The Role of Copper in Struvite Formation in the Context of Infection Urinary Stones: A Journey at the Interface of Physics, Chemistry and Microbiology

Infection urinary stones account for approximately 10-15% of all urinary stones worldwide, with a rising incidence observed in recent decades, particularly in countries with a high Socio-Demographic Index (SDI). This trend has been partially attributed to dietary changes, including increased consumption of processed foods. Heavy metals belong to a group of substances, the source of which can be both food and the human environment. Among many heavy metals, in this study, we focus on copper and investigate its influence on the nucleation and growth of struvite crystals, the primary component of infection urinary stones. Experiments were conducted in artificial urine, both in the presence and absence of <i>Proteus mirabilis</i>, a urease-producing bacterium commonly associated with infection urinary stones. In a bacteria-free system, bacterial urease activity was mimicked by the addition of aqueous ammonia solution. Our results demonstrate that the presence of copper in artificial urine induces a slight shift in the struvite crystallization toward lower pH values, indicating that crystal formation initiates earlier compared to a control test. Additionally, the amount of precipitated struvite increases modestly in the presence of copper. Struvite crystals formed in copper-containing artificial urine are larger and exhibit altered habit and morphology. Energy-dispersive X-ray spectroscopy (EDS) and X-ray photoelectron spectroscopy (XPS) analyses confirm that copper does not incorporate into either the bulk or surface structure of the struvite crystals. X-ray diffraction (XRD) data show that struvite remains the sole crystalline phase, consistent with the control samples. Microbiological assays reveal that copper, at the concentrations tested, does not affect the viability of <i>P. mirabilis</i>, indicating an absence of bacteriostatic or bactericidal effects. To elucidate the physicochemical mechanisms underlying copper's influence on nucleation and growth of struvite, speciation analysis of chemical complexes was performed. This revealed the formation of various copper complexes in artificial urine, including Cu(OH)⁺, CuCit^-, CuC₂O₄, Cu(OH)₂, CuHPO₄, Cu(NH₃)²⁺, Cu(NH₃)₂²⁺, and Cu(NH₃)₃²⁺. These chemical complexes modulate the equilibrium and formation of complexes with Mg²⁺ and PO₄^3- (e.g., MgHCit, MgCit^-, MgOH⁺, MgC₂O₄, MgSO₄, MgHPO₄), contributing to the observed shift in struvite crystallization to lower pH values.