PMID- 31692126
OWN - NLM
STAT- MEDLINE
DCOM- 20210608
LR  - 20210608
IS  - 1399-3089 (Electronic)
IS  - 0908-665X (Linking)
VI  - 27
IP  - 2
DP  - 2020 Mar
TI  - Decellularized pig pulmonary heart valves-Depletion of nucleic acids measured by 
      proviral PERV pol.
PG  - e12565
LID - 10.1111/xen.12565 [doi]
AB  - BACKGROUND: Decellularized human pulmonary heart valve (dhHV) scaffolds have been 
      shown to be the gold standard especially for younger, adolescent patients. 
      However, human heart valves are limited in availability. Xenogeneic 
      decellularized pig heart valves (dpHV) may serve as alternative. METHODS: The 
      efficacy of DNA reduction processes upon decellularization of heart valves from 
      German Landrace pigs was analyzed by measurements of remaining nucleic acids 
      including proviral porcine endogenous retrovirus (PERV) sequences. Porcine 
      pulmonary heart valves (pPHV) were decellularized by three different protocols 
      and further treated with DNaseI or Benzonase, at varying incubation times. DNA 
      isolated from valve associated muscle (m), valve cusp (c), and pulmonary artery 
      (pa) was monitored by PCR and qRT-PCR using GAPDH and the PERV polymerase (pol) 
      for read-out. RESULTS: Decellularization of pPHV led to a significant reduction 
      of DNA (>99%) which could be further significantly increased for (m) and (pa) by 
      nuclease treatment, reducing proviral PERV pol from approximately 5 x 10(7) to 
      5 x 10(3)  copies/mg in nuclease treated tissues. CONCLUSIONS: Both nucleases 
      demonstrated comparable activities. But DNaseI revealed to be less consistent for 
      PERV, especially at muscular tissue. Noteworthy, remaining proviral sequences are 
      still detectable by PCR; however, due to the absence of the cellular replication 
      machinery the production of infectious particles is not expected. 
      Decellularization and nuclease treatment of pPHV is an efficient procedure to 
      reduce the DNA content including PERV, thus represents a valuable option to 
      increase virus safety independently from the source animal background.
CI  - (c) 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
FAU - Godehardt, Antonia W
AU  - Godehardt AW
AD  - Division of Medical Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.
FAU - Ramm, Robert
AU  - Ramm R
AUID- ORCID: 0000-0002-8556-116X
AD  - Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), 
      Hannover Medical School, Hannover, Germany.
FAU - Gulich, Barbara
AU  - Gulich B
AD  - Division of Medical Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.
FAU - Tonjes, Ralf R
AU  - Tonjes RR
AUID- ORCID: 0000-0001-8727-1935
AD  - Division of Medical Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.
FAU - Hilfiker, Andres
AU  - Hilfiker A
AD  - Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), 
      Hannover Medical School, Hannover, Germany.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20191106
PL  - Denmark
TA  - Xenotransplantation
JT  - Xenotransplantation
JID - 9438793
RN  - 0 (Nucleic Acids)
SB  - IM
MH  - Animals
MH  - Bioprosthesis/adverse effects
MH  - Cell Line
MH  - Endogenous Retroviruses/*pathogenicity
MH  - Heart Valve Prosthesis/*virology
MH  - Heart Valves/*pathology
MH  - Nucleic Acids/*metabolism
MH  - Proviruses/*pathogenicity
MH  - Swine
MH  - Transplantation, Heterologous/adverse effects
OTO - NOTNLM
OT  - PERV
OT  - virus safety
OT  - xenogeneic decellularized heart valve
EDAT- 2019/11/07 06:00
MHDA- 2021/06/09 06:00
CRDT- 2019/11/07 06:00
PHST- 2019/06/19 00:00 [received]
PHST- 2019/10/04 00:00 [revised]
PHST- 2019/10/13 00:00 [accepted]
PHST- 2019/11/07 06:00 [pubmed]
PHST- 2021/06/09 06:00 [medline]
PHST- 2019/11/07 06:00 [entrez]
AID - 10.1111/xen.12565 [doi]
PST - ppublish
SO  - Xenotransplantation. 2020 Mar;27(2):e12565. doi: 10.1111/xen.12565. Epub 2019 Nov 
      6.