Bone Density Assessment Through Sodium Poly-Tungstate Gradient Centrifugation: A Preliminary Study on Decades-Old Human Samples

Bone density is considered one of the many factors influencing bone structure and DNA preservation. For this reason, it is of interest in fields such as anthropology, palaeontology, and genetics. This study describes a method for bone density assessment by gradient centrifugation in Sodium Poly-Tungstate (SPT) solutions (from 2.1 to 2.6 g/cm(3)). Fifty milligrams of bone powder (size range of 20-50 mu m) were used, with an average recovery of 89.9 (IC = 3.3% at 95% of probability). In the first phase of the experiment, the protocol was applied to ten femurs: three exhumed from the WWII mass grave of Ossero, three aged (43-50 years old) femurs from a museum collection and four fresh controls. In the subsequent phase, the analysis was extended to three petrous bones, three metacarpals, and three metatarsals exhumed from the WWII mass grave. The SPT density gradient profiles revealed marked differences among the three femur sample sets: more than 80% of the powder from control femurs was recovered in fractions with a density <= 2.2 g/cm(3), whereas approximately 45% of the femurs from the mass grave showed a density > 2.6 g/cm(3). The remaining three aged femurs displayed peculiar density patterns. Among the other bone types, metatarsals showed the lowest density values, followed by petrous bones and metacarpals. To detect degradation signatures, all nineteen bone powders were also analysed by ATR-FTIR. The femurs from the mass grave exhibited spectral features consistent with mineral recrystallisation and degradation of the organic phase, whereas the other three aged femurs showed peculiar spectral profiles; metacarpals, petrous bones and metatarsals showed intermediate spectra. PCA was applied to SPT and ATR-FTIR data, revealing correlations that support the SPT method as a novel tool for bone quality assessment. Although based on a limited sample size, this preliminary work demonstrates that SPT gradient analysis is an effective, low-cost, rapid and reliable method for assessing bone density, with potential applications in different disciplines studying aged bone samples. Lastly, principal component analysis (PCA) revealed a correlation between bone density and the yield of DNA recovered from the ten femoral specimens.