腹腔内置疝修补补片动物实验的病理评价

目的以满足《腹腔内置疝修补补片动物实验技术审查指导原则》要求为基础,探讨客观、准确的病理检测指标和评价方法。 方法选取两种复合疝修补补片,分别植入犬腹腔内左右两侧腹壁。12周后将补片植入区域连同周围正常组织取材,采用免疫组织化学及免疫荧光染色方法评价新生腹膜比例、炎性反应;采用Masson染色方法检测纤维化程度;采用HE染色方法检测新生血管和脂肪浸润情况。 结果（1）Cytokeratin和Vimentin两种抗体均可以对间皮细胞进行识别,有助于对新生腹膜层长度的测量,Cytokeratin特异性更高;（2）应用抗体CD20、CD3和CD68免疫组化染色可以高效地检测出组织中B淋巴细胞、T淋巴细胞和巨噬细胞,辅助细胞计数和炎性反应评分;（3）CD34免疫组化染色观察新生血管情况并计算新生血管密度;（4）HE染色可以直接观察脂肪细胞浸润情况并进行半定量评分;（5）Masson染色可以清晰直观地反应胶原纤维沉积情况,对纤维沉积区域厚度进行测量、比较有辅助作用;（6）2组补片的所有观察指标通过定量和半定量方法进行描述,随后进行统计分析,结果显示所有指标间均无统计学差异（P>0.05）。 结论本研究采用的检测方法能对补片植入后的局部反应相关指标进行直接、准确的定位和量化,基本满足《腹腔内置疝修补补片动物实验技术审查指导原则》的要求,对今后开展相关实验具有一定的参考价值。

Pathological evaluation of animal experiments with intraperitoneal abdominal hernia repair

ObjectiveTo explore objective and accurate pathological detection indexes and evaluation methods based on the requirements of "Guidelines for technical review of intraperitoneal hernia repair mesh in animal experiments" . MethodsTwo kinds of composite hernia patches were implanted in the abdominal wall of dogs. After 12 weeks, the patch area and surrounding normal tissues were sampled, and the proportion of peritoneal mesothelial cells, the inflammatory response, and neovascularization were evaluated by immunohistochemistry and immunofluorescence staining. The degree of fibrosis was detected by Masson staining. Fat infiltration was detected by HE staining. Results(1) Both Cytokeratin and Vimentin could accurately identify mesothelial cells, which is helpful to measure the length of mesothelial cell layer; (2) Using antibodies CD20, CD3 and CD68 could effectively detect B lymphocytes, T lymphocytes and macrophages in tissues; (3) Using CD34 antibody could be used to observe the situation and calculate the density of neovascularization. (4) HE staining could be used for observe the infiltration of adipocytes and semi-quantitative analysis; (5) Masson staining could clearly reflect the deposition of collagen fiber, and the thickness of the fiber deposition area was measured and compared; (6) All observation indicators of the two groups were compared by quantitative and semi-quantitative methods, and there was no significant difference (P>0.05). ConclusionThe detection method adopted in this study could directly and accurately locate and quantify the indicators of local response, which basically met the requirements of "Guidelines for technical review of intraperitoneal hernia repair mesh in animal experiments" , and has a certain reference value for future relevant experiments.