异源性膀胱脱细胞基质移植修复兔膀胱缺损的局部和全身安全性评价  

LOCAL AND SYSTEMIC SAFETY EVALUATION OF REGENERATED RABBIT BLADDER REPAIRED WITH XENOGENEOUS BLADDER ACELLULAR MATRIX

作　　者：周六化[1] 杨斌[1,2,3] 王鹏基[1] 夏佳东[1] 章宜芬[4] 陈赟[1] 戴玉田[1] 

机构地区：[1]南京大学医学院附属鼓楼医院泌尿男科,南京210008 [2]上海市第十人民医院 [3]暨同济大学附属第十人民医院泌尿外科 [4]南京大学医学院附属鼓楼医院病理科,南京210008

出　　处：《中国修复重建外科杂志》2013年第8期935-944,共10页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(31100702、81170563);高等学校博士学科点专项科研基金资助项目(20110072120054)~~

摘　　要：目的采用异源性膀胱脱细胞基质(bladder acellular matrix,BAM)移植修复兔膀胱缺损,检测组织再生和机体的组织反应情况,评价其生物安全性。方法将完整新鲜猪膀胱通过一系列物理、化学和酶消化法进行脱细胞处理,得到猪BAM,通过HE染色和扫描电镜观察其脱细胞效果和超微结构。将18只新西兰大白兔(体重2.5～3.0 kg)随机分为实验组(n=12)和对照组(n=6),行膀胱部分(约30%)切除术,实验组采用与切除面积等大的猪BAM替代修补,对照组直接行膀胱切口原位缝合。术后观察动物存活情况,分别于术后15 d,1、2、3、6个月行血常规及肾功能、电解质检测。术后1、2、3、6个月行尿流动力学检测最大膀胱容量、排尿点膀胱内压力以及膀胱顺应性;然后处死动物,大体观察膀胱再生情况,并取再生膀胱组织行HE染色及免疫组织化学染色;同时取周围脏器及淋巴组织,经大体观察及HE染色观察有无异常变化。结果制备的猪BAM完全去除了细胞成分,扫描电镜观察呈多孔样结构。实验动物均存活至取材各时间点,血常规检测示术后15 d两组白细胞计数升高,之后逐渐恢复至正常水平,两组间差异无统计学意义(P>0.05)。肾功能、电解质检测提示两组间差异无统计学意义(P>0.05),其中肌酐有上升趋势,但至术后6个月时仍维持在正常范围内。术后实验组最大膀胱容量及膀胱顺应性与对照组相比明显恢复,术后3、6个月两组比较差异有统计学意义(P<0.05);两组间各时间点排尿点膀胱内压力比较差异均无统计学意义(P>0.05)。组织学观察可见两组中均有尿路上皮层、平滑肌组织和血管的再生。两组术后1个月均可见慢性炎性细胞浸润,随后慢性炎性细胞明显减少(P<0.05),直至基本消退;术后3、6个月,两组间慢性炎性细胞浸润程度评分比较差异无统计学意义(P>0.05)。随着时间推移,两组α-平滑肌肌动蛋白(α-smooth muscleObjective To evaluate tissue regeneration,body reaction,and biological safety of xenogeneous bladder acellular matrix（BAM） that can be used to repair rabbit bladder. Methods Porcine BAM was prepared through physical,chemical,and enzymatic methods,and the effects of acellularization and the structure were observed with HE staining and scanning electron microscope（SEM）.Eighteen New Zealand white rabbits（weighing,2.5-3.0 kg） undergoing partial cystectomy were randomly divided into 2 groups.After partial（about 30%） cystectomy,the porcine BAM was used to replace partial rabbit bladder in the experimental group（n=12）,and the incision was directly sutured as control group（n=6）.The survival condition of animals was observed after operation.At 15 days,1,2,3,and 6 months after operation,the blood routine,renal function,and electrolyte were tested by collecting the blood samples.At 1,2,3,and 6 months after operation,maximum bladder capacity,bladder leak point pressure,and bladder compliance were measured through urodynamic studies.Then gross observation was performed for regeneration of bladder,and the specimens of the bladder were harvested for HE staining and immunohistochemical staining.The surrounding organs and local lymphoid tissues were harvested for gross observation and HE staining. Results Cell components were completely removed in the porcine BAM,showing three-dimensional porous structure under SEM.All the animals survived during the experiment.At 15 days after operation,white blood cell count increased,and then returned to normal level in 2 groups,showing no significant difference between 2 groups（P 0.05）.The tests of renal function and electrolyte suggested no significant difference between 2 groups（P 0.05）.The level of serum creatinine showed a tendency of increase,but it remained within normal range at 6 months after operation.The maximum bladder capacity and compliance in experimental group were significantly higher than those in control group at 3 and 6 months after operation

关 键 词：膀胱再生 膀胱脱细胞基质 异种移植 安全性评价 猪 兔 

分 类 号：R699.5[医药卫生—泌尿科学]