A dynamic distention protocol for whole-organ bladder decellularization: histological and biomechanical characterization of the acellular matrix
A dynamic distention protocol for whole-organ bladder decellularization: histological and biomechanical characterization of the acellular matrix
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DOI：
10.1002/term.1767
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发表时间：
2016-02-01
影响因子：
3.3
通讯作者：
Soncini, M.
中科院分区：
文献类型：
--
作者：
Consolo, F.;Brizzola, S.;Soncini, M.
提出了一种全层膀胱去细胞的物理-化学联合方案，该方案基于器官循环扩张，通过尿路反复输注/撤除脱细胞药物。动态脱细胞的目的是提高细胞去除效率，促进组织内层内洗涤剂的输送和细胞碎片的去除。使用温和的化学洗涤剂(低渗溶液和非离子洗涤剂)来限制对基质3D超微结构的不利影响。对脱细胞基质进行残留DNA和RNA的检查，以验证有效的细胞去除。组织学研究的重点是评估足够的结构和功能组件的保留，这些组件调节无细胞组织的生物力学行为。通过组织条的单轴拉伸加载试验和体外充盈膀胱测定法评价生物力学特性，以评估整个器官对生理类负荷状态的机械响应。根据我们的结果，在洗涤剂输注流量为5ml/min的情况下，动态脱细胞程序持续17h，比标准的静态脱细胞程序具有更高的脱细胞效率，导致残留的DNA含量
A combined physical-chemical protocol for whole full-thickness bladder decellularization is proposed, based on organ cyclic distention through repeated infusion/withdrawal of the decellularization agents through the urethra. The dynamic decellularization was intended to enhance cell removal efficiency, facilitating the delivery of detergents within the inner layers of the tissue and the removal of cell debris. The use of mild chemical detergents (hypotonic solution and non-ionic detergent) was employed to limit adverse effects upon matrix 3D ultrastructure. Inspection of the presence of residual DNA and RNA was carried out on decellularized matrices to verify effective cell removal. Histological investigation was focused on assessing the retention of adequate structural and functional components that regulate the biomechanical behaviour of the acellular tissue. Biomechanical properties were evaluated through uniaxial tensile loading tests of tissue strips and through ex vivo filling cystometry to evaluate the whole-organ mechanical response to a physiological-like loading state. According to our results, a dynamic decellularization protocol of 17 h duration with a 5 ml/min detergent infusion flow rate revealed higher DNA removal efficiency than standard static decellularization, resulting in residual DNA content