Comparison of the efficacy of virus inactivation methods in allogeneic avital bone tissue transplants

Several procedures for inactivating viruses are used presently in the context of bone tissue transplants. Common methods used are gamma irradiation (25 kGy), treatment with moist heat (82.5 degrees C/15 min., lobator-sd2-system) as well as chemical sterilisation using peracetic acid-ethanol treatment (PES, 2% peracetic acid, 96% ethanol, Aqua [2:1:1], 200 mbar, agitation, 4 hours). Based on national and international guidelines, we tested the antivirucidal effectiveness of these methods in human bone transplants. Three enveloped viruses: human immunodeficiency virus type 2 (HIV-2), pseudorabies virus (PRV), bovine virus diarrhoea virus (BVDV), and three non-enveloped viruses were used: hepatitis A virus (HAV), poliovirus (PV-1), porcine/bovine parvovirus (PPV, BPV). Defatted spongiosa cuboids served as model in chemical treatment experiments, while cortical diaphyses were used in gamma irradiation experiments, and the effects of thermal treatment were tested in prepared femoral heads. The log(10) reduction was measured by cytopathogenic effects after virus titration (TCID(50)/mL). A dose of at least 33.9 kGy (bone model) at -30 +/- 5 degrees C was necessary to achieve a sufficient reduction (4 log(10) steps) of BPV, the most resistant one of all viruses investigated. Thermal treatment as well as PES treatment led to a reduction of virus titres by more than 4 log(10). Only HAV showed a reduction below 4 log(10) (2.87) with PES. After validation of the defatting step included for HAV-infected cells, a HAV-reduction of over 7 log(10) was found. All three sterilisation methods tested are recommended for bone transplant sterilisation, but only provided that additional safety measures (anamnestic informations, infectious serology, PCR in case of multiorgan donors) are taken.