Validation of radiation sterilization dose for lyophilized amnion and bone grafts.

A limited number of grafts produced in one batch is the main constrain to validate radiation sterilization dose of amnion and bone grafts according to ISO standard. The validation experiments done were according to ISO 13409 with a slight modification in sampling method. The experiments were carried out three times by using 20 samples each, 10 for bio-burden enumeration and 10 for sterility test at verification dose. The average bio-burden with sample item portion (SIP) = 1 for amnion membranes were 98, 50 and 69 cfu respectively and 0 cfu for bone grafts. Verification dose experiments, were done at doses of 2.90 kGy for bone grafts and 5.13 kGy for amnion grafts and the results of sterility tests showed that amnion grafts got one positive and bone grafts got 0 positive. The results met the requirements of ISO 13409 so that the radiation sterilization dose, at sterility assurance level of 10(-6) was 25 kGy for both amnion and bone grafts. Viral contamination was excluded in this experiment.