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Wellcome open research2017; 2; 60; doi: 10.12688/wellcomeopenres.12176.2

Identification of Equid herpesvirus 2 in tissue-engineered equine tendon.

Abstract: Incidental findings of virus-like particles were identified following electron microscopy of tissue-engineered tendon constructs (TETC) derived from equine tenocytes. We set out to determine the nature of these particles, as there are few studies which identify virus in tendons , and their presence could have implications for tissue-engineering using allogenic grafts. Virus particles were identified in electron microscopy of TETCs. Virion morphology was used to initially hypothesise the virus identity.  Next generation sequencing was implemented to identify the virus. A pan herpesvirus PCR was used to validate the RNASeq findings using an independent platform. Histological analysis and biochemical analysis was undertaken on the TETCs. Morphological features suggested the virus to be either a retrovirus or herpesvirus. Subsequent next generation sequencing mapped reads to Equid herpesvirus 2 (EHV2). Histological examination and biochemical testing for collagen content revealed no significant differences between virally affected TETCs and non-affected TETCs. An independent set of equine superficial digital flexor tendon tissue (n=10) examined using designed primers for specific EHV2 contigs identified at sequencing were negative. These data suggest that EHV is resident in some equine tendon. EHV2 was demonstrated in equine tenocytes for the first time; likely from infection. The presence of EHV2 could have implications to both tissue-engineering and tendinopathy.
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Publication Date: 2017-10-17 PubMed ID: 29152595PubMed Central: PMC5664983DOI: 10.12688/wellcomeopenres.12176.2Google Scholar: Lookup
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Summary

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This research paper delves into the unexpected identification of Equid herpesvirus 2 (EHV2) in tissue-engineered equine tendon constructs (TETC) created from equine tenocytes. Despite not showing any significant biochemical or histological differences with non-infected tissues, the presence of the virus might have potential consequences for tissue engineering and treatment of tendinopathy.

Methodology

  • The discovery of virus-like particles in the tissue-engineered tendon constructs began as an incidental discovery under an electron microscope.
  • The team employed virion morphology to come up with an initial assumption about the identity of the virus.
  • The next generation sequencing was used to definitively identify the virus particles. An independent platform was also used to confirm the RNASeq results. This was executed using a pan herpesvirus PCR.
  • Equivalent histological analysis and biochemical examinations were conducted on the TETCs to compare and understand any differences between non-infected and virally infected TETCs.
  • Additional checks involved investigating equine superficial digital flexor tendon tissue from by utilising primers designed for specific EHV2 contigs.

Findings

  • The viral particles were initially assumed to be either a herpesvirus or retrovirus based on their morphology.
  • Following the engagement of next generation sequencing, the virus was authenticated as EHV2.
  • Upon comparing the virally infected TETCs with non-infected ones, no significant differences were observed in terms of collagen content or histological attributes.
  • All ten of the other samples of equine superficial digital flexor tendon tissue investigated were found to be negative for the virus.
  • The data suggests that EHV2 resides in some equine tendon tissue, marking the first time it has been found in tenocytes.

Implications

  • The findings introduce the possibility that EHV2 might have implications for tissue engineering involving equine tendons and the treatment of tendinopathy conditions.
  • While further study is required, the identification of EHV2 in equine tissues might affect choices in the use of autologous or allogenous grafts in tissue engineering.

Cite This Article

APA
Wardle R, Pullman JA, Haldenby S, Ressel L, Pope M, Clegg PD, Radford A, Stewart JP, Al-Saadi M, Dyer P, Peffers MJ. (2017). Identification of Equid herpesvirus 2 in tissue-engineered equine tendon. Wellcome Open Res, 2, 60. https://doi.org/10.12688/wellcomeopenres.12176.2

Publication

Wellcome open research
ISSN: 2398-502X
NlmUniqueID: 101696457
Country: England
Language: English
Volume: 2
Pages: 60
PII: 60

Researcher Affiliations

Wardle, Roisin
  • Institute of Veterinary Science, University of Liverpool, Leahurst Campus, Chester High Road, Neston, Wirral, UK.
Pullman, Jane A
  • Centre for Genomic Research, Institute of Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool, UK.
Haldenby, Sam
  • Centre for Genomic Research, Institute of Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool, UK.
Ressel, Lorenzo
  • Institute of Veterinary Science, University of Liverpool, Leahurst Campus, Chester High Road, Neston, Wirral, UK.
Pope, Marion
  • Institute of Veterinary Science, University of Liverpool, Leahurst Campus, Chester High Road, Neston, Wirral, UK.
Clegg, Peter D
  • Institute of Ageing and Chronic Disease, University of Liverpool, William Henry Duncan Building, West Derby Street, Liverpool, UK.
Radford, Alan
  • Institute of Infection and Global Health, Department of Infection Biology, University of Liverpool, West Derby Street, Liverpool , UK.
Stewart, James P
  • Institute of Infection and Global Health, Department of Infection Biology, University of Liverpool, West Derby Street, Liverpool , UK.
Al-Saadi, Mohammed
  • Institute of Infection and Global Health, Department of Infection Biology, University of Liverpool, West Derby Street, Liverpool , UK.
Dyer, Philip
  • Institute of Veterinary Science, University of Liverpool, Leahurst Campus, Chester High Road, Neston, Wirral, UK.
Peffers, Mandy J
  • Institute of Ageing and Chronic Disease, University of Liverpool, William Henry Duncan Building, West Derby Street, Liverpool, UK.

Grant Funding

  • Wellcome Trust
  • G0900753 / Medical Research Council
  • MR/K002279/1 / Medical Research Council
  • MR/P020941/1 / Medical Research Council

Conflict of Interest Statement

Competing interests: No competing interests were disclosed.

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