本文選題：慢性骨髓炎 + 兔�。� 參考：《承德醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:慢性骨髓炎指骨組織的慢性感染,由外傷、交通傷等高能量損傷特別是開放性骨損傷未經(jīng)徹底清創(chuàng)或雖經(jīng)徹底清創(chuàng)但創(chuàng)傷及污染較重或由手術(shù)時無菌操作不嚴(yán)格所引起,是困擾骨科醫(yī)師的難題之一。慢性骨髓炎因其死骨形成、竇道、死腔及病變?nèi)庋拷M織的長期存在,治療較困難。目前對慢性骨髓炎的臨床治療措施是清創(chuàng)術(shù)的主要治療方法,髓腔灌洗和靜脈治療4-6周的敏感抗生素,在一個療程結(jié)束后通常能緩解部分癥狀,紅腫熱痛等炎癥表現(xiàn)可有明顯減輕,但患者常常伴有低燒,咽喉干燥,全身不適等癥狀,常規(guī)檢查提示白細(xì)胞仍處在較高值,在臨床上可理解成骨髓炎復(fù)發(fā)。骨髓炎反復(fù)發(fā)作的同時也伴隨著人體對抗生素嚴(yán)重耐藥的現(xiàn)象,造成的結(jié)果為必然抗生素的不斷升級,并由此帶來的不斷升級抗生素[4]以及再次升級再次耐藥的惡性循環(huán)。該研究實(shí)驗(yàn)中我們將新西蘭大白兔制作脛骨慢性骨髓炎模型,然后對比蒸餾水煮沸、碘伏及敏感抗生素浸洗三種這不同處理方法后通過細(xì)菌學(xué)培養(yǎng)、組織學(xué)檢查等指標(biāo)的殺菌效果,本次試驗(yàn)的目的在于為今后臨床上在慢性骨髓炎殺菌方法的選擇上提供更優(yōu)異、有效、方便的方案,現(xiàn)將實(shí)驗(yàn)報(bào)告如下。方法:1兔脛骨慢性骨髓炎模型的制備。選取5月齡,體重1.5kg-2.5kg新西蘭大白兔50只,于清潔級環(huán)境中分籠術(shù)前適應(yīng)性飼養(yǎng),自由活動。3%戊巴比妥鈉按照1.0/kg的劑量注入耳緣靜脈進(jìn)行麻醉,麻醉滿意后,右脛骨內(nèi)側(cè)近端備皮,大小范圍約4cm,備皮完成后取右脛骨內(nèi)側(cè)縱切口,長約4cm,無菌手術(shù)刀切開皮膚和皮下組織,暴露右脛骨近端內(nèi)側(cè),鈍性剝離約2*2大小的骨膜,取用3mm鉆頭的電鉆,只穿透一側(cè)皮質(zhì),依此法在分離的骨膜處鉆9個等大骨洞,再用直徑1cm的磨鉆,制成一個約1*1cm的較為圓形的骨缺損即骨窗;然后用小刮匙分別沿骨窗遠(yuǎn)近端分別刮除部分松質(zhì)骨,完成后以0.9%生理鹽水5ml沖洗遠(yuǎn)近端髓腔。取用培養(yǎng)完畢后的金黃色葡萄球菌用接種環(huán)將菌體刮下,置于一次性無菌試管內(nèi),選取標(biāo)準(zhǔn)比濁管作為濃度標(biāo)準(zhǔn)參考物,參考6號標(biāo)準(zhǔn)管,用滴定管吸取0.9%生理鹽水予以滴定,制成濃度為1×106cfu/ml金黃色葡萄球菌液1ml,搖晃均一,用一次性滴定管吸取0.5ml的金黃色葡萄球菌液從骨窗內(nèi)向髓腔內(nèi)沿遠(yuǎn)近端注入,靜置5min,待菌液充分?jǐn)U散,再向骨窗內(nèi)沿遠(yuǎn)近端注射0.5ml的5%魚肝油酸鈉,用生理鹽水反復(fù)沖洗傷口后逐層縫合皮膚。術(shù)中死亡1只,術(shù)畢,待麻醉效果消失后回籠常規(guī)喂飼養(yǎng)4周。造模4周后分別從大體觀察、x線檢測及細(xì)菌培養(yǎng)三方面判斷50只兔子,是否形成骨髓炎。2確認(rèn)兔脛骨骨髓炎模型。試驗(yàn)后兔子均出現(xiàn)不同程度發(fā)熱,精神差,進(jìn)食少等表現(xiàn),均是骨髓炎后的臨床表現(xiàn),手術(shù)后4周內(nèi)因死亡2只未成功造模2只。術(shù)后4周,將存活的49只兔子,在攝x線片(42kv,4ma·s-1,距離1.0m)下攝片,發(fā)現(xiàn)47只兔右脛骨手術(shù)部位有慢性化膿性骨髓炎表現(xiàn),從肉眼觀察均呈現(xiàn)出局部軟組織腫脹,可伴有破潰,部分存在骨質(zhì)破壞,增生的骨痂,且有死骨形成;所有兔子取原手術(shù)切口4cm,取出小部分髓腔內(nèi)容物做病理學(xué)及細(xì)菌學(xué)檢測。綜合影像、病理及細(xì)菌學(xué)3方面做出判斷,其中細(xì)菌學(xué)檢測作為實(shí)驗(yàn)的金標(biāo)準(zhǔn)。檢測回報(bào)47只兔子均滿足要求,可作為兔脛骨骨髓炎模型做下一步實(shí)驗(yàn)。3比較三種不同殺菌方法的殺菌效果。取造模成功的兔子,將其隨機(jī)分成3組,分別截取病骨,然后編號,做不同處理。a組:截取病骨,用碘伏沖洗30分鐘;b組:蒸餾水煮沸30分鐘;c組:抗生素(對金黃色葡萄球菌敏感)清洗。三組均隨機(jī)選取處理過的骨病,置于標(biāo)準(zhǔn)培養(yǎng)基中培養(yǎng)24小時以做細(xì)菌學(xué)培養(yǎng)實(shí)驗(yàn)及組織學(xué)病理檢測。待結(jié)果有所回報(bào),匯總所有病例結(jié)果,以達(dá)到合理、可靠地比較三種方法殺菌效果的標(biāo)準(zhǔn)。結(jié)果:1大體觀察:三組處理方法處理后,均未出現(xiàn)骨質(zhì)破壞,均能保持較好的形態(tài),與原脛骨均有很好的形態(tài)相容性。2細(xì)菌學(xué)培養(yǎng):a組骨缺損周圍組織細(xì)菌培養(yǎng)結(jié)果顯示1例出現(xiàn)菌落,陽性率為7.00%,b組細(xì)菌培養(yǎng)結(jié)果提示4例陽性,陽性率為27.00%。c組細(xì)菌學(xué)培養(yǎng)提示7例陽性,陽性率為47.00%,兩兩比較差異均有顯著性。3病理學(xué)檢測:a處理組在he染色下結(jié)果顯示骨組織無水腫表現(xiàn),存在大片死骨,未見嗜酸性粒細(xì)胞等炎細(xì)胞存在,死骨呈片狀或條帶狀,且零星、無規(guī)律存在成骨母細(xì)胞;B處理組在HE染色下顯示為骨組織輕度水腫,無明顯死骨形成,可見少數(shù)嗜酸性粒細(xì)胞等炎細(xì)胞,且可見大量成骨細(xì)胞;C處理組在HE染色下顯示為骨及軟組織結(jié)構(gòu)較為混亂伴水腫,雖無明顯死骨形成,但可見嗜酸性粒細(xì)胞等炎細(xì)胞及成骨細(xì)胞。結(jié)論:蒸餾水煮沸、碘伏及敏感抗生素浸洗對兔脛骨慢性骨髓炎病骨均有一定的殺菌作用,且三種方法處理后依然能出色得保持原有的脛骨形態(tài)構(gòu)造,而蒸餾水煮沸不但能在短時間內(nèi)達(dá)到滅菌效果,而且仍有成骨細(xì)胞存在。
