Naturally produced crystals obtained from kidney stones are less injurious to renal tubular epithelial cells than synthetic crystals

Carla Escobar, Karen J. Byer, Saeed R. Khan

Research output: Contribution to journalArticle

Abstract

OBJECTIVE: To determine the differences in cell responses to synthetic and biological crystals of calcium oxalate (CaOx) and brushite MATERIALS AND METHODS: Nephrolithiasis depends on crystal retention within the kidneys, often promoted by crystal attachment to the injured renal epithelium; studies often use various crystals that might be injurious to cells and cause the exposure of crystal binding molecules on cell surfaces, thus promoting crystal attachment and retention. The synthetic crystals used in these studies might be more injurious than the biological crystals naturally produced in the kidneys and that form kidney stones. We exposed the renal epithelial cell line NRK 52E in vitro to CaOx or brushite crystals at 67 or 133 μg/cm2 for 3 or 6 h. Synthetic crystals were purchased and the biocrystals were obtained by pulverizing CaOx and brushite stones. We determined the release of lactate dehydrogenase (LDH), hydrogen peroxide (H2O2) and 8-isoprostane (8-IP), and monocyte chemoattractant protein-1 (MCP-1), as markers of injury, oxidative stress and inflammation, respectively. Cells were also examined after trypan blue staining to determine their membrane integrity. We also examined crystals of CaOx by scanning electron microscopy both in the native state as well as after decalcification. RESULTS: Exposure to both the synthetic and biological crystals resulted in a significant increase in LDH release and trypan blue staining, as a sign of crystal-induced injury. There was increased production of H2O2 and 8-IP, suggesting the development of oxidative stress. In addition MCP-1 production was also significantly increased. However, the synthetic crystals caused significantly higher increases in all the indicators than the biological crystals. CONCLUSIONS: These results indicate that even though both synthetic and naturally produced biocrystals invoke a response from the renal epithelial cells, the latter are significantly less injurious and inflammatory. Exposure to low concentrations of these crystals alone might not invoke an inflammatory response, cause the uncovering of crystal binding molecules on epithelial cell surfaces, and promote crystal attachment and retention.

Original languageEnglish (US)
Pages (from-to)891-897
Number of pages7
JournalBJU International
Volume100
Issue number4
DOIs
StatePublished - Oct 2007
Externally publishedYes

Fingerprint

Kidney Calculi
Calcium Oxalate
8-epi-prostaglandin F2alpha
Epithelial Cells
Kidney
Trypan Blue
Chemokine CCL2
L-Lactate Dehydrogenase
Oxidative Stress
Staining and Labeling
Nephrolithiasis
Wounds and Injuries
Electron Scanning Microscopy
Hydrogen Peroxide
Epithelium
Inflammation
Cell Line
Membranes
dibasic calcium phosphate dihydrate

Keywords

  • Brushite
  • Calcium oxalate
  • Hyperoxaluria
  • Kidney stones
  • Nephrolithiasis
  • Renal injury

ASJC Scopus subject areas

  • Urology

Cite this

Naturally produced crystals obtained from kidney stones are less injurious to renal tubular epithelial cells than synthetic crystals. / Escobar, Carla; Byer, Karen J.; Khan, Saeed R.

In: BJU International, Vol. 100, No. 4, 10.2007, p. 891-897.

Research output: Contribution to journalArticle

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abstract = "OBJECTIVE: To determine the differences in cell responses to synthetic and biological crystals of calcium oxalate (CaOx) and brushite MATERIALS AND METHODS: Nephrolithiasis depends on crystal retention within the kidneys, often promoted by crystal attachment to the injured renal epithelium; studies often use various crystals that might be injurious to cells and cause the exposure of crystal binding molecules on cell surfaces, thus promoting crystal attachment and retention. The synthetic crystals used in these studies might be more injurious than the biological crystals naturally produced in the kidneys and that form kidney stones. We exposed the renal epithelial cell line NRK 52E in vitro to CaOx or brushite crystals at 67 or 133 μg/cm2 for 3 or 6 h. Synthetic crystals were purchased and the biocrystals were obtained by pulverizing CaOx and brushite stones. We determined the release of lactate dehydrogenase (LDH), hydrogen peroxide (H2O2) and 8-isoprostane (8-IP), and monocyte chemoattractant protein-1 (MCP-1), as markers of injury, oxidative stress and inflammation, respectively. Cells were also examined after trypan blue staining to determine their membrane integrity. We also examined crystals of CaOx by scanning electron microscopy both in the native state as well as after decalcification. RESULTS: Exposure to both the synthetic and biological crystals resulted in a significant increase in LDH release and trypan blue staining, as a sign of crystal-induced injury. There was increased production of H2O2 and 8-IP, suggesting the development of oxidative stress. In addition MCP-1 production was also significantly increased. However, the synthetic crystals caused significantly higher increases in all the indicators than the biological crystals. CONCLUSIONS: These results indicate that even though both synthetic and naturally produced biocrystals invoke a response from the renal epithelial cells, the latter are significantly less injurious and inflammatory. Exposure to low concentrations of these crystals alone might not invoke an inflammatory response, cause the uncovering of crystal binding molecules on epithelial cell surfaces, and promote crystal attachment and retention.",
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AU - Escobar, Carla

AU - Byer, Karen J.

