Chemical induced demineralization study in cortical bone

In this work we present a study of demineralization in bovine cortical bone. We selected 9 fresh cortical bone samples from 2 diaphyseal femurs for analysis. Sampleswere demineralized for 24 h, 48 h, 72 h and 96 h using two concentrations of EDTA with different pH: EDTA 0.1M(pH 10, alkaline) and EDTA 0.5M(pH 7.4, neutral). We have employed mu-X-ray fluorescence (mu-XRF) and X- ray diffraction (XRD) to assess the degree of demineralization. EDTA solutions were analyzed for Calcium (Ca) and Phosphorous (P) extractions by Atomic Absorption Spectrophotometry (AAS) and Ion Chromatography (IC), respectively. Results from AAS and IC showed that EDTA 0.5M (pH 7.4) removed two times more Ca and 3 times more P than EDTA 0.1M (pH 10) in the first 24 hours. mu-XRF results presented that EDTA has a high capacity to bind Calcium and Phosphorus. On the other hand, despite the differences in concentration and pH, EDTA did not bind Zn and Sr. Results from XRD showed that EDTA with high concentration had a greater impact to the samples' crystallinity causing a severe damage.