Calcium oxalate crystals in fetal bovine serum: Implications for cell culture, phagocytosis and biomineralization studies in vitro

Pedraza, Claudio E.; Chien, Y.-C.; McKee, Marc D.

doi:10.1002/jcb.21515

Cited by 21 publications

(17 citation statements)

References 46 publications

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“…19,20 In fact, studies on COM microcrystals also showed similar results. 19,21 However, in this study, the blank values of cell viability were those after 24 hours of incubation without microcrystal addition. The experiments for different adhesion times (24,18,12,6, and 2 hours) started at 0, 6, 12, 18, and 22 hours, respectively.…”

Section: Morphology Change In Vero Cells Before and After Injurymentioning

confidence: 69%

Interaction between submicron COD crystals and renal epithelial cells

Peng¹,

Ouyang²,

Yao³

et al. 2012

IJN

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Objectives: This study aims to investigate the adhesion characteristics between submicron calcium oxalate dihydrate (COD) with a size of 150 ± 50 nm and African green monkey kidney epithelial cells (Vero cells) before and after damage, and to discuss the mechanism of kidney stone formation. Methods: Vero cells were oxidatively injured by hydrogen peroxide to establish a model of injured cells. Scanning electron microscopy was used to observe Vero-COD adhesion. Inductively coupled plasma emission spectrometry was used to quantitatively measure the amount of adhered COD microcrystals. Nanoparticle size analyzer and laser scanning confocal microscopy were performed to measure the change in the zeta potential on the Vero cell surface and the change in osteopontin expression during the adhesion process, respectively. The level of cell injury was evaluated by measuring the changes in malonaldehyde content, and cell viability during the adhesion process. Results: The adhesion capacity of Vero cells in the injury group to COD microcrystals was obviously stronger than that of Vero cells in the control group. After adhesion to COD, cell viability dropped, both malonaldehyde content and cell surface zeta potential increased, and the fluorescence intensity of osteopontin decreased because the osteopontin molecules were successfully covered by COD. Submicron COD further damaged the cells during the adhesion process, especially for Vero cells in the control group, leading to an elevated amount of attached microcrystals. Conclusion: Submicron COD can further damage injured Vero cells during the adhesion process. The amount of attached microcrystals is proportional to the degree of cell damage. The increased amount of microcrystals that adhered to the injured epithelial cells plays an important role in the formation of early-stage kidney stones.

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Section: Morphology Change In Vero Cells Before and After Injurymentioning

confidence: 69%

Interaction between submicron COD crystals and renal epithelial cells

Peng¹,

Ouyang²,

Yao³

et al. 2012

IJN

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“…Recent in vitr o studies showed that complete transformation is possible in time spans of the order of 5–75 hours [39]–[41]. This process may easily arrive at complete replacement of the dihydrate by the monohydrate phase in stones in vivo and depends on the residence time in the urinary tract.…”

Section: Discussionmentioning

confidence: 99%

Kidney Stones in Primary Hyperoxaluria: New Lessons Learnt

et al. 2013

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To investigate potential differences in stone composition with regard to the type of Primary Hyperoxaluria (PH), and in relation to the patient’s medical therapy (treatment naïve patients versus those on preventive medication) we examined twelve kidney stones from ten PH I and six stones from four PH III patients. Unfortunately, no PH II stones were available for analysis. The study on this set of stones indicates a more diverse composition of PH stones than previously reported and a potential dynamic response of morphology and composition of calculi to treatment with crystallization inhibitors (citrate, magnesium) in PH I. Stones formed by PH I patients under treatment are more compact and consist predominantly of calcium-oxalate monohydrate (COM, whewellite), while calcium-oxalate dihydrate (COD, weddellite) is only rarely present. In contrast, the single stone available from a treatment naïve PH I patient as well as stones from PH III patients prior to and under treatment with alkali citrate contained a wide size range of aggregated COD crystals. No significant effects of the treatment were noted in PH III stones. In disagreement with findings from previous studies, stones from patients with primary hyperoxaluria did not exclusively consist of COM. Progressive replacement of COD by small COM crystals could be caused by prolonged stone growth and residence times in the urinary tract, eventually resulting in complete replacement of calcium-oxalate dihydrate by the monohydrate form. The noted difference to the naïve PH I stone may reflect a reduced growth rate in response to treatment. This pilot study highlights the importance of detailed stone diagnostics and could be of therapeutic relevance in calcium-oxalates urolithiasis, provided that the effects of treatment can be reproduced in subsequent larger studies.

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“…Different growth conditions frequently used for the study of calcium oxalate commonly show at least some crystals of both the monohydrate and dihydrate forms. In an example of an equilibrated state, calcium oxalate crystals found in the major sources of commercially available fetal bovine serum routinely supplied as a growth-and differentiation-inducing supplement for cells in culture have been reported to sequester numerous proteins from the serum, and most of the COD crystals adopted the {101} tetragonal di-pyramidal shape (22). In this same study, Western blotting for OPN from protein extracts derived from the crystals separated from the fetal bovine serum confirmed likewise in this case the presence of abundant OPN bound by the crystals.…”

Section: Discussionmentioning

confidence: 99%

“…Substantial elegant work has been performed on COM growth and the effect of citrate and peptides/proteins as crystal growth modifiers. Some macromolecules, including urinary osteopontin (OPN), contain polyanionic regions and net negative charges that have been shown to inhibit calcium oxalate crystallization (14, 18 -21) and influence calcium oxalate growth in favor of COD (3,10,22,23). Although there appears to be preferential inhibition of COM, several studies present evidence for higher affinity of OPN binding to COD, and here we investigate this further to show the effects of OPN (and a peptide of OPN) on COD crystal growth and provide information on peptide/protein occlusion that might facilitate crystal dissolution as originally proposed by Ryall et al (see below) (23,24).…”

mentioning

confidence: 99%