网膜包裹组织工程移植物构建复层结构的膀胱补片  

Bladder patch construction with multilayer structure by tissue engineering graft wrapped up in omentum

作　　者：李欣慧 俞远东 喻雄杰 吴光 Li Xinhui;Yu Yuandong;Yu Xiongjie;Wu Guang(Department of Cancer Center,The Affiliated People’s Hospital of Hubei University of Medicine)

机构地区：[1]湖北医药学院附属人民医院肿瘤中心,十堰442000

出　　处：《重庆医科大学学报》2021年第4期454-459,共6页Journal of Chongqing Medical University

摘　　要：目的:利用网膜包裹膀胱脱细胞基质(bladder acellular matrix,BAM)复合多种细胞的组织工程移植物构建复层结构且血管化丰富的组织工程膀胱补片。方法:将体外扩增培养的兔骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)、膀胱平滑肌细胞、膀胱上皮细胞分别以1.0×106个/mL先后种植于制备的BAM上,培养10 d后制作成膀胱上皮细胞-平滑肌细胞-BMSCs-BAM组织工程膀胱补片;实验组将组织工程膀胱补片包裹在硅胶导管上植入兔网膜间培养,对照组单纯将BAM补片包裹在硅胶导管上植入兔网膜间培养,2组分别于4周和8周后取材行免疫组化,采用CKAE1/AE3抗体检测上皮层,α-SMA抗体检测平滑肌层及CD31抗体检测血管生成情况。结果:4周后,实验组膀胱补片有上皮层和平滑肌层形成,且内部有明显的新生血管,CKAE1/AE3、α-SMA及CD31表达阳性;8周后,实验组上皮层及平滑肌层增厚,血管增多增粗。对照组BAM补片仅有微小血管形成,无上皮层和平滑肌层形成。结论:将膀胱上皮细胞、平滑肌细胞和BMSCs与BAM复合后行体内网膜包裹培养,可构建复层结构且血管化丰富的组织工程膀胱补片。Objective:To construct a tissue-engineered bladder patch with multilayer structure and abundant vascularization on the basis of bladder acellular matrix(BAM)combined with various cells wrapped up in omentum.Methods:Bone marrow mesenchymal stem cells(BMSCs),bladder epithelial cells and bladder smooth muscle cells of rabbits were seeded on the surface of BAM at 1.0×106 cells/mL,respectively.After 10 days of cultivation,bladder epithelial cells-smooth muscle cells-BMSCs-BAM tissue engineered bladder patch was prepared.In the experimental group,tissue engineered bladder patch were wrapped on silica gel catheter and implanted into rabbit omentum for culture.In the control group,BAM patch was wrapped on silica gel catheter and implanted into rabbit omentum for culture.Samples in two groups were taken for immunohistochemistry after 4 weeks and 8 weeks respectively.The epithelial layer was detected by CKAE1/AE3 antibody,the smooth muscle layer was detected byα-SMA antibody and the angiogenesis was detected by CD31 antibody.Results:After 4 weeks,epithelial layer and smooth muscle layer were formed in the experimental group,and obvious neovascularization were found in the bladder patch and CKAE1/AE3,α-SMA and CD31 were expressed positively.After8 weeks,the epithelial layer and smooth muscle layer in the experimental group became thicker,and the blood vessels became denser and thicker.However,the BAM patch in the control group had only minute angiogenesis,without epithelial layer and smooth muscle layer.Conclusion:Tissue engineered bladder patch with multilayer structure and abundant vascularization can be constructed by BAM combined with bladder epithelial cells,smooth muscle cells and BMSCs and wrapped up in omentum.

关 键 词：膀胱上皮细胞 膀胱平滑肌细胞 骨髓间充质干细胞 网膜包裹 膀胱补片 

分 类 号：R318.5[医药卫生—生物医学工程]