Molecular modifiers reveal a mechanism of pathological crystal growth inhibition

Crystalline materials are crucial to the function of living organisms, in the shells of molluscs(1-3), the matrix of bone(4), the teeth of sea urchins5, and the exoskeletons of coccoliths(6). However, pathological biomineralization can be an undesirable crystallization process associated with human diseases(7-9). The crystal growth of biogenic, natural and synthetic materials may be regulated by the action of modifiers, most commonly inhibitors, which range from small ions and molecules(10,11) to large macromolecules(12). Inhibitors adsorb on crystal surfaces and impede the addition of solute, thereby reducing the rate of growth(13,14). Complex inhibitor-crystal interactions in biomineralization are often not well elucidated(15). Here we show that two molecular inhibitors of calcium oxalate monohydrate crystallization-citrate and hydroxycitrate-exhibit a mechanism that differs from classical theory in that inhibitor adsorption on crystal surfaces induces dissolution of the crystal under specific conditions rather than a reduced rate of crystal growth. This phenomenon occurs even in supersaturated solutions where inhibitor concentration is three orders of magnitude less than that of the solute. The results of bulk crystallization, in situ atomic force microscopy, and density functional theory studies are qualitatively consistent with a hypothesis that inhibitor-crystal interactions impart localized strain to the crystal lattice and that oxalate and calcium ions are released into solution to alleviate this strain. Calcium oxalate monohydrate is the principal component of human kidney stones(16-19) and citrate is an often-used therapy(20), but hydroxycitrate is not. For hydroxycitrate to function as a kidney stone treatment, it must be excreted in urine. We report that hydroxycitrate ingested by non-stone-forming humans at an often-recommended dose leads to substantial urinary excretion. In vitro assays using human urine reveal that the molecular modifier hydroxycitrate is as effective an inhibitor of nucleation of calcium oxalate monohydrate nucleation as is citrate. Our findings support exploration of the clinical potential of hydroxycitrate as an alternative treatment to citrate for kidney stones.