Efficient decellularization for bovine pericardium with extracellular matrix preservation and good biocompatibility
Efficient decellularization for bovine pericardium with extracellular matrix preservation and good biocompatibility
复制标题
牛心包的高效脱细胞，保留细胞外基质，生物相容性好
DOI：
10.1093/icvts/ivx416
复制
发表时间：
2018-05-01
影响因子：
--
通讯作者：
Xu, Zhiyun
中科院分区：
文献类型：
--
作者：
Li, Ning;Li, Yang;Xu, Zhiyun
目的：在本研究中，我们试图探索一种具有更好的细胞外基质保存和良好生物相容性的牛心包高效脱细胞方案。方法：分别采用十二烷基硫酸钠（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。未治疗的心包作为自然对照。通过组织学检查、残留细胞含量分析、生物化学和生物力学评价以及细胞毒性试验来研究脱细胞效率、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价其体内生物相容性。结果：FSS和FTS是去除DNA材料和半乳糖- α -1,3-半乳糖抗原最有效的方法。TX、TS和FTS牛心包维持胶原蛋白含量，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含量均有不同程度的损失，其中TX保存含量最高，TS和FTS次之。一致地，在原生牛心包和TX、TS或FTS牛心包之间没有发现显著差异。在体内，FTS种植体中巨噬细胞和t淋巴细胞的浸润很少，未见种植体周围坏死和钙化的组织学证据。结论：FTS脱细胞效果最佳，细胞外基质保存效果好，生物相容性好。这可能是制造适合心脏组织工程的支架的一种合适的方案。
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-alpha-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.