Bladder reconstruction using autologous smooth muscle cell sheets grafted on a pre-vascularized capsule
Bladder reconstruction using autologous smooth muscle cell sheets grafted on a pre-vascularized capsule
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使用移植到预血管化胶囊上的自体平滑肌细胞片进行膀胱重建
DOI：
10.7150/thno.47006
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发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Chen, Fang
中科院分区：
文献类型：
--
作者：
Guo, Hai-Lin;Peng, Xu-Feng;Chen, Fang
理论基础：构建功能血管化的三维组织一直是组织工程领域的长期目标。应用组织扩张器胶囊作为诱导血管床，在兔膀胱重建模型上预制功能性血管蒂肌组织瓣进行膀胱重建的效果。方法：在腹股沟置入皮肤组织扩张器，诱导带血管的囊袋形成。取血管内皮祖细胞与血管内皮细胞共培养，形成血管预制的血管内皮细胞片。然后，每隔2天将三层细胞片移植到血管化的包膜组织上，以预制功能性的血管化平滑肌组织瓣。分别采用六层细胞片(单纯片状)、囊膜瓣(单纯囊膜)和带血管的囊膜(片状+囊膜)修复膀胱肌肌壁缺损。动物植入后随访3个月，连续移位膀胱。结果：片剂加胶囊组3个月内膀胱容量和顺应性均保持不变。组织浴刺激显示三组大鼠对卡巴胆碱和氯化钾的收缩反应有显著差异(p<；0.05)。组织学上，胶囊组1个月时炎症明显，片剂组3个月时可见纤维化。各时间点单纯胶囊组和片剂+胶囊组的血管密度均显著高于片剂组(p<；0.05)。三组间的平滑肌含量比较，差异有统计学意义(p<；0.05)。结论：该胶囊可作为血管蒂肌组织瓣预制的诱导血管床。预制的功能性血管蒂肌组织瓣有可能用于可靠的膀胱重建，并可能为三维组织工程中的血管化创造新的机会。
Rationale: Construction of functional vascularized three-dimensional tissues has been a longstanding objective in the field of tissue engineering. The efficacy of using a tissue expander capsule as an induced vascular bed to prefabricate functional vascularized smooth muscle tissue flaps for bladder reconstruction in a rabbit model was tested. Methods: Skin tissue expanders were inserted into the groin to induce vascularized capsule pouch formation. Smooth muscle cells and endothelial progenitor cells were harvested and cocultured to form pre-vascularized smooth muscle cell sheet. Then repeated transplantation of triple-layer cell sheet grafts onto the vascularized capsular tissue was performed at 2-day intervals to prefabricate functional vascularized smooth muscle tissue flaps. Bladder muscular wall defects were created and repaired by six-layer cell sheet graft (sheet only), capsule flap (capsule only) and vascularized capsule prelaminated with smooth muscle cell sheet (sheet plus capsule). The animals were followed for 3 months after implantation and their bladders were explanted serially. Results: Bladder capacity and compliance were maintained in sheet plus capsule group throughout the 3 months. Tissue bath stimulation demonstrated that contractile responses to carbachol and KCl among the three groups revealed a significant difference (p < 0.05). Histologically, inflammation was evident in the capsule only group at 1 month and fibrosis was observed in sheet only group at 3 months. The vessel density in capsule only and sheet plus capsule group were significantly higher than in the sheet only group at each time point (p < 0.05). Comparison of the smooth muscle content among the three groups revealed a significant difference (p < 0.05). Conclusion: These results proved that the capsule may serve as an induced vascular bed for vascularized smooth muscle tissue flap prefabrication. The prefabricated functional vascularized smooth muscle tissue flap has the potential for reliable bladder reconstruction and may create new opportunities for vascularization in 3-D tissue engineering.