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双环氧固定对心脏瓣膜中猪内源性逆转录病毒DNA的降解作用:分子层面

Degradation effect of diepoxide fixation on porcine endogenous retrovirus DNA in heart valves: molecular aspects.

作者信息

Cyganek-Niemiec Aleksandra, Strzalka-Mrozik Barbara, Pawlus-Lachecka Lucyna, Wszolek Jolanta, Adamska Jolanta, Kudrjavtseva Julia, Zhuravleva Irina, Kimsa Malgorzata, Okla Hubert, Kimsa Magdalena, Gudek Agnieszka, Mazurek Urszula

机构信息

Biological Heart Valve Laboratory, Foundation of Cardiac Surgery Development, Zabrze, Poland.

出版信息

Int J Artif Organs. 2012 Jan;35(1):25-33. doi: 10.5301/ijao.5000071.

Abstract

PURPOSE

Xenotransplantations of porcine cells, tissues, and organs involve a risk of zoonotic viral infections in recipients, including by porcine endogenous retroviruses (PERVs), which are embedded the genome of all pigs. An appropriate preparation of porcine heart valves for transplantation can prevent retroviral infection. Therefore, the present study focuses on the effect of epoxy compounds and glutaraldehyde on the PERV presence in porcine heart valves prepared for clinical use.

METHODS

Porcine aortic heart valves were fixed with ethylene glycol diglycidyl ether (EDGE) at 5 °C and 25 °C as well as with glutaraldehyde (GA) for 4 weeks. Salting out was used to isolate genomic DNA from native as well as EDGE- and GA-fixed fragments of valves every week. Quantification of PERV-A, PERV-B, and PERV-C DNA was performed by real-time quantitative polymerase chain reaction (QPCR).

RESULTS

All subtypes of PERVs were detected in native porcine aortic heart valves. The reduction of the PERV-A, PERV-B, and PERV-C DNA copy numbers was observed in the heart valves which were EDGE-fixed at both temperatures, and in GA-fixed ones in the following weeks. After 7 and 14 days of EDGE cross-linking, significant differences between the investigated temperatures were found for the number of PERV-A and PERV-B copies. PERV DNA was completely degraded within the first week of EDGE fixation at 25 °C.

CONCLUSIONS

EDGE fixation induces complete PERV genetic material degradation in porcine aortic heart valves. This suggests that epoxy compounds may be alternatively used in the preparation of bioprosthetic heart valves in future.

摘要

目的

猪细胞、组织和器官的异种移植会使接受者面临人畜共患病毒感染的风险,包括感染猪内源性逆转录病毒(PERV),这种病毒嵌入在所有猪的基因组中。对用于移植的猪心脏瓣膜进行适当处理可预防逆转录病毒感染。因此,本研究聚焦于环氧化合物和戊二醛对临床用猪心脏瓣膜中PERV存在情况的影响。

方法

将猪主动脉心脏瓣膜分别在5℃和25℃用乙二醇二缩水甘油醚(EDGE)以及用戊二醛(GA)固定4周。每周采用盐析法从天然瓣膜以及EDGE和GA固定的瓣膜片段中分离基因组DNA。通过实时定量聚合酶链反应(QPCR)对PERV - A、PERV - B和PERV - C DNA进行定量分析。

结果

在天然猪主动脉心脏瓣膜中检测到了PERV的所有亚型。在两个温度下用EDGE固定的心脏瓣膜以及在接下来几周用GA固定的心脏瓣膜中,均观察到PERV - A、PERV - B和PERV - C DNA拷贝数减少。在EDGE交联7天和14天后,在所研究的温度之间,PERV - A和PERV - B拷贝数存在显著差异。在25℃下用EDGE固定的第一周内,PERV DNA完全降解。

结论

EDGE固定可导致猪主动脉心脏瓣膜中PERV遗传物质完全降解。这表明环氧化合物未来可能可替代地用于生物人工心脏瓣膜的制备。

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