An in vitro ultrastructural study of infectious kidney stone genesis

Abstract

A ureolytic strain of Proteus mirabilis, isolated from a patient with infectious kidney stones, produced struvite (MgNH4PO4 X 6 H2O) and apatite [Ca10(PO4)6CO3] crystals in vitro when grown in artificial urine. Surface-attached crystals were encased in a slime-like layer. Scanning electron microscopy revealed that surfaces submerged in the artificial urine were colonized by P. mirabilis. Bacteria-associated crystals appeared soon after colonization and eventually became coated with an amorphous substance. Energy-dispersive X-ray analysis of these crystals revealed the presence of Mg, Ca, and P which are major components of struvite and apatite. Transmission electron microscopy of surface scrapings revealed that the glycocalyx of P. mirabilis contained a large number of crystals. Based on these observations and previous work, a theory for infectious renal calculogenesis is proposed. The kidney is initially colonized by invading ureolytic pathogens. These pathogens secrete copious amounts of glycocalyx which facilitates adhesion of the organisms to the kidney, provides protection for these bacteria, and serves to bind struvite and apatite crystals that result from bacterial urease activity. Growth of these calcified microcolonies into mature stones is characterized by continued bacterial growth, incorporation of urinary mucoproteins into the matrix along with bacterial glycocalyx, and a continued deposition of struvite and apatite crystals due to the high pH. The mature stone, in effect, represents an enlarged "fossilized" bacterial microcolony.