Introduction

Bladder dysfunction has been considered as one of the most critical health conditions with no proper treatment. Current therapeutic approaches including enterocystoplasty have several limitations. Hence, biofabrication of cell-laden biological allografts using decellularized Goat urinary bladder scaffolds for organ reconstruction/regeneration was major objective of this study.

Materials and methods

An efficient method for decellularization of Goat urinary bladder (N = 3) was developed by perfusion of gradient change of detergents through ureter. The retention of organ architecture, extracellular matrix composition, mechanical properties and removal of cellular components was characterized using histological, cellular and molecular analysis. Further, mesenchymal stem cells (MSCs) from human umbilical cord blood (UCB) were used for preparing biological construct of decellularized urinary bladder (DUB) scaffolds to augment the urinary bladder reconstruction/regeneration.

Results

The decellularization method adopted in this study generated completely DUB scaffolds within 10 h at 100 mm Hg pressure and constant flow rate of 1 mL/min. The DUB scaffold retains organ architecture, ECM composition, and mechanical strength. No significant amount of residual nucleic acid was observed post-decellularization. Furthermore, MSCs derived from human UCB engrafted and proliferated well on DUB scaffolds in highly aligned manner under xeno-free condition.

Conclusion

Biofabricated humanized urinary bladder constructs provides xeno-free allografts for future application in augmenting urinary bladder reconstruction/regeneration with further development.

中文翻译：

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用于器官重建/再生的山羊膀胱支架的细胞负载同种异体移植物的生物制造

介绍

膀胱功能障碍被认为是最严重的健康状况之一，没有得到适当的治疗。目前包括肠囊成形术在内的治疗方法有几个局限性。因此，使用脱细胞山羊膀胱支架进行器官重建/再生的细胞负载生物同种异体移植物的生物制造是本研究的主要目标。

材料和方法

一种有效的山羊膀胱脱细胞方法 (N = 3) 是通过输尿管灌注梯度变化的洗涤剂开发的。使用组织学、细胞和分子分析表征器官结构的保留、细胞外基质组成、机械特性和细胞成分的去除。此外，来自人脐带血 (UCB) 的间充质干细胞 (MSC) 用于制备脱细胞膀胱 (DUB) 支架的生物构建体，以增强膀胱重建/再生。

结果

本研究中采用的脱细胞方法在 100 毫米汞柱压力和 1 毫升/分钟的恒定流速下在 10 小时内生成了完全 DUB 支架。DUB 支架保留了器官结构、ECM 组成和机械强度。脱细胞后未观察到显着量的残留核酸。此外，源自人类 UCB 的 MSC 在无异种条件下以高度对齐的方式在 DUB 支架上移植和增殖良好。

结论

生物制造的人性化膀胱结构提供了无异种同种异体移植物，用于未来在进一步发展中增强膀胱重建/再生的应用。