Contribution of organic material to the ion composition of bone

Studies of bone mineral ranging from cadaveric analysis to the use of high-resolution ion microprobe with secondary ion mass spectroscopy (SIMS) have concluded that bone is rich in sodium and potassium relative to calcium. Exposure of bone to acid conditions either in vitro or in vivo leads to an exchange of hydrogen ions for sodium and potassium buffering the acidity of the medium or blood, respectively. Whether these monovalent ions reside within the mineral or organic phases of bone has never been determined. To determine the contribution of organic material to bone ion composition, we dissected calvariae from 4- to 6-day-old mice, removed organic material of some with hydrazine (Hydr), and prepared all bones for analysis using a high-resolution scanning ion microprobe coupled to a secondary ion mass spectrometer. We found that in non-Hydr-treated calvariae (Ctl) there was far more surface sodium and potassium than calcium (Na-23/Ca-40 = 15.7 + 1.9, ratio of counts of detected secondary ions, mean + 95% CI, K-39/Ca-40 = 44.0 + 1.5). Removal of organic material with hydrazine (Hydr) led to a marked fall in the ratio of sodium to calcium and potassium to calcium (Na-23/Ca-40 = 5.9 + 1.4, p < 0.025 vs. respective Ctl and K-39/Ca-40 = 1.1 + 1.5,p < 0.001 vs. respective Ctl). Similarly, when examining the cross-section of the calvariae there was more sodium and potassium than calcium (Na-23/Ca-40 = 8.6 + 1.6, K-39/Ca-40 = 26.7 + 1.8). Treatment with Hydr again caused a marked fall in both ratios (Na-23/Ca-40 = 0.3 + 1.6, p < 0.001 vs, respective Ctl and K-39/Ca-40 = 0.02 + 1.9, p < 0.001 vs, respective Ctl). Thus,,within bone the organic material contains the majority of the sodium and potassium. This suggests that the organic material in bone and not the mineral itself is responsible for the acute buffering of the additional hydrogen ions during metabolic acidosis.