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曲拉通X-100与胰凝乳蛋白酶结合:一种制备去细胞猪颈动脉的更具前景的方案。

Triton X-100 combines with chymotrypsin: A more promising protocol to prepare decellularized porcine carotid arteries.

作者信息

Wang Fei, Zhang Jian, Wang Rong, Gu Yongquan, Li Jianxin, Wang Cong

机构信息

Department of Vascular Surgery, Xuan Wu Hospital, Capital Medical University, Beijing, P.R. China.

Institute of Vascular Surgery, Capital Medical University, Beijing, P.R. China.

出版信息

Biomed Mater Eng. 2017;28(5):531-543. doi: 10.3233/BME-171694.

Abstract

BACKGROUND

Morbidity and mortality of cardiovascular diseases are increasing in recent years. To solve these problems, vascular transplantation has become a common approach. Decellularization has been a hot spot of tissue engineering to prepare vessel substitutes for vascular transplantation. However, there is no established canonical protocol for decellularization thus far.

OBJECTIVE

To further understand the decellularization effect of decellularization protocols and the causal relationship between decellularization and mechanical properties.

METHODS

Three decellularization protocols including two chemical protocols based on SDS and Trypsin respectively and a combination of Triton X-100 with chymotrypsin were adopted to obtain decellularized porcine carotid arteries in our study. After decellularization, histological analysis, scanning electron microscopy and mechanical tests were performed to evaluate their efficiency on removing of cellular components, retention of extracellular matrix and influence on mechanical properties.

RESULTS

All these decellularization protocols used in our study were efficient to remove cellular components. However, SDS and trypsin performed more disruptive effect on ECM structure and mechanical properties of native arteries while Triton X-100 combines with chymotrypsin had no significant disruptive effect.

CONCLUSIONS

Compared with decellularization protocols based on SDS and trypsin, Triton X-100 combines with chymotrypsin used in our study may be a more promising protocol to prepare decellularized porcine carotid arteries for vascular tissue engineering applications.

摘要

背景

近年来,心血管疾病的发病率和死亡率不断上升。为了解决这些问题,血管移植已成为一种常用方法。去细胞化已成为组织工程领域的研究热点,用于制备血管移植的血管替代物。然而,迄今为止,尚未建立标准化的去细胞化方案。

目的

进一步了解去细胞化方案的去细胞化效果以及去细胞化与力学性能之间的因果关系。

方法

本研究采用三种去细胞化方案,分别是基于十二烷基硫酸钠(SDS)和胰蛋白酶的两种化学方案以及曲拉通X-100与胰凝乳蛋白酶的联合方案,以获取去细胞化的猪颈动脉。去细胞化后,进行组织学分析、扫描电子显微镜检查和力学测试,以评估它们在去除细胞成分、保留细胞外基质以及对力学性能的影响方面的效率。

结果

本研究中使用的所有这些去细胞化方案都能有效去除细胞成分。然而,SDS和胰蛋白酶对天然动脉的细胞外基质结构和力学性能具有更大的破坏作用,而曲拉通X-100与胰凝乳蛋白酶联合使用则没有显著的破坏作用。

结论

与基于SDS和胰蛋白酶的去细胞化方案相比,本研究中使用的曲拉通X-100与胰凝乳蛋白酶联合方案可能是一种更有前景的方案,用于制备用于血管组织工程应用的去细胞化猪颈动脉。

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