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Lyophilization Does Not Inactivate Infectious Retrovirus in Systemically Infected Bone and Tendon Allografts

Matthew J. Crawford, DO, PhD^*, Cheryl L. Swenson, DVM, PhD, Steven P. Arnoczky, DVM^*,,, Jeremy O’Shea^* and Herbert Ross, DO^*

From the ^* College of Osteopathic Medicine, the Department of Pathobiology and Diagnostic Investigation, and the Laboratory for Comparative Orthopaedic Research, Michigan State University, East Lansing, Michigan

Address correspondence to Steven P. Arnoczky, DVM, Director, Laboratory for Comparative Orthopaedic Research, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824 (e-mail: arnoczky{at}cvm.msu.edu).

Background: A review of multiple transplantations of human immunodeficiency virus–infected musculoskeletal allografts found that recipients of lyophilized (freeze-dried) bone or tendon from an infected donor all tested negative for human immunodeficiency virus. The finding that 75% of the recipients of fresh-frozen bone from the same donor contracted human immunodeficiency virus has led to speculation that freeze-drying may render retroviral-infected musculoskeletal allografts noninfectious.

Hypothesis: Lyophilization does not inactivate retrovirus in systemically infected bone and tendon.

Study Design: Controlled laboratory study.

Methods: Tendons and cortical bone segments from cats systemically infected with feline leukemia virus were used in this study. Feline embryonic fibroblast cells were cultured in the presence of fresh-frozen or freeze-dried cortical bone or tendon segments. At each passage, feline leukemia virus p27 antigen was measured in media by enzyme-linked immunosorbent assay, and feline leukemia virus (pro)viral nucleic acids were quantified by real-time quantitative polymerase chain reaction in the DNA extracted from cells.

Results: Enzyme-linked immunosorbent assay results and quantitative polymerase chain reaction results demonstrated retroviral antigen and proviral DNA in all cultured cell replicates after exposure to fresh-frozen or freeze-dried bones or tendons.

Conclusion: Freeze-drying (lyophilization) of retroviral-infected cortical bone and tendon does not inactivate retrovirus.

Clinical Relevance: These results conclusively demonstrate that freeze-drying should not be relied on to inactivate infectious retrovirus in systemically infected musculoskeletal allografts.

Key Words: allograft • lyophilization • retrovirus • tendon • bone

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