Aim: to analyze three-dimensional micro- and macro-structural changes in normal and osteoporotic human bone processed into allografts for transplantation.

Method: ssNMR, SEM, μCT-MIMICS and TGA were used for delineating the ultra-structural property of cortical and cancellous bone.

Findings: Altered balance in water content and association, collagen tertiary structure, and crystalline order in normal and osteoporotic bone (ssNMR).

Pasteurization, alcohol treatment and radiation affect bone through bound water integrity resulting in shifted HU (μCT-MIMICS®), which could make bone weaker.

Cross-correlation between probes used quantitatively demonstrated that protocol adopted for bone processing could play a crucial role in allograft preparation.