Autologous bone used for the reconstruction of bone defects remains still the gold standard, but is accompanied by donor site morbidities. Common devitalisation methods to process allografts often lead to diminished biomechanical properties. Regarding previous studies, High Hydrostatic Pressure (HHP) treatment may represent a gentle alternative to already existing methods. The effect of HHP on trabecular bone blocks was analysed by quantification of DNA-fragmentation, gene expression, protein integrity and histological stainings. The collected data demonstrate that HHP treatments with 250 and 300 MPa show devitalizing effects on human trabecular bone and therefore may be a favorable alternative to the existing physical-chemical devitalizing processing methods for allografts. In addition, treatment with HHP showed that despite devitalization the matrix integrity could be well preserved. However, further detailed in vivo studies aimed at investigating host responses as well as the revitalization of allograft materials are essential to implement HHD-based treatments into the clinical practice.