Physiochemical Dissolution Governs Early Modifications in Acid-Exposed Murine Bone with Long-term Recovery

Abstract

Metabolic acidosis (MA), a disease affecting millions annually, is clinically characterized by a decrease in systemic pH and bicarbonate. MA has been shown to have skeletal consequences, such as increased bone loss and fractures. However, the mechanisms impacting the mechanics of bone remain unknown. To determine this, we characterized the skeletal phenotypes resulting from two independent regimes of ammonium chloride (NH4Cl) acid-dosing in skeletally mature CD-1 mice for 1, 3, 7, and 14 days to establish the temporal progression of materials-level skeletal defects. The classical, flat-dose model resulted in a temporary decrease in blood pH but did not reduce blood bicarbonate levels. Minimal alterations were observed in bone composition, structure, mechanics, and cellular behavior. However, the graded administration of NH4Cl maintained MA for up to 14 days. This resulted in decreased bone mineral content, collagen organization, and bone volume while increasing mineral crystallinity at early time points followed by a return to baseline. Changes to the matrix lead to a potential decrease in bone toughness at early time points, which may explain the increased fracture risk seen clinically. However, there were no changes in bone formation rate nor osteoclast number or activity during any time point. After 14 days, the bones were fully recovered even though the mice were still under acidotic conditions. Ultimately, we conclude that physicochemical dissolution is the main mechanism for early changes in murine bone, which is influenced by the dosing of exogenous acid loading. This knowledge will aid the discovery of treatments that prevent acidosis-induced bone dissolution.