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Association of calcium oxalate monohydrate crystals with MDCK cells
C F Verkoelen1, J C Romijn, W C de Bruijn
1Department of Urology, Erasmus University Rotterdam, The Netherlands.
Abstract:
Many factors are presently known which determine the risk of calcium oxalate (CaOx) stone formation in the kidney, although the early events in the pathogenesis of this disease are still to be elucidated. One of these early events is the interaction of intraluminal crystals with the epithelial cells lining the renal tubules. In this study we determined the interaction of approximately 2 microns calcium oxalate monohydrate (COM) crystal with monolayers of Madin-Darby canine kidney (MDCK) cells grown on porous supports in a two-compartment culture system. Crystal-cell interaction studies were performed after the monolayers reached their highest level of gamma-glutamyltranspeptidase (gamma GT) enzyme activity, a marker for brush border development. Technical aspects were evaluated, such as the size and morphology of the crystals and the influence of incubation time, temperature and pH on crystal-cell interaction. Kinetic data demonstrated that an equilibrium between free and associated particles was reached within 30 minutes. Crystal-cell interaction was often associated with cell damage. However, evidence is provided that in an environment that was saturated with calcium oxalate, MDCK cells in an environment that was a certain amount of COM crystals without sustaining measurable injury. After initial attachment to the cell surface, crystals were taken up and subsequently eliminated again from the monolayers. The model system described in this paper provides a tool for detailed studies of processes that are involved in renal cellular handling of luminal COM crystals.
Insights
Researchers studied how calcium oxalate monohydrate (COM) crystals interact with kidney cells. They found cells can handle COM crystals, even taking them up and eliminating them, without significant injury in a saturated environment.
Area of Science:
- Nephrology
- Cell Biology
- Biochemistry
Background:
- Kidney stone formation involves calcium oxalate (CaOx) crystals interacting with renal tubule epithelial cells.
- Early pathogenic events in CaOx stone disease require further elucidation.
- Understanding crystal-cell interactions is crucial for kidney stone research.
Purpose of the Study:
- To investigate the interaction between calcium oxalate monohydrate (COM) crystals and Madin-Darby canine kidney (MDCK) cell monolayers.
- To evaluate the technical aspects influencing crystal-cell interactions, including crystal properties and environmental factors.
- To establish a model system for studying renal cellular handling of COM crystals.
Main Methods:
- Utilized a two-compartment culture system with MDCK cell monolayers on porous supports.
- Assessed crystal-cell interactions after brush border development, indicated by gamma-glutamyltranspeptidase (gamma GT) activity.
- Examined the influence of incubation time, temperature, and pH on crystal-cell interactions.
- Evaluated crystal size, morphology, and uptake/elimination dynamics.
Main Results:
- An equilibrium between free and associated COM crystals was reached within 30 minutes.
- Crystal-cell interaction could lead to cell damage, but MDCK cells showed resilience in a calcium oxalate-saturated environment.
- MDCK cells demonstrated the ability to internalize and subsequently eliminate COM crystals.
- The study identified key technical parameters affecting crystal-cell interactions.
Conclusions:
- The developed model system allows for detailed investigation of renal cellular handling of luminal COM crystals.
- MDCK cells can manage COM crystal exposure, suggesting potential mechanisms for renal stone prevention.
- This research provides a valuable tool for understanding the initial stages of calcium oxalate nephrolithiasis pathogenesis.