兔脱细胞尿道基质制备的可行性

Feasibility of the preparation of acellular urethral matrix graft

目的:采用曲拉松与氨水混合液萃取制备脱细胞尿道基质材料,探讨此方法的可行性。方法:实验于2002-12/2003-03在中山大学附属第二医院整形外科完成。选用新西兰雄性大白兔4只,取标本前2天起每天肌注庆大霉素1万单位。将4只新西兰大白兔采用空气栓塞处死后,在无菌条件下取完整尿道组织,用曲拉松与氨水混合液萃取制备脱细胞尿道基质材料,分别于脱细胞处理3,7,11d后收集标本进行物理性状观察、组织学观察,找到脱细胞效果最佳的时间段。结果:实验纳入大白兔4只,全部进入结果分析。①脱细胞处理前后尿道基质的物理性状观察:经脱细胞处理后的尿道基质,其外观体积与脱细胞处理前相差不大,但颜色变为透白色,柔软,富有弹性及韧性。②脱细胞处理后不同时间尿道基质组织学观察:与正常尿道组织相比,脱细胞处理3d后组织中蓝染核物质大量减少,局部核物质聚集,有细胞碎屑残留,弹力纤维排列规整,管腔光滑;脱细胞处理7d后组织中蓝染核物质进一步减少,视野中剩余核物质呈线形或弧形排列,仍有细胞碎屑残留,弹力纤维排列整齐,但空隙较正常加大,管腔光滑;脱细胞处理11d后苏木精-伊红染色蓝染核物质和细胞碎屑已不存在,弹力纤维排列仍规整,空隙较前述增大,管腔光滑,脱细胞处理成功。结论:采用曲拉松及氨水混合液经脱细胞处理11d后,尿道基质组织材料中未见细胞存在,由排列规则的胶原及弹力纤维组成,管腔光滑,证明脱细胞尿道基质制备成功。

AIM： To evaluatethe feasibility of the preparation of acellular urethral matrix graft treated by solution of Triton-X100 and NH3·H2O. METHODS： The experiment was completed in the Department of Plastic and Reconstructive Surgery, the Second Affiliated Hospital of Sun Yat-sen University from November 2002 to March 2003. Four New Zealand male white rabbits were used, all the rabbits were injected with Gentamycin 10 000 units＇per day from 2 days before being taken the urethras. The 4 rabbits were executed by air embolism, the complete urethras were excised under sterile conditions. The urethras were treated by solution of Triton-X100 and NH3·H2O to remove cells, and then the physical and histological characters of the specimen were observed at 3, 7 and 11 days after extraction to confirm the right time of successful cellular elimination. RESULTS： All the 4 rabbits entered the analysis of result. ① Observation of the physical characters of the urethra matrix after removing cells： Its physical volume was not greatly different from that before removing cells processing, but the color change into deeply white, soft, full of the flexibility and tenacity. ② Observation the histological characters of the urethra matrix after removing cells： Compared with the normal urethral tissue, removing cells for 3 days, the blue dyed materials decreased obviously, parts of nuclear materials came together, broken cell bits remained, the elasticity fiber arranged tidily, the lumen surface was smooth; Removing cells for 7 days, the blue dyed materials decreased further, in the visual field the surplus nuclear material presented the line form or arc-shaped alignment, the cell broken bits remained, the elasticity fiber arranged tidy, but the crevices were much larger, the lumen surface was smooth; Removing cells for 11 days, the blue dyed materials and the broken cell bits were already nonexistent, the elasticity fiber still arranged tidy, the crevices were more larger, the lumen surface was smooth, removed cells successfully. CONCLUSION： After cellular extracting in solution of Triton-X100 and NH3·H2O. for 11 days, the cells are nonexistent, the matrix is make up of collagen and elastin that arranging tidily. Acellular urethral matrix graft can be prepared successfully by cellular extracting.