自制改良穿刺装置的动物模型建立及免疫组化研究

发布时间：2018-03-26 06:51

本文选题：VX2肿瘤细胞　切入点：动物模型　出处：《承德医学院》2017年硕士论文

【摘要】：目的:通过实验设立兔VX2骨转移瘤动物模型,观察确定适合穿刺装置穿刺活检的时间,明确适宜的穿刺时间后,通过PCNA免疫组化技术,验证自制改良穿刺装置的效果。方法:实验所应用的动物为人工养殖的家兔,第一部分所用家兔数量为25只,第二部分35只,实验所用的各种设备来自于承德医学院附属医院实验室,所用的肿瘤细胞以及试剂采购于试剂公司。1、第一部分为制造一种适合本实验的模型,观察肿瘤模型随着时间的推移而进展的各期变化,大体明确肿瘤生长需要的时间;采购VX2肿瘤细胞,植入家兔后侧肢体的胫骨近端,通过将肿瘤细胞悬液置入家兔胫骨骨孔内的方法建立模型;随机抽取5只为对照组,其余20只以双侧后肢为实验肢体,左侧肢体采用肿瘤组织悬液植入法建立兔VX2骨转移瘤模型(左侧组),右侧肢体植入生理盐水(右侧组),分别于种植后每周通过影像学及病理学检查观察肿瘤生长情况,进而明确VX2肿瘤进展各期大体时间。2、第二部分为验证自制骨肿瘤穿刺装置效果的实验,在已成熟的制造家兔VX2胫骨骨肿瘤模型以及明确髓腔内瘤体成型但尚未突破骨皮质这一阶段的时间基础上,通过PCNA免疫组化技术来验证该装置的效果;选购35只实验兔,按上述方法制造家兔VX2胫骨骨肿瘤模型,双侧后肢均植入肿瘤组织,随机选出5只,作为对照组,其余30只分为左侧组及右侧组,左侧组应用自制改良穿刺针于皮质骨上打孔后封堵,右侧组应用传统的穿刺针进行打孔(此时穿刺针于皮质上打孔的位置应与造模时植入肿瘤组织的孔相对,避免造模时形成的孔道影响实验效果),两组穿刺术完成后仔细缝合,同样层数无菌敷料包扎(3层),术后于实验后次日观察两组实验动物手术部位渗出情况,进而明确两组实验动物术后手术部位出血情况有无差异;将两组实验动物于穿刺术完成后第10天处死,获取穿刺针遗留孔周围的软组织,通过PCNA免疫组化技术查看穿刺针遗留的孔周围有无肿瘤细胞扩散,对比两种装置的效果。结果:1、第一部分制造成功家兔VX2胫骨骨肿瘤模型,通过观察影像学及病理学检查,确定植入肿瘤组织后第2周末,即第14天可见髓腔内成型瘤体,并且未侵犯皮质,左侧组植入肿瘤细胞后1~2周肿瘤组织于髓内生长,第3周开始肿瘤组织髓内生长并侵犯部分骨皮质,第4周末肿瘤组织突破骨皮质侵犯周围软组织。2、穿刺术实施后的次日见30只实验动物中,左侧组手术部位敷料可见血性渗出者1只,余下者敷料均干燥无渗出;右侧组手术部位有25只可见大量血性渗出,2只少量血性渗出,3只敷料干燥无渗出;第10天处死所有实验动物,获取穿刺针所形成的孔道周围软组织行PCNA免疫组化技术处理,左侧组即应用自制改良穿刺针组穿刺孔道周围软组织无肿瘤细胞侵及,而应用传统穿刺针组穿刺孔周围可见肿瘤细胞侵及。结论:成功地建立了一种兔VX2骨肿瘤模型,此模型可很好的应用于骨肿瘤方面各类研究的动物实验中,利用自制的穿刺装置实施穿刺术,降低了穿刺术实施后各项并发症的发生率,证明自制改良穿刺装置可以有效地应用于肿瘤组织的穿刺活检,是一种新型的有效的装置和技术。这个装置的特点为组合式的各部器械,通过穿刺针穿过骨皮质后,利用装置外侧的套管来阻断骨皮质与骨髓内的肿瘤细胞,同时使髓腔与外界形成通道,在我们获取需要的足够组织之后,通过套管将装置的关键物即封闭栓置入穿刺孔内,进而避免了骨髓腔内的肿瘤细胞通过穿刺孔扩散,并且此方法创伤较传统穿刺针无增大,但对于穿刺后各项并发症的预防却有了显著地提高。该装置可在病理检查未明确之前,尽量减少肿瘤组织的扩散,同时可减少穿刺部位的出血等并发症发生,降低血肿形成的几率,对于临床工作有很大的意义。
[Abstract]:Objective: to set up animal model of rabbit VX2 bone metastases, observe the biopsy puncture device to determine the appropriate time, clear suitable time of puncture, by PCNA immunohistochemical technique, test the effect of modified puncture device. Methods: the application of the animal for artificial breeding of rabbits, the first part of the number of rabbits for the 25, second part 35, all kinds of equipment used in the experiment from the laboratory of Affiliated Hospital of Chengde Medical College, the tumor cells and reagent to the reagent company.1, the first part is to make a suitable experimental model, to observe the tumor model with the passage of time and the progress of each stage of change, generally clear tumor growth needs the time of purchase; VX2 tumor cells implanted in the posterior limb of the proximal tibia, the tumor cell suspension liquid into rabbit tibia bone hole is built with the model; The machine selected 5 rats as the control group, the remaining 20 only to the hind limbs of experimental limb, left limb tumor metastasis by established rabbit model of VX2 bone suspension implantation (left group), the right limb was filled with saline (right), respectively, after planting in a week by imaging and pathological examination to observe the growth situation the tumor, and then clear the approximate time of tumor progression of VX2.2, the second part is to verify the self-made bone tumor puncture device experiment in time based manufacturing VX2 has mature rabbit tibial bone tumor model and clear intramedullary tumor forming but not breaking the cortical bone in this stage, by PCNA immunohistochemical technique to verify the efficacy of the device; choose 35 rabbits, rabbit VX2 bone tumor model made by the above method, the hind limbs were implanted into the tumor tissues, 5 rats were randomly chosen as the control group, the remaining 30 were divided into the left The left and right