缓释bFGF胶原膜修复猪胆管缺损的实验研究  

EFFECT OF SUSTAINED-RELEASE BASIC FIBROBLAST GROWTH FACTOR ON HEALING OF BILE DUCT DEFECT IN PIGS

作　　者：陶亮[1] 李强[1] 任昊桢[1] 戴建武[2] 丁义涛[1] 

机构地区：[1]南京大学医学院附属鼓楼医院肝胆外科,南京210008 [2]中国科学院遗传与发育生物学研究所

出　　处：《中国修复重建外科杂志》2013年第2期212-218,共7页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：南京市医学科技发展项目(YKK11087)~~

摘　　要：目的探讨采用缓释bFGF胶原膜修复猪胆管缺损的效果。方法中华实验用猪28只,雌雄不限,体重15～30 kg;根据处理方法不同,将其随机分为3组,其中实验组12只,空白对照组12只,正常对照组4只。正常对照组不作处理;实验组和空白对照组制备胆管壁缺损模型(长2 cm,宽1/3～2/3胆管周径)后,分别采用2.0 cm×1.0 cm×0.5 mm缓释bFGF胶原膜和空白胶原膜缝合修复。术后观察实验动物存活情况,并于术后1、2、3个月检测肝功能变化,处死动物后行大体观察;取胆管组织经HE及免疫组织化学染色,行微血管密度(microvessel density,MVD)及黏膜下腺体计数;术后3个月行经胆囊胆管胆道造影观察。结果术后实验动物均存活至实验完成。术后1个月实验组及空白对照组腹腔粘连均较严重,随着时间延长,实验组粘连较空白对照组明显缓解。肝功能检测示,除ALP在术后2、3个月实验组及空白对照组均较正常对照组显著下降(P<0.05)外,其余各指标,如谷丙转氨酶、总胆红素和白蛋白术后各时间点组间比较,差异均无统计学意义(P>0.05)。组织学及免疫组织化学染色观察示,实验组胆管腺体再生和上皮再生速度快于空白对照组,2个月即完成胆管上皮覆盖,3个月与正常对照组相似,相应组织水肿和炎性浸润均较空白对照组轻。术后1个月实验组黏膜下腺体计数及MVD值均显著高于两个对照组(P<0.05),2个月后下降并稳定于正常对照组水平;空白对照组随时间延长呈上升趋势,术后1、2个月显著低于实验组和正常对照组(P<0.05),3个月时达到并稳定于正常对照组水平。相关性分析表明,正常对照组、实验组和空白对照组术后各时间点黏膜下腺体计数与MVD之间均成正相关(P<0.01)。术后3个月造影显示,实验组、空白对照组均无胆管扩张及结石形成。结论缓释bFGF胶原膜可通过促进新生血管再生等方式促进猪胆管缺损愈合。Objective To investigate the effects of sustained-release basic fibroblast growth factor （bFGF） on healing of bile duct defect. Methods A model of bile duct wall defect （2 cm in length and 1/3-2/3 of the bile duct circumference in width） was made in 24 pigs （male or female, weighing 15-30 kg）, and then defect was repaired with sustained-release bFGF collagen membrane （2.0 cm× 1.0 cm× 0.5 cm in size） in the experimental group （n=12） or with collagen membrane （2.0 cm × 1.0 cm × 0.5 cm in size） alone in the blank control group （n=12）. Another 4 healthy pigs were used to obtain normal bile duct as normal control group. The survival condition of pigs was observed after operation; at 1, 2, and 3 months after operation, the blood sampling was collected to test the changes of liver function, and the bile duct specimens were harvested to count the microvessel density （MVD） and submucosal gland by HE staining and immunohistochemistry staining; and at 3 months after operation, cholangiography examination was done. Results All the animals survived to completion of the experiment. Intra-abdominal adhesion was serious in the experimental and blank control groups at 1 week after operation, but the adhesion was markedly improved in the experimental group when compared with the blank control group with time passing. The liver function test showed that alkaline phosphatase in the experimental group was significantly lower than that in the blank control group at 2 and 3 months （P 〈 0.05）, but no significant difference in aspartate aminortransferase, total bilirubin, and albumin was found among 3 groups （P 〉 0.05）. The histology and immunohistochemistry staining observations showed that the regeneration rates of submucosal glands and epithelium in the experimental group were faster than those in the blank control group; defect was covered with the epithelium at 2 months, and the structure was similar to that of normal control group at 3 months; and the edema and inflammation infiltrati

关 键 词：组织工程 胆管缺损 缓释bFGF 胶原 微血管密度 猪 

分 类 号：R657.4[医药卫生—外科学]