AU - Khan, Saeed R.

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N2 - OBJECTIVE: To determine the differences in cell responses to synthetic and biological crystals of calcium oxalate (CaOx) and brushite MATERIALS AND METHODS: Nephrolithiasis depends on crystal retention within the kidneys, often promoted by crystal attachment to the injured renal epithelium; studies often use various crystals that might be injurious to cells and cause the exposure of crystal binding molecules on cell surfaces, thus promoting crystal attachment and retention. The synthetic crystals used in these studies might be more injurious than the biological crystals naturally produced in the kidneys and that form kidney stones. We exposed the renal epithelial cell line NRK 52E in vitro to CaOx or brushite crystals at 67 or 133 μg/cm2 for 3 or 6 h. Synthetic crystals were purchased and the biocrystals were obtained by pulverizing CaOx and brushite stones. We determined the release of lactate dehydrogenase (LDH), hydrogen peroxide (H2O2) and 8-isoprostane (8-IP), and monocyte chemoattractant protein-1 (MCP-1), as markers of injury, oxidative stress and inflammation, respectively. Cells were also examined after trypan blue staining to determine their membrane integrity. We also examined crystals of CaOx by scanning electron microscopy both in the native state as well as after decalcification. RESULTS: Exposure to both the synthetic and biological crystals resulted in a significant increase in LDH release and trypan blue staining, as a sign of crystal-induced injury. There was increased production of H2O2 and 8-IP, suggesting the development of oxidative stress. In addition MCP-1 production was also significantly increased. However, the synthetic crystals caused significantly higher increases in all the indicators than the biological crystals. CONCLUSIONS: These results indicate that even though both synthetic and naturally produced biocrystals invoke a response from the renal epithelial cells, the latter are significantly less injurious and inflammatory. Exposure to low concentrations of these crystals alone might not invoke an inflammatory response, cause the uncovering of crystal binding molecules on epithelial cell surfaces, and promote crystal attachment and retention.

AB - OBJECTIVE: To determine the differences in cell responses to synthetic and biological crystals of calcium oxalate (CaOx) and brushite MATERIALS AND METHODS: Nephrolithiasis depends on crystal retention within the kidneys, often promoted by crystal attachment to the injured renal epithelium; studies often use various crystals that might be injurious to cells and cause the exposure of crystal binding molecules on cell surfaces, thus promoting crystal attachment and retention. The synthetic crystals used in these studies might be more injurious than the biological crystals naturally produced in the kidneys and that form kidney stones. We exposed the renal epithelial cell line NRK 52E in vitro to CaOx or brushite crystals at 67 or 133 μg/cm2 for 3 or 6 h. Synthetic crystals were purchased and the biocrystals were obtained by pulverizing CaOx and brushite stones. We determined the release of lactate dehydrogenase (LDH), hydrogen peroxide (H2O2) and 8-isoprostane (8-IP), and monocyte chemoattractant protein-1 (MCP-1), as markers of injury, oxidative stress and inflammation, respectively. Cells were also examined after trypan blue staining to determine their membrane integrity. We also examined crystals of CaOx by scanning electron microscopy both in the native state as well as after decalcification. RESULTS: Exposure to both the synthetic and biological crystals resulted in a significant increase in LDH release and trypan blue staining, as a sign of crystal-induced injury. There was increased production of H2O2 and 8-IP, suggesting the development of oxidative stress. In addition MCP-1 production was also significantly increased. However, the synthetic crystals caused significantly higher increases in all the indicators than the biological crystals. CONCLUSIONS: These results indicate that even though both synthetic and naturally produced biocrystals invoke a response from the renal epithelial cells, the latter are significantly less injurious and inflammatory. Exposure to low concentrations of these crystals alone might not invoke an inflammatory response, cause the uncovering of crystal binding molecules on epithelial cell surfaces, and promote crystal attachment and retention.

KW - Brushite

KW - Calcium oxalate

KW - Hyperoxaluria

KW - Kidney stones

KW - Nephrolithiasis

KW - Renal injury

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