group group group using self-made puncture needle punched in cortical bone after the closure, the needle right by applying the traditional drilling (the position of the puncture needle from the cortical perforating and when modeling implanted tumor tissue relative to avoid the formation of holes, when modeling the effects of pore effect), puncture in two groups after careful suture, the same number of sterile dressing (3), after the operation on the day after the experimental observation of the two groups of experimental animal surgical site exudation, and then clear the surgery site bleeding in two groups of experimental animal postoperative difference; two groups of experimental animal to puncture tenth days after death, get the needle from the soft tissue around the hole, by PCNA immunohistochemical techniques around the needle hole is left to see without the spread of tumor cells, comparing the two kinds of device. Results: 1, the first part of the manufacture of successful VX2 rabbit tibia Bone tumor model by observing the imaging and pathological examination, determine the tumor tissue second weeks after implantation, fourteenth days intramedullary tumor forming visible, and not involving the cortex, the left group at 1~2 weeks after implantation of tumor cells in tumor tissue growth in intramedullary tumor tissue third weeks intramedullary bone growth and invasion at the end of the fourth breakthrough cortex, tumor invasion of cortical bone soft tissue around.2, puncture time is after the implementation of the 30 experimental animal, dressing group left surgical site visible blood exudation in 1 cases, the rest of them were dressing dry without exudation; right surgical site 25 showed a large amount of blood exudation, 2 is only a small amount of blood 3 dry dressing exudation exudation; tenth days all the experimental animal, get the puncture needle formed around the channels of soft tissue by PCNA immunohistochemical technique, the left group application of modified puncture needle puncture group No way of soft tissue around the tumor cells invasion, and the application of traditional needle puncture holes around the tumor cells invasion. Conclusion: a rabbit VX2 bone tumor model was successfully established, and this model can be well applied in animal experiment of bone tumor research, the implementation of self puncture puncture device system and reduce the complications of puncture after the implementation of the incidence of biopsy proved that modified puncture device can be effectively applied to the tumor tissue, is a new and effective device and technology. The utility model is a combination of the device through the puncture needle through the cortical bone, bone block bone marrow tumor cells in the cortex and the outer sleeve device, the medullary cavity and the outside channel formed after we get enough tissue to, through the casing will be key device that closed bolt placement wear Puncture, and avoid the bone marrow tumor cells through the puncture hole diffusion, and this method is better than the traditional wound puncture needle does not increase, but for the prevention of complication has been significantly improved. The device can be before pathological examination is not clear, try to reduce the spread of tumor tissue, and can reduce hemorrhage the occurrence of complications such as puncture, reduce the risk of hematoma formation, has a great significance for clinical work.

【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】：R-332;R738

【参考文献】