[Abstract]:Chronic osteomyelitis, chronic osteomyelitis, chronic osteomyelitis caused by high energy damage, such as trauma, traffic injury, especially open bone injury without thorough debridement or thorough debridement, is a difficult problem for Department of orthopedics physicians. The clinical treatment of chronic osteomyelitis is the main treatment of debridement. The sensitive antibiotics of 4-6 weeks of intramedullary cavity lavage and intravenous therapy can relieve some of the symptoms after the end of a course of treatment. It is often accompanied by symptoms such as low fever, dry throat, and general discomfort. Routine examination suggests that leukocytes are still at a high value and are clinically understandable as osteomyelitis. The recurrent attacks of osteomyelitis are accompanied by the serious drug resistance to antibiotics, resulting in the continuous escalation of antibiotics and the consequent continuous improvement of antibiotics. In this study, we made the New Zealand white rabbit model of the chronic osteomyelitis of tibia, and then compared the bactericidal effect of three different treatments, such as distilled water boiling, iodophor and sensitive antibiotic baptism, after three different treatments, such as bacteriological culture and histological examination. The purpose of the experiment is to provide a better, effective and convenient method for the clinical selection of chronic osteomyelitis in the future. Methods: 1 rabbit model of chronic osteomyelitis of tibia was prepared. 5 month old, 50 New Zealand white rabbits weight 1.5kg-2.5kg were selected for adaptive feeding in the clean grade environment. The free activity of.3% pentobarbital sodium was injected into the auricular vein at the dose of 1.0/kg. After the anesthesia was satisfied, the medial proximal end of the right tibia was about 4cm. The medial longitudinal incision of the right tibia was taken for a length of about 4cm. The skin and subcutaneous tissue were cut out by the aseptic scalpel, and the medial proximal end of the right tibia was exposed and the bone about 2*2 size of the bone was stripped blunt. The film, using a 3mm drill, penetrates only one side of the cortex, and then drills 9 large bone holes at the separated periosteum, and then the diameter 1cm's drill is used to make a more circular bone defect, the bone window of about 1*1cm, and then scrape off some of the cancellous bone with a small curet along the far end of the bone window and then rinse the far and near end with 0.9% physiological saline 5ml. The Staphylococcus aureus was scraped by inoculation ring and placed in a single sterile test tube. The standard turbid tube was selected as the reference material of the concentration standard, and 0.9% normal saline was titrated with the burette to be titrated with the burette 6. The concentration of Staphylococcus aureus was 1 * 106cfu/ml Staphylococcus liquid 1ml. 0.5ml Staphylococcus aureus liquid was injected into the intramedullary cavity from the bone window into the proximal and proximal ends of the Staphylococcus aureus from the bone window, then 5min was inserted, the bacterial liquid was diffused fully, then the 5% sodium cod liver oil was injected along the far and near end of the bone window, and the wound was sutured after the wound was repeatedly washed with saline. During the operation, 1 were dead, after the operation, the anesthesia effect disappeared after the effect disappeared. The cage was fed for 4 weeks. After 4 weeks, 50 rabbits were judged from three aspects of gross observation, X-ray examination and bacterial culture. Whether osteomyelitis.2 was formed to confirm the rabbit tibial osteomyelitis model. After the experiment, the rabbits showed different degrees of fever, poor spirit, and less eating, all were the clinical manifestations of osteomyelitis, and 2 died in 4 weeks after the operation. 