Abstract
Our previous report showed that uropathogenic bacteria, e.g., Escherichia coli, are commonly found inside the nidus of calcium oxalate (CaOx) kidney stones and may play pivotal roles in stone genesis. The present study aimed to prove this new hypothesis by direct examining CaOx lithogenic activities of both Gram-negative and Gram-positive bacteria. CaOx was crystallized in the absence (blank control) or presence of 105 CFU/ml E. coli, Klebsiella pneumoniae, Staphylococcus aureus, or Streptococcus pneumoniae. Fragmented red blood cell membranes and intact red blood cells were used as positive and negative controls, respectively. The crystal area and the number of aggregates were measured to initially screen for effects of bacteria on CaOx crystal growth and aggregation. The data revealed that all the bacteria tested dramatically increased the crystal area and number of crystal aggregates. Validation assays (spectrophotometric oxalate-depletion assay and an aggregation–sedimentation study) confirmed their promoting effects on both growth (20.17 ± 3.42, 17.55 ± 2.27, 16.37 ± 1.38, and 21.87 ± 0.85 % increase, respectively) and aggregation (57.45 ± 2.08, 51.06 ± 5.51, 55.32 ± 2.08, and 46.81 ± 3.61 % increase, respectively) of CaOx crystals. Also, these bacteria significantly enlarged CaOx aggregates, with the diameter greater than the luminal size of distal tubules, implying that tubular occlusion might occur. Moreover, these bacterial effects were dose-dependent and specific to intact viable bacteria, not intact dead or fragmented bacteria. In summary, intact viable E. coli, K. pneumoniae, S. aureus, and S. pneumoniae had significant promoting effects on CaOx crystal growth and aggregation. This functional evidence supported the hypothesis that various types of bacteria can induce or aggravate metabolic stone disease, particularly the CaOx type.
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Acknowledgments
We are grateful to Somporn Srifuengfung and Chanwit Tribuddharat for their advice on microbiological techniques, and to Sakdithep Chaiyarit and Teerada Homvises for their technical support. This study was supported by the Office of the Higher Education Commission and Mahidol University under the National Research Universities Initiative (to V.T.), the Thailand Research Fund (MRG5380018 to S.C. and RTA5380005 to V.T.), and the Faculty of Medicine Siriraj Hospital. W.C. is supported by the Royal Golden Jubilee PhD Program, and V.T. is also supported by the Chalermphrakiat grant and the Faculty of Medicine Siriraj Hospital.
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Chutipongtanate, S., Sutthimethakorn, S., Chiangjong, W. et al. Bacteria can promote calcium oxalate crystal growth and aggregation. J Biol Inorg Chem 18, 299–308 (2013). https://doi.org/10.1007/s00775-012-0974-0
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DOI: https://doi.org/10.1007/s00775-012-0974-0