假膜成纤维细胞诱导裸鼠骨溶解动物模型的实验研究

发布时间：2018-04-28 17:23

  本文选题：成纤维细胞 + 人工关节置换　； 参考：《上海交通大学》2007年硕士论文

【摘要】： 【目的】研究应用假膜成纤维细胞构建人工关节周围骨溶解裸鼠体内模型的可行性。 【方法】本实验于2006年5月至2007年1月于上海市第六人民医院中心实验室和动物实验室完成,取裸鼠(SCID小鼠)15只,随机分为三组,每组5只;分别为假膜成纤维细胞组(组I),关节囊成纤维细胞组(组II),对照组(组III),假膜成纤维细胞组从人工关节无菌性松动翻修术中取得的假膜组织,分离出成纤维细胞,经过3～4次传代后,应用骨水泥刺激成纤维细胞后植入裸鼠的颅盖骨上;关节囊成纤维细胞组从初次人工关节置换术中取出部分关节囊组织,分离出成纤维细胞经过与组I同样的处理后植入裸鼠的颅盖骨上,对照组为假手术组;两周后处死裸鼠,取出颅盖骨脱钙处理后切片,应用影像软件分析裸小鼠颅中缝的骨溶解情况,定量分析骨溶解的面积以判断骨吸收的程度。 【结果】假膜组织和关节囊组织中分离出的细胞经过鉴定表达成纤维细胞相关抗原(脯氨酸-4-羟化酶),经RT-PCR检测发现假膜成纤维细胞高表达RANKL与OPG的mRNA,假膜组织的成纤维细胞植入裸鼠颅盖骨后可以从H-E染色的切片中观察到裸鼠颅盖骨骨吸收,经过IPP软件分析测量后假膜成纤维细胞组平均骨吸收面积为0.23±0.030 mm2(P0.05),明显高于关节囊成纤维细胞组(组Ⅱ)(0.083±0.013mm2)和对照组(组Ⅲ)(0.078±0.009 mm2)。 【结论】从假膜中分离成纤维细胞可操作性强,假膜成纤维细胞植入裸鼠后可以成功诱导骨溶解形成,可重复性强,可以作为研究人工关节周围骨溶解的动物模型。
[Abstract]:[objective] to study the feasibility of pseudomembrane fibroblast to construct osteolysis model of artificial joint in nude mice. [methods] from May 2006 to January 2007, the experiment was carried out in the Central Laboratory and Animal Laboratory of Shanghai sixth people's Hospital. Fifteen SCID mice were randomly divided into three groups with 5 mice in each group. The pseudomembrane fibroblasts group (group I), the joint capsule fibroblast group (group II), the control group (group II), and the pseudomembranous fibroblast group (group II) were isolated from the pseudomembranous tissue obtained from the artificial joint aseptic loosening revision. After 34 passages, bone cement was used to stimulate fibroblasts and implanted into the skull of nude mice. After the same treatment as group I, the fibroblasts were implanted into the skull of nude mice, the control group was sham-operated group, the nude mice were killed two weeks later, the skull cap was decalcified and then sliced. The osteolysis of cranial suture in nude mice was analyzed by image software, and the area of osteolysis was quantitatively analyzed to judge the degree of bone resorption. [results] the cells isolated from pseudomembranous tissue and articular capsule were identified to express fibroblast-associated antigen (proline -4-hydroxylase). By RT-PCR detection, it was found that pseudofibroblasts expressed RANKL and OPG mRNAs and pseudomembrane tissues. After implantation of fibroblasts into the skull of nude mice, bone resorption of skull cap of nude mice could be observed from H-E stained sections. The average bone resorption area of the pseudomembranous fibroblasts group was 0.23 卤0.030 mm ~ (-2) (P 0.05), which was significantly higher than that of the articular capsule fibroblast group (group 鈪，

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