兔同种异体骨异位再血管化的实验研究
发布时间：2018-03-19 17:08
本文选题：同种异体骨 切入点：大段骨缺损 出处：《新乡医学院》2015年硕士论文 论文类型：学位论文
【摘要】：1 背景在机械、工业、交通日渐发达的今天,各种高能量损伤越来越多,程度越来越复杂。小腿开放性粉碎骨折发生率最高,术后骨感染率较高,且易转化为慢性骨感梨[1]。彻底去除感染骨段及其周围病灶是控制感染的重要手段,但可能造成骨缺损甚至大段骨缺损,且控制局部创面感染往往还需经过漫长的治疗过程。创伤后感染性大段骨缺损是骨科难题,同种异体骨用于感染性大段骨缺损的修复尚未见报道,能否将其用于感染性大段骨缺损的治疗?我们设想将同种异体骨段置于体内血供丰富的部位,使其完成再血管化,待创面感染彻底控制后,再将其带血管蒂的血管化同种异体骨段移植修复骨缺损。2 目的以中国白兔为实验对象,将制备好的同种异体骨段置于兔血运丰富的隐动脉处,探索其完成再血管化的时间。3 方法3.1同种异体骨的制备选取10只中国白兔,空气栓塞耳缘静脉杀死白兔,按照无菌手术的方法取出兔胫骨,把周围的肌肉及骨膜剔除,截成1.5cm长度的骨段,把骨髓组织用小刮匙彻底刮除,用无菌生理盐水清洗干净,浸泡于10%的庆大霉素中30分钟,用灭菌好的双层血浆袋密封包装,经过辐照灭菌后,放入冰箱4。冷藏12h,-30°冷冻12h,在-80。的深低温冰箱3周后使用,可以去除更多的抗原。3.2同种异体骨再血管化以健康成年中国白兔为研究对象。把60只白兔按随机数字表法随机分为两组,其中实验组和对照组各30只。实验组把已经准备好的长1.5cm的异体骨段放置在兔大腿股直肌和股内侧间隙隐动脉处,用1.Omm.克氏针把骨段固定在股骨内侧,对照组为自体的胫骨段,长度和实验方法与实验组相同,术后4周,8周,12周分别从肉眼大体标本观察、免疫组化切片检测血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)和血管内皮细胞中CD34蛋白表达,行统计学处理。4结果实验组和对照组部分兔手术部位感染,不包括在统计标准内,补充空缺。VEGF以新生的、未成熟的血管周围,软骨细胞、成骨细胞以及未分化的间叶细胞周围最多,术后4、12周,VEGF的表达量实验组和对照组比较无差异,8周,与对照组相比,实验组VEGF的表达有显著统计学差异,两组纵向比较,8周多于4、12周。CD34蛋白以哈弗管周围的血管内皮细胞中最明显。术后4周,CD34阳性血管两组比较有显著差异,8、12周,实验组与对照组比较,无显著差异。实验组和对照组CD34阳性血管8周多于4、12周。术后4周,实验组和对照组都有较少的纤维组织膜包裹骨段、填充髓腔,与骨段结合松散,容易剥离,骨表面无明显吸收陷窝,出现光泽。术后8周,两组骨段上的纤维组织膜变厚,与骨段结合紧密,不易分开,骨段表面不平整,有吸收陷窝,出现光泽。术后12周两组骨段上包裹的纤维组织膜更厚,与骨段结合更紧密,骨表面的吸收陷窝更多,光泽更明显。5结论兔大段同种异体胫骨段在血运丰富的隐动脉处8周能够实现再血管化,时间与自体骨段无明显差异。
[Abstract]:1 background in mechanical, industrial, traffic increasingly developed today, a variety of high energy injury more and more, more and more complex. The highest incidence of open comminuted fracture, bone infection rate after operation, and is easily transformed into chronic skinny pear [1]. to remove the infection of bone and peripheral lesions is an important means to control infection may, but bone defect even large bone defect caused by wound infection, and control of local needs through the long course of treatment. After trauma infection of large bone defects is a department of orthopedics problem, allograft bone for repair of infected large bone defect has not been reported, whether it can be used for the treatment of infected bone defect? We assume the allogeneic bone in vivo and abundant blood supply parts to complete revascularization, to completely control the wound infection, then the vascular pedicled vascularized allograft bone transplantation Objective to repair bone defect.2 China rabbits, saphenous artery will be for allogeneic bone in rabbit blood prepared supplyrich, explore its completion time.3 method of revascularization of 3.1 allograft preparation select 10 China rabbit ear vein air embolism rabbits were killed, according to the method aseptic operation out of rabbit tibia, the surrounding muscle and periosteum removed, cut into bone segment length 1.5cm, the bone marrow tissue with small curette curettage, with sterile saline washed, soaked in 10% gentamicin in 30 minutes, sealed with double plasma sterilization bag, after irradiation after sterilization the refrigerator 4. refrigerated 12h, -30 ~ 12h in deep frozen, low temperature refrigerator -80. after 3 weeks of use, can remove the.3.2 antigen of allogeneic bone more revascularization in healthy adult rabbits were China as the research object. The 60 rabbits at random Randomly divided into two groups, the experimental group and the control group with 30 rats in each experimental group. The allograft bone is ready for the long 1.5cm placed in the rabbit thigh of rectus femoris and vastus medial saphenous artery, 1.Omm. Kirschner wire to bone fixation in the medial femoral, tibial autogenous control group section, length and experimental methods and the same as the experimental group, 4 weeks, 8 weeks after surgery, 12 weeks respectively from specimen observation, immunohistochemistry detection of vascular endothelial growth factor (vascular endothelial, growth factor, VEGF) expression of CD34 protein and vascular endothelial cells, statistical treatment results of.4 experimental group and control group rabbit surgical site infection, not included in the statistical standard, add.VEGF to new vacancies, around the immature vascular into cartilage cells, bone cells and the most undifferentiated mesenchymal cells around 4,12 weeks after operation, the expression of VEGF in the experimental group 鍜屽鐓х粍姣旇緝鏃犲樊寮，
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