探讨蒸馏水煮沸、碘伏及敏感抗生素浸洗对兔胫骨慢性骨髓炎病骨的杀菌效果比较

发布时间：2018-05-28 21:49

本文选题：慢性骨髓炎 + 兔　；参考：《承德医学院》2017年硕士论文

【摘要】：目的:慢性骨髓炎指骨组织的慢性感染,由外伤、交通伤等高能量损伤特别是开放性骨损伤未经彻底清创或虽经彻底清创但创伤及污染较重或由手术时无菌操作不严格所引起,是困扰骨科医师的难题之一。慢性骨髓炎因其死骨形成、窦道、死腔及病变肉芽组织的长期存在,治疗较困难。目前对慢性骨髓炎的临床治疗措施是清创术的主要治疗方法,髓腔灌洗和静脉治疗4-6周的敏感抗生素,在一个疗程结束后通常能缓解部分症状,红肿热痛等炎症表现可有明显减轻,但患者常常伴有低烧,咽喉干燥,全身不适等症状,常规检查提示白细胞仍处在较高值,在临床上可理解成骨髓炎复发。骨髓炎反复发作的同时也伴随着人体对抗生素严重耐药的现象,造成的结果为必然抗生素的不断升级,并由此带来的不断升级抗生素[4]以及再次升级再次耐药的恶性循环。该研究实验中我们将新西兰大白兔制作胫骨慢性骨髓炎模型,然后对比蒸馏水煮沸、碘伏及敏感抗生素浸洗三种这不同处理方法后通过细菌学培养、组织学检查等指标的杀菌效果,本次试验的目的在于为今后临床上在慢性骨髓炎杀菌方法的选择上提供更优异、有效、方便的方案,现将实验报告如下。方法:1兔胫骨慢性骨髓炎模型的制备。选取5月龄,体重1.5kg-2.5kg新西兰大白兔50只,于清洁级环境中分笼术前适应性饲养,自由活动。3%戊巴比妥钠按照1.0/kg的剂量注入耳缘静脉进行麻醉,麻醉满意后,右胫骨内侧近端备皮,大小范围约4cm,备皮完成后取右胫骨内侧纵切口,长约4cm,无菌手术刀切开皮肤和皮下组织,暴露右胫骨近端内侧,钝性剥离约2*2大小的骨膜,取用3mm钻头的电钻,只穿透一侧皮质,依此法在分离的骨膜处钻9个等大骨洞,再用直径1cm的磨钻,制成一个约1*1cm的较为圆形的骨缺损即骨窗;然后用小刮匙分别沿骨窗远近端分别刮除部分松质骨,完成后以0.9%生理盐水5ml冲洗远近端髓腔。取用培养完毕后的金黄色葡萄球菌用接种环将菌体刮下,置于一次性无菌试管内,选取标准比浊管作为浓度标准参考物,参考6号标准管,用滴定管吸取0.9%生理盐水予以滴定,制成浓度为1×106cfu/ml金黄色葡萄球菌液1ml,摇晃均一,用一次性滴定管吸取0.5ml的金黄色葡萄球菌液从骨窗内向髓腔内沿远近端注入,静置5min,待菌液充分扩散,再向骨窗内沿远近端注射0.5ml的5%鱼肝油酸钠,用生理盐水反复冲洗伤口后逐层缝合皮肤。术中死亡1只,术毕,待麻醉效果消失后回笼常规喂饲养4周。造模4周后分别从大体观察、x线检测及细菌培养三方面判断50只兔子,是否形成骨髓炎。2确认兔胫骨骨髓炎模型。试验后兔子均出现不同程度发热,精神差,进食少等表现,均是骨髓炎后的临床表现,手术后4周内因死亡2只未成功造模2只。术后4周,将存活的49只兔子,在摄x线片(42kv,4ma·s-1,距离1.0m)下摄片,发现47只兔右胫骨手术部位有慢性化脓性骨髓炎表现,从肉眼观察均呈现出局部软组织肿胀,可伴有破溃,部分存在骨质破坏,增生的骨痂,且有死骨形成;所有兔子取原手术切口4cm,取出小部分髓腔内容物做病理学及细菌学检测。综合影像、病理及细菌学3方面做出判断,其中细菌学检测作为实验的金标准。检测回报47只兔子均满足要求,可作为兔胫骨骨髓炎模型做下一步实验。3比较三种不同杀菌方法的杀菌效果。取造模成功的兔子,将其随机分成3组,分别截取病骨,然后编号,做不同处理。a组:截取病骨,用碘伏冲洗30分钟;b组:蒸馏水煮沸30分钟;c组:抗生素(对金黄色葡萄球菌敏感)清洗。三组均随机选取处理过的骨病,置于标准培养基中培养24小时以做细菌学培养实验及组织学病理检测。待结果有所回报,汇总所有病例结果,以达到合理、可靠地比较三种方法杀菌效果的标准。结果:1大体观察:三组处理方法处理后,均未出现骨质破坏,均能保持较好的形态,与原胫骨均有很好的形态相容性。2细菌学培养:a组骨缺损周围组织细菌培养结果显示1例出现菌落,阳性率为7.00%,b组细菌培养结果提示4例阳性,阳性率为27.00%。c组细菌学培养提示7例阳性,阳性率为47.00%,两两比较差异均有显著性。3病理学检测:a处理组在he染色下结果显示骨组织无水肿表现,存在大片死骨,未见嗜酸性粒细胞等炎细胞存在,死骨呈片状或条带状,且零星、无规律存在成骨母细胞;B处理组在HE染色下显示为骨组织轻度水肿,无明显死骨形成,可见少数嗜酸性粒细胞等炎细胞,且可见大量成骨细胞;C处理组在HE染色下显示为骨及软组织结构较为混乱伴水肿,虽无明显死骨形成,但可见嗜酸性粒细胞等炎细胞及成骨细胞。结论:蒸馏水煮沸、碘伏及敏感抗生素浸洗对兔胫骨慢性骨髓炎病骨均有一定的杀菌作用,且三种方法处理后依然能出色得保持原有的胫骨形态构造,而蒸馏水煮沸不但能在短时间内达到灭菌效果,而且仍有成骨细胞存在。
[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.
【学位授予单位】：承德医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R681.2

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