Degradation effect of diepoxide fixation on porcine endogenous retrovirus DNA in heart valves: molecular aspects.
OBJECTIVE
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.
Biological Heart Valve Laboratory, Foundation of Cardiac Surgery Development, Zabrze, Poland.
Aleksandra
A
Cyganek-Niemiec
Barbara
B
Strzalka-Mrozik
Lucyna
L
Pawlus-Lachecka
Jolanta
J
Wszolek
Jolanta
J
Adamska
Julia
J
Kudrjavtseva
Irina
I
Zhuravleva
Malgorzata
M
Kimsa
Hubert
H
Okla
Magdalena
M
Kimsa
Agnieszka
A
Gudek
Urszula
U
Mazurek
D000818:Animals; D001705:Bioprosthesis / Q000821:virology*; D004279:DNA, Viral / Q000187:drug effects* / Q000302:isolation & purification; D004853:Epoxy Resins / Q000494:pharmacology*; D005404:Fixatives / Q000494:pharmacology*; D005976:Glutaral / Q000494:pharmacology; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006350:Heart Valve Prosthesis; D019918:Heart Valve Prosthesis Implantation / Q000009:adverse effects / Q000295:instrumentation*; D006801:Humans; D011474:Prosthesis Design; D016459:Prosthesis-Related Infections / Q000517:prevention & control / Q000635:transmission / Q000821:virology; D060888:Real-Time Polymerase Chain Reaction; D012190:Retroviridae / Q000187:drug effects* / Q000235:genetics; D012192:Retroviridae Infections / Q000517:prevention & control / Q000635:transmission / Q000821:virology; D013552:Swine; D013696:Temperature; D013997:Time Factors; D016707:Tissue Fixation; D015047:Zoonoses / Q000635:transmission / Q000821:virology
D004279:DNA, Viral; D004853:Epoxy Resins; D005404:Fixatives; C035364:Quetol 651; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005976:Glutaral