本文關(guān)鍵詞：SD大鼠脛骨慢性骨髓炎動物模型的構(gòu)建及效果評價　出處：《廣州中醫(yī)藥大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 慢性骨髓炎 動物模型SD大鼠 金黃色葡萄球菌

【摘要】：目的:用簡易的方法構(gòu)建一個穩(wěn)定可復(fù)制的SD大鼠脛骨慢性骨髓炎動物模型。方法:30只SD大鼠隨機(jī)分為實驗組(n=20)與對照組(n=10)兩組。3%戊巴比妥鈉溶液腹腔注射麻醉后,右后腿備皮,碘酒消毒,鋪無菌巾,脛骨近側(cè)干骺端內(nèi)下方切0.5cm縱行切口,顯露內(nèi)側(cè)骨皮質(zhì),在脛骨結(jié)節(jié)內(nèi)下0.5cm處鉆一小孔。往實驗組大鼠右脛骨骨髓腔內(nèi)注入0.1ml5%魚肝油酸鈉和0.1ml2× 108/ml菌落形成單位(CFU)的標(biāo)準(zhǔn)金黃色葡萄球菌(ATCC25934)懸液;往對照組(n=10)大鼠的右脛骨髓腔內(nèi)注入0.1ml5%魚肝油酸鈉和0.1ml生理鹽水。觀察術(shù)后一周和三周大鼠體重、病灶大體感染情況、造模脛骨X線表現(xiàn);對所拍X線片由放射科醫(yī)師獨立進(jìn)行Norden評分。術(shù)后第一周從實驗組和對照組分別隨機(jī)抽取5只SD大鼠,過量麻醉處死后,取造模灶周圍軟組織做細(xì)菌培養(yǎng),取右脛骨標(biāo)本做組織病理學(xué)切片HE染色分析。術(shù)后第三周,過量麻醉處死所有大鼠,取造模灶周圍軟組織做細(xì)菌培養(yǎng),取右脛骨標(biāo)本做組織病理學(xué)切片HE染色分析。用IBM SPSS 19版本軟件進(jìn)行統(tǒng)計學(xué)分析。多次重復(fù)測量采用重復(fù)測定方差分析的統(tǒng)計學(xué)方法;術(shù)后第一周、術(shù)后第三周實驗組與對照組右脛骨X線Norden評分比較用非參數(shù)檢驗兩獨立樣本Mann-Whitney U檢驗法的統(tǒng)計學(xué)方法。實驗組和對照組術(shù)后第一周與術(shù)后第三周X線Norden評分比較用非參數(shù)檢驗Wilcomxon配對符秩檢驗。結(jié)果:術(shù)后第一周:實驗組大鼠體重比對照組大鼠體重輕,差異有統(tǒng)計學(xué)意義。實驗組所有動物在手術(shù)區(qū)域大體感染情況明顯,手術(shù)切口未愈合,對照組手術(shù)切口基本愈合。實驗組動物右脛骨的X線攝片可見明顯的骨髓內(nèi)高亮度影,有些可見骨膜反應(yīng),未見明顯的骨缺損和死骨形成,Norden評分平均分1.15±0.587;對照組大鼠右脛骨X線片結(jié)果與正常側(cè)脛骨X線表現(xiàn)基本無異,Norden評分平均分為0.30±0.483;實驗組大鼠右脛骨X線片Norden評分明顯高于對照組大鼠右脛骨X線Norden評分,差異具有統(tǒng)計學(xué)意義。隨機(jī)抽取實驗組5只大鼠的脛骨標(biāo)本組織病理學(xué)切片HE染色光鏡下可見視野中大量炎癥細(xì)胞浸潤、骨膜增生;對照組的脛骨標(biāo)本病理切片視野中偶可見炎癥細(xì)胞。實驗組5只大鼠的造模病灶周圍軟組織標(biāo)本細(xì)菌培養(yǎng)全為陽性;對照組全為陰性。術(shù)后第三周:實驗組大鼠體重與對照組大鼠無明顯差異。實驗組所有動物在手術(shù)區(qū)域大體感染情況明顯趨于慢性,4只大鼠病灶出現(xiàn)竇道,其余皮膚手術(shù)切口基本愈合,但大部分呈現(xiàn)明顯色素沉著;對照組動物手術(shù)切口完全愈合,手術(shù)區(qū)域與正常組織基本無異。實驗組大鼠右脛骨X線片大多數(shù)可見嚴(yán)重的骨膜反應(yīng)、新骨形成、骨質(zhì)破壞和死骨形成,有些可見骨干畸形,Norden評分平均分為2.67±0.617;對照組X線片與正常側(cè)無異,Norden評分平均分為0;實驗組大鼠右脛骨X線片Norden評分明顯高于對照組大鼠右脛骨X線Norden評分,差異具有統(tǒng)計學(xué)意義。實驗組右脛骨標(biāo)本組織病理學(xué)切片HE染色光鏡下可見大量炎癥細(xì)胞浸潤、嚴(yán)重骨細(xì)胞破壞、骨膜增生;對照組骨組織病理學(xué)切片可見排列緊密、有秩序的骨細(xì)胞排列。實驗組所有大鼠的造模病灶周圍軟組織標(biāo)本細(xì)菌培養(yǎng)全為陽性,對照組全為陰性。術(shù)后第三周實驗組大鼠的右脛骨X線Norden評分明顯高于第一周實驗組大鼠的右脛骨X線Norden評分,差異有統(tǒng)計學(xué)意義。結(jié)論:該造模方法穩(wěn)定可行,可在往SD大鼠脛骨近側(cè)干骺端髓腔內(nèi)注入0.1ml5%魚肝油酸鈉和0.1ml2× 108CFU/mL金黃色葡萄球菌后的第三周時成功造出SD大鼠脛骨慢性骨髓炎動物模型。
[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.

【學(xué)位授予單位】：廣州中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R681.2;R-332

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