Background and Aim: Nanobacteria or calcifying nanoparticles (CNP) possesses unique properties such as small size (0.1 to 5.0 microns) and high resistance to heat, most likely are in vivo candidates to initiate and facilitate the pathologic calcification. These spherical organisms are 100 times smaller than ordinary bacteria and are protected by a shell of crystalline carbonate apatite, so they are etiologic factors for extra-skeletal pathological calcification such as kidney and urinary stones. Methods: Thirty kidney stone and twenty urinary stone were collected from patients who had undergone surgery. After powdering the stones and removing the coating of calcite with hydrochloric acid and neutralization with appropriate buffers, culturing these organisms was performed after filtrations the solution through the 2.0 micron filter in DMEM medium. After 40 days, nanobacteria growth was monitored with Scanning Electron Microscopy (SEM). During this period of growth, the turbidity measurement was performed with a spectrophotometer. Results: The nanobacteria growth was observed using spectrophotometer. Results of scanning electron microscope (SEM) showed a spherical shape in size of 33 to 430 nm. Energy Dispersive X-ray spectroscopy (EDS) also showed that these organisms have a calcium coating around itself. Conclusions: The nanobacteria growth was observed using spectrophotometer. Results of scanning electron microscope (SEM) showed a spherical shape in size of 33 to 430 nm. Energy Dispersive X-ray spectroscopy (EDS) also showed that these organisms have a calcium coating around itself.

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