats had no significant difference all animal experimental group. In the operation area in infection tends to be chronic, 4 rats were seen in sinus, the remaining skin incision healed, but the most obvious pigmentation; the control group animal surgical incision healed completely, the operation area is basic and normal tissues. The experimental group rats right tibia X-ray most visible severe periosteal reaction and new bone formation, bone destruction and dead bone formation, some visible backbone deformity, the average Norden score was divided into 2.67 + 0.617; the control group is normal and lateral X-ray, Norden score score is 0;. The experimental group rats right tibia X-ray Norden score was significantly higher than that of the control group rats right tibia X-ray Norden score, the difference was statistically significant. The experimental group right tibia specimens histopathological sections of HE staining under light microscope, inflammatory cell infiltration, severe bone cell destruction, periosteal proliferation; the control sections visible closely arranged group of bone tissue the pathology of bone cells arranged in order of the experimental group. All rats model of soft tissue around the lesion samples of bacterial culture was positive, the control group was negative. After third weeks the experimental group rats right tibia X-ray Norden score was significantly higher than that in the first week of the experimental group rats right tibia X-ray Norden score, there are significant difference. Conclusion: the method is stable and feasible, can be in to the SD rat proximal tibial metaphysis intramedullary injection of 0.1ml5% Cod Liver Oil acid sodium and 0.1ml2 * 108CFU/mL of Staphylococcus aureus after third weeks The animal model of chronic osteomyelitis of tibia in SD rats was successfully made.

【学位授予单位】：广州中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R681.2;R-332

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