【作者】 冯文东；

【导师】 倪才方；

【作者基本信息】 苏州大学 ， 影像医学与核医学， 2009， 硕士

【摘要】 目的探讨磷酸钙骨水泥(CPC)和聚甲基丙烯酸甲酯骨水泥(PMMA)对实验兔椎旁转移瘤的细胞毒作用,研究动物椎旁转移瘤模型代替脊柱肿瘤模型进行实验研究的可行性。材料及方法1. CT定位下经皮穿刺瘤块推送法建立实验兔VX2椎旁转移瘤模型。15天后MRI检查模型建立情况。2.模型兔随机分为3组:①对照组:模型兔无任何处理。②PMMA组:模型兔肿瘤内注入PMMA骨水泥。③CPC组:模型兔肿瘤内注入CPC骨水泥。3.对照组不做处理,CPC组及PMMA组在CT定位下向椎旁转移瘤中心分别注入CPC或PMMA骨水泥1.0ml。骨水泥注入后每组分别于1天、7天、14天后处死5只荷瘤兔,取出肿瘤组织,10%甲醛固定。标本切取两部分:骨水泥表面肿瘤组织、骨水泥远端肿瘤组织(对照组取肿瘤中心和边缘),经冲洗、脱水、透明、包埋、制成蜡块后切片。4.①骨水泥注入后每天观察动物生理状态;②HE染色法观察骨水泥表面及边缘肿瘤细胞形态变化及坏死情况;③TUNEL细胞原位凋亡检测,观察肿瘤细胞凋亡并计算凋亡指数,所得数据采用SPSS13.0统计软件包行统计学分析。结果①模型动物均能很好的耐受手术,骨水泥注入后成团状,无明显的渗漏。术后实验动物无明显的生理不适,无精神萎靡、嗜睡、拒食等现象;②HE染色结果显示PMMA组术后骨水泥表面肿瘤细胞发生不同程度的坏死,细胞出现典型的核浓缩、核分裂、核溶解、胞浆溶解红染,在坏死的肿瘤组织中可见少许残存的肿瘤细胞,尤以骨水泥远端较多见,7天及14天后骨水泥表面肿瘤细胞较第1天有明显增生,边缘及周边肿瘤细胞增生活跃;对照组和CPC组各时间段均显示骨水泥表面和边缘肿瘤组织增生活跃;③TUNEL细胞原位凋亡检测结果显示:各时间点内PMMA组骨水泥表面肿瘤细胞凋亡较周边多,治疗后第1天差异较为明显,以后逐渐减少。对照组和CPC组骨水泥表面及周边细胞凋亡不明显。PMMA组各时间点骨水泥表面及周边肿瘤细胞凋亡较对照组和CPC组明显增加,两者比较有统计学意义(P<0.05)。CPC组和对照组则无明显差异,两者比较无统计学意义(P>0.05)。结论PMMA具细胞毒性,对活体肿瘤细胞具有一定的杀伤作用,CPC则没有明显的肿瘤杀伤作用。实验兔椎旁转移瘤模型建立比较容易,可能作为动物脊柱肿瘤模型进行有关实验研究。更多还原

【Abstract】 ObjectiveTo investigate the anti-tumor effect of two kinds of bone cements(CPC and PMMA) in rabbit paravertebral metastatic tumors. To research the feasibility of the paravertebral me-tastatic tumor model to replace the spine tumor model.Methods and methods1. Transplantable tumor model were constructed by paracentesis under the CT guiding. The rabbits were checked after 15 days by MRI. 2. The rabbit models were randomly divided into 3 groups: Control Group-nothing was injected with the rabbits, PMMA Group-PMMA 1 ml was injected to the center of the tumor, CPC Group-CPC 1 ml was injected to the center of the tumor. 3. The rabbits were executed at 1d、7d and 14d after bone cement were injected. The tissues of tumor were washed、dehydrated、permeabilized、embeded and cut into section after tumor tissue dissection. 4.①The model rabbits were observed every day after operation;②The morphological change and necrosis of tumor cells near and away from the bone cements were observed by HE stain③Apoptotic feature and apoptotic rate were analyzed by TUNEL apoptosis measurement. The data were statistically analyzed by SPSS13.0 statistical bag.Results①All model animals can tolerate the operation well. Bone cement was agglomerate in tumor, and was not found cement leakage. Physiological discomfort, psychical dispirited, drowsiness and fastidium were not observed after operation on rabbits;②HE stain results suggested that there was typical nuclei concentration、nuclei cleavage、nuclei dissolution and necrosis of tumor cells near the PMMA bone cements. Few of tumor cells could found in necrosis tissues. Until at 7 days and 14 days after operation, the tumor cells near bone cement were significantly proliferated. But tumor cells significantly proliferated were found on control group and CPC group at all time.③TUNEL apoptosis examination showed that PMMA could induce apoptosis on tumor cells near bone cements,and the PMMA’s apoptotic effect was reduced gradually. At the different time after treatment, the apoptosis index (AI, %) of tumor tissue in the PMMA group was higher than that of in control group and CPC group. The difference of apoptosis index PMMA group compared with other two group has statistical significance (p<0.001). There is no statistical signifi-cance of CPC group and control group (p>0.001).Conclusion:The results suggest that PMMA has a significantly cytotoxic and anti-tumor effect, and CPC haven’t the effect. The method establishing rabbit paravertebral metastatic tumor model is easy, which can replace the spine tumor model partially.更多还原