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牛心包的高效去细胞化,同时保留细胞外基质并具有良好的生物相容性。

Efficient decellularization for bovine pericardium with extracellular matrix preservation and good biocompatibility.

作者信息

Li Ning, Li Yang, Gong Dejun, Xia Cuiping, Liu Xiaohong, Xu Zhiyun

机构信息

Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China.

Department of Cardiothoracic Surgery Lab, Changhai Hospital, Second Military Medical University, Shanghai, China.

出版信息

Interact Cardiovasc Thorac Surg. 2018 May 1;26(5):768-776. doi: 10.1093/icvts/ivx416.

Abstract

OBJECTIVES

In this study, we sought to explore an efficient decellularization protocol for bovine pericardia with better extracellular matrix preservation and good biocompatibility.

METHODS

Bovine pericardia were decellularized by sodium dodecyl sulphate (SDS), SDS + sodium deoxycholate (SD), Triton X-100 (TX), TX + SD (TS), freeze-thaw cycles + SDS + SD (FSS) and freeze-thaw cycles + TX + SD (FTS), respectively. Untreated pericardia were used as native control. Histological examination, residual cellular content analysis, biochemical and biomechanical evaluations and cytotoxicity assay were performed to investigate decellularization efficiency, xenoantigens removal, extracellular matrix preservation and biocompatibility. In vivo biocompatibility was evaluated using a subcutaneous implantation method in rats.

RESULTS

Among these protocols, FSS and FTS protocols were the most effective methods to remove both the DNA material and the galactose-α-1,3-galactose antigen. TX, TS and FTS bovine pericardia maintained the collagen content and had no cytotoxicity to human umbilical vein endothelial cells. The contents of elastin and glycosaminoglycan were lost to different degrees after decellularization, with the highest content of preservation with TX, followed by TS and FTS. Consistently, no significant difference was found between native bovine pericardia and TX, TS or FTS bovine pericardia. In vivo, FTS implants had minimal infiltration of macrophages and T-lymphocytes, with no histological evidence of peri-implant necrosis and calcification.

CONCLUSIONS

These results suggested that the FTS protocol showed optimal decellularization results with better extracellular matrix preservation and good biocompatibility. It may be a suitable protocol for producing a suitable scaffold for heart tissue engineering.

摘要

目的

在本研究中,我们试图探索一种高效的牛心包脱细胞方案,以更好地保存细胞外基质并具有良好的生物相容性。

方法

分别用十二烷基硫酸钠(SDS)、SDS + 脱氧胆酸钠(SD)、Triton X - 100(TX)、TX + SD(TS)、冻融循环 + SDS + SD(FSS)和冻融循环 + TX + SD(FTS)对牛心包进行脱细胞处理。未处理的心包用作天然对照。进行组织学检查、残余细胞含量分析、生化和生物力学评估以及细胞毒性测定,以研究脱细胞效率、异种抗原去除、细胞外基质保存和生物相容性。采用大鼠皮下植入法评估体内生物相容性。

结果

在这些方案中,FSS和FTS方案是去除DNA物质和半乳糖-α-1,3-半乳糖抗原最有效的方法。TX、TS和FTS处理的牛心包保持了胶原蛋白含量,对人脐静脉内皮细胞无细胞毒性。脱细胞后弹性蛋白和糖胺聚糖的含量不同程度丢失,TX处理的保存含量最高,其次是TS和FTS。同样,天然牛心包与TX、TS或FTS处理的牛心包之间未发现显著差异。在体内,FTS植入物的巨噬细胞和T淋巴细胞浸润最少,没有植入物周围坏死和钙化的组织学证据。

结论

这些结果表明,FTS方案显示出最佳的脱细胞效果,能更好地保存细胞外基质并具有良好的生物相容性。它可能是一种适合用于心脏组织工程制备合适支架的方案。

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