A rapid method for the generation of uniform acellular bone explants: a technical note

Background: Bone graft studies lack standardized controls. We aim
to present a quick and reliable method for the intra-operative
generation of acellular bone explants. Methods: Therefore, ovine
cancellous bone explants from the iliac crest were prepared and
used to test several methods for the induction of cell death. Over
night heat inactivation was used as positive treatment control,
methods to be investigated included UV light, or X-ray exposure,
incubation in a hypotonic solution (salt-free water) and a short
cycle of repeated freezing and thawing. Results: Viability of
treated and 2 days cultured bone explants was investigated by
lactate dehydrogenase assay. Non-treated cultured control explants
maintained around 50% osteocyte viability, while osteocyte survival
after the positive treatment control was abolished. The most
dramatic loss in cell viability, together with a low standard
deviation, was a repeated cycle of freezing and thawing.
Conclusions: To summarize, we present a freeze-thaw method for the
creation of acellular bone explants, which is easy to perform, not
time-consuming and provides consistent results.