假膜成纤维细胞诱导裸鼠骨溶解动物模型的实验研究-外科学专业论文.docxVIP

假膜成纤维细胞诱导裸鼠骨溶解模型的实验研究 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。 【关键词】成纤维细胞，人工关节置换，骨溶解，动物模型 ARTHROPLASTY MEMBRANE-DERIVED FIBROBLASTS INDUCE OSTEOLYSIS IN A SEVERE COMBINED IMMUNODEFICIENT MICE MODEL ABSTRACT Aim:To develop a SCID(severe combined immunodeficient mice) model of osteolysis induced by Arthroplasty membrane-derived fibroblasts. Methods:the experiment was carried out in the Laboratory Animal Center of shanghai Sixth People’s Hospital from match 2006 to april 2007. Fifteen SCID mice were select and randomly divided into 3 groups:Fibroblasts were isolated from periprosthetic tissue during the revision and articular capsule during the primary THA.Then the fibroblasts were implanted onto the calvarial of the SCID mice,which was divided into three groups: group I:Arthroplasty membrane-derived fibroblasts group,group II :capsule derived fibroblasts group and group III:control group. groupⅠ:The fibroblasts were received from periprosthetic tissue ，after passaging the cells 3-4 times ,the fibroblasts were implanted onto the calvarial of the SICD mice,group Ⅱ:the fibroblasts were received from articular capsule ,group Ⅲ underwent sham surgery only.After two weeks, calvaria were processed for histomorphometry and the bone resorption area was calculated automatically by the software. Result: These ce