Central Laboratory for Stem Cell Research and Translational Medicine, Centre for Liver Research and Diagnostics, Deccan College of Medical Sciences, Kanchanbagh, Hyderabad 500058, Telangana, India.
Department of Biochemistry, Deccan College of Medical Sciences, Kanchanbagh, Hyderabad 500058, Telangana, India.
Tissue Cell. 2020 Dec;67:101443. doi: 10.1016/j.tice.2020.101443. Epub 2020 Sep 28.
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.
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.
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.
Biofabricated humanized urinary bladder constructs provides xeno-free allografts for future application in augmenting urinary bladder reconstruction/regeneration with further development.
膀胱功能障碍被认为是一种没有适当治疗方法的最严重的健康问题之一。目前的治疗方法包括肠膀胱成形术,但存在一些局限性。因此,本研究的主要目标是使用脱细胞山羊膀胱支架生物制造细胞负载的生物同种异体移植物,用于器官重建/再生。
通过输尿管灌注去污剂的梯度变化,开发了一种有效的山羊膀胱脱细胞方法(N = 3)。使用组织学、细胞和分子分析对器官结构、细胞外基质组成、机械性能和细胞成分的去除进行了表征。进一步,从人脐带血(UCB)中分离间充质干细胞(MSCs),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。
本研究采用的脱细胞方法在 100mm Hg 压力和 1ml/min 的恒定流速下,10 小时内可生成完全的 DUB 支架。DUB 支架保留了器官结构、ECM 组成和机械强度。脱细胞后未观察到明显的残留核酸。此外,在无异种条件下,从人 UCB 中分离出的 MSC 在 DUB 支架上很好地植入和增殖,并呈高度定向排列。
生物制造的人源化膀胱构建体为未来应用提供了无异种的同种异体移植物,用于增强膀胱重建/再生。