Background For muscle invasive bladder cancer, radical cystectomy is the most effective treatment now and urinary diversion is often necessary. The use of intestinal tissue for urinary diversion is frequently associat...Background For muscle invasive bladder cancer, radical cystectomy is the most effective treatment now and urinary diversion is often necessary. The use of intestinal tissue for urinary diversion is frequently associated with complications. In this study, we aimed to make a tissue-engineered conduit （TEC） using bladder epithelial cells and bladder acellular matrix （BAM） for urinary diversion in rabbits. Methods Bladder epithelial cells of rabbit were cultivated and expanded in vitro, then seeded on BAM, and cultured for 7 days. Then cell-seeded graft was used to make TEC. In the experimental group, most of bladder of the rabbit was removed while bladder trigone was retained. The proximal end of TEC was anastomosed with bladder trigone and the distal end was anastomosed with the abdominal stoma. In the control group, TEC was made using unseeded BAM. Haematoxylin and eosin staining was conducted, respectively, at 1, 2, 4, and 8 weeks postoperatively. Immunohistochemistry was performed 8 weeks postoperatively. Intravenous urography, retrograde pyelography, and cystoscopy of TEC were made at 12 weeks postoperatively. Results All animals were alive in the experimental group. Haematoxylin and eosin staining showed epithelial coverage in TEC. Immunohistochemistry showed anti-cytokeratin AEI/AE3 antibody and anti-ZO1 antibody positive, confirming there were mature and functional epithelial cells on the lumen of TEC. Retrograde pyelography and intravenous urography showed that TEC developed well and that there was no obstruction. In the control group, four rabbits were dead within 2 weeks and scar formation, atresia, and severe hydronephrosis were found. Conclusions We successfully made TEC using BAM and bladder epithelial cells for urinary diversion in rabbits. The lumen of this new TEC covered mature epithelial cells and could prevent urinary extravasation.展开更多
Objective:To study the biocompatibility and neovascularization of the PLGA nanospheres wrapped with vascular endothelial growth factor(VEGF).which can improve bladder acellular matrix graft(BAMG) with local continuous...Objective:To study the biocompatibility and neovascularization of the PLGA nanospheres wrapped with vascular endothelial growth factor(VEGF).which can improve bladder acellular matrix graft(BAMG) with local continuous release of VEGF.Methods:A total of 18 rabbit model (length of stenosis:3cm) with anterior urethral stricture were used as experimental animals and divided into three groups.Group A as the control group:Simple BAMG scaffold materials for urethral reconstruction.Group B as the blank group:PLGA microspheres modified BAMG for urethral reconstruction.Group C:PLGA conjugated with VEGF and modified BAMG for the urethral reconstruction.All rabbits underwent urethral angiography after 7 days,15 days,1 month and 3 months after the operation,and one rabbit in each group was sacrificed to be prepared for the organization histologic examination,HE staining,masson staining,CD31,34 and a-SAM immunohistochemical detection in the repaired sites.Results:In group A,significant urethral restenosis occurred in two rabbits after 15 days of the operation,HE and masson staining showed a lot of collagen arranged in the repaired sites,and there were a large number of inflammatory cell infiltration,and there were also CD31,34 in the repaired sites.a-SAM microvascular tag count showed a small amount of microvascular;Croup B showed anastomotic restenosis,HE and masoon staining showed inflammatory cell infiltration and collagen deposition;Group C:urethrography showed lumen patency.There were a small amount of inflammatory cell infiltration after 7 and 15 days after the operation,and there were also CD31,34 in the repaired sites.The a-SAM microvascular tag count showed many microvascular.And the difference was significant.Conclusions:Anterior urethral reconstruction with sustained-release of VEGF by PLGA nanospheres modified BAMG stents can reduce postoperative restenosis.It can also reduce collagen deposition and scar formation,promote angiogenesis of the repair tissue;therefore it in valuable in the tissue-engineered urethral reconstruction.展开更多
基金This work was supported by the National Science Foundation for Young Scientists of China （No. 81001019） and the Fundamental Research Funds for the Central Universities （No. 201130202020006）.
文摘Background For muscle invasive bladder cancer, radical cystectomy is the most effective treatment now and urinary diversion is often necessary. The use of intestinal tissue for urinary diversion is frequently associated with complications. In this study, we aimed to make a tissue-engineered conduit （TEC） using bladder epithelial cells and bladder acellular matrix （BAM） for urinary diversion in rabbits. Methods Bladder epithelial cells of rabbit were cultivated and expanded in vitro, then seeded on BAM, and cultured for 7 days. Then cell-seeded graft was used to make TEC. In the experimental group, most of bladder of the rabbit was removed while bladder trigone was retained. The proximal end of TEC was anastomosed with bladder trigone and the distal end was anastomosed with the abdominal stoma. In the control group, TEC was made using unseeded BAM. Haematoxylin and eosin staining was conducted, respectively, at 1, 2, 4, and 8 weeks postoperatively. Immunohistochemistry was performed 8 weeks postoperatively. Intravenous urography, retrograde pyelography, and cystoscopy of TEC were made at 12 weeks postoperatively. Results All animals were alive in the experimental group. Haematoxylin and eosin staining showed epithelial coverage in TEC. Immunohistochemistry showed anti-cytokeratin AEI/AE3 antibody and anti-ZO1 antibody positive, confirming there were mature and functional epithelial cells on the lumen of TEC. Retrograde pyelography and intravenous urography showed that TEC developed well and that there was no obstruction. In the control group, four rabbits were dead within 2 weeks and scar formation, atresia, and severe hydronephrosis were found. Conclusions We successfully made TEC using BAM and bladder epithelial cells for urinary diversion in rabbits. The lumen of this new TEC covered mature epithelial cells and could prevent urinary extravasation.
文摘Objective:To study the biocompatibility and neovascularization of the PLGA nanospheres wrapped with vascular endothelial growth factor(VEGF).which can improve bladder acellular matrix graft(BAMG) with local continuous release of VEGF.Methods:A total of 18 rabbit model (length of stenosis:3cm) with anterior urethral stricture were used as experimental animals and divided into three groups.Group A as the control group:Simple BAMG scaffold materials for urethral reconstruction.Group B as the blank group:PLGA microspheres modified BAMG for urethral reconstruction.Group C:PLGA conjugated with VEGF and modified BAMG for the urethral reconstruction.All rabbits underwent urethral angiography after 7 days,15 days,1 month and 3 months after the operation,and one rabbit in each group was sacrificed to be prepared for the organization histologic examination,HE staining,masson staining,CD31,34 and a-SAM immunohistochemical detection in the repaired sites.Results:In group A,significant urethral restenosis occurred in two rabbits after 15 days of the operation,HE and masson staining showed a lot of collagen arranged in the repaired sites,and there were a large number of inflammatory cell infiltration,and there were also CD31,34 in the repaired sites.a-SAM microvascular tag count showed a small amount of microvascular;Croup B showed anastomotic restenosis,HE and masoon staining showed inflammatory cell infiltration and collagen deposition;Group C:urethrography showed lumen patency.There were a small amount of inflammatory cell infiltration after 7 and 15 days after the operation,and there were also CD31,34 in the repaired sites.The a-SAM microvascular tag count showed many microvascular.And the difference was significant.Conclusions:Anterior urethral reconstruction with sustained-release of VEGF by PLGA nanospheres modified BAMG stents can reduce postoperative restenosis.It can also reduce collagen deposition and scar formation,promote angiogenesis of the repair tissue;therefore it in valuable in the tissue-engineered urethral reconstruction.