4 weeks after operation, 49 rabbits survived in 49 rabbits were photographed on X-ray films (42KV, 4mA s-1, and distance 1.0m). The results showed that there were chronic suppurative osteomyelitis in the right tibia of 47 rabbits. All rabbits were taken the original surgical incision 4cm, take out a small part of the marrow cavity content to do pathological and bacteriological examination. Comprehensive imaging, pathology and bacteriology 3 aspects to make a judgement, bacteriological detection as the gold standard of the experiment. 47 rabbits were satisfied with the detection return, can be used as the rabbit tibial osteomyelitis model for the next experiment.3 comparison three kinds of no The bactericidal efficacy of bactericidal methods. The rabbits were randomly divided into 3 groups, which were randomly divided into 3 groups, and then the diseased bone was intercepted and then numbered, and the different treatment groups were treated for 30 minutes by intercepting the diseased bone and rinsed with Iodophor; group B: distilled water boiling for 30 minutes; group C: antibiotics (sensitive to Staphylococcus aureus). The three groups were randomly selected to select treated bone disease randomly and placed at random, placed at random The standard culture medium was cultivated for 24 hours to do bacteriological culture experiment and histopathological examination. The results were rewarded and all the results were collected in order to achieve a reasonable and reliable comparison of the standard of three methods of sterilization effect. Results: 1 gross observation: after treatment of three groups, no bone destruction was found, and good form could be maintained. There were good morphological compatibility.2 bacteriological culture with the original tibia: the results of bacterial culture in the tissue surrounding the bone defect in group A showed that 1 cases were colonies, the positive rate was 7%, and 4 cases were positive in group B, and the positive rate was 7 positive in group 27.00%.c bacteriology, the positive rate was 47%, and the difference of the 22 difference had significant.3 pathology. Study test: a treatment group showed no edema in bone tissue in HE staining, there were large dead bone, no eosinophil and other inflammatory cells existed, dead bone was flake or strip, and sporadic and irregular in osteoblast; B treatment group showed mild edema in bone group, no obvious bone formation, and few eosinophils in B treatment group. A large number of osteoblasts were found in the granulocyte, and a large number of osteoblasts were seen. The C treatment group showed that the bone and soft tissue were more chaotic and oedema with HE staining, although there was no obvious dead bone formation, but eosinophils and osteoblasts were visible. Conclusion: boiling water, iodophor and sensitive antibiotic Baptist in rabbit bone of chronic osteomyelitis of tibia It has a certain bactericidal effect, and the three methods can still be treated well to maintain the original shape and structure of the tibia, and the boiling water can not only achieve the sterilization effect in a short time, but still there are osteoblasts.
【學(xué)位授予單位】：承德醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R681.2

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