High-Throughput in vitro Gel-Based Plate Assay to Screen for Calcium Oxalate Stone Inhibitors

2021 ◽  
pp. 1-7
Author(s):  
John A. Chmiel ◽  
Gerrit A. Stuivenberg ◽  
Abdulaziz Alathel ◽  
Jaswanth Gorla ◽  
Bernd Grohe ◽  
...  
Keyword(s):  
Calcium Oxalate ◽  
High Throughput ◽  
Current Knowledge ◽  
Medical Condition ◽  
Urinary Calculi ◽  
Sodium Oxalate ◽  
Crystal Formation ◽  
Test Substance ◽  
Caox Crystal

<b><i>Objective:</i></b> Kidney stones are a common medical condition that is increasing in prevalence worldwide. Approximately, ∼80% of urinary calculi are composed of calcium oxalate (CaOx). There is a growing interest toward identifying therapeutic compounds that can inhibit the formation of CaOx crystals. However, some chemicals (e.g., antibiotics and bacterial metabolites) may directly promote crystallization. Current knowledge is limited regarding crystal promoters and inhibitors. Thus, we have developed an in vitro gel-based diffusion model to screen for substances that directly influence CaOx crystal formation. <b><i>Materials and Methods:</i></b> We used double diffusion of sodium oxalate and calcium chloride-loaded paper disks along an agar medium to facilitate the controlled formation of monohydrate and dihydrate CaOx crystals. A third disk was used for the perpendicular diffusion of a test substance to assess its influence on CaOx crystal formation. <b><i>Results:</i></b> We confirmed that citrates and magnesium are effective inhibitors of CaOx crystals. We also demonstrated that 2 strains of uropathogenic <i>Escherichia coli</i> are able to promote crystal formation. While the other tested uropathogens and most antibiotics did not change crystal formation, ampicillin was able to reduce crystallization. <b><i>Conclusion:</i></b> We have developed an inexpensive and high-throughput model to evaluate substances that influence CaOx crystallization.

scholarly journals Small intestine resection increases oxalate and citrate transporter expression and calcium oxalate crystal formation in rat hyperoxaluric kidneys

2020 ◽  
Vol 134 (19) ◽  
pp. 2565-2580
Author(s):  
Yi-Shiou Tseng ◽  
Wen-Bin Wu ◽  
Yun Chen ◽  
Feili Lo Yang ◽  
Ming-Chieh Ma
Keyword(s):  
Small Intestine ◽  
Calcium Oxalate ◽  
Cell Injury ◽  
Crystal Formation ◽  
Crystal Deposition ◽  
Tubular Cells ◽  
Citrate Transporter ◽  
Caox Crystal

Abstract Short bowel (SB) increases the risk of kidney stones. However, the underlying mechanism is unclear. Here, we examined how SB affected renal oxalate and citrate handlings for in vivo hyperoxaluric rats and in vitro tubular cells. SB was induced by small intestine resection in male Wistar rats. Sham-operated controls had no resection. After 7 days of recovery, the rats were divided into control, SB (both fed with distilled water), ethylene glycol (EG), and SB+EG (both fed with 0.75% EG for hyperoxaluric induction) groups for 28 days. We collected the plasma, 24 h of urine, kidney, and intestine tissues for analysis. Hypocitraturia was found and persisted up to 28 days for the SB group. Hypocalcemia and high plasma parathyroid hormone (PTH) levels were found in the 28-day SB rats. SB aggravated EG-mediated oxalate nephropathy by fostering hyperoxaluria and hypocitraturia, and increasing the degree of supersaturation and calcium oxalate (CaOx) crystal deposition. These effects were associated with renal up-regulations of the oxalate transporter solute carrier family 26 (Slc26)a6 and citrate transporter sodium-dependent dicarboxylate cotransporter-1 (NaDC-1) but not Slc26a2. The effects of PTH on the SB kidneys were then examined in NRK-52E tubular cells. Recombinant PTH attenuated oxalate-mediated cell injury and up-regulated NaDC-1 via protein kinase A (PKA) activation. PTH, however, showed no additive effects on oxalate-induced Slc26a6 and NaDC-1 up-regulation. Together, these results demonstrated that renal NaDC-1 upregulation-induced hypocitraturia weakened the defense against Slc26a6-mediated hyperoxaluria in SB kidneys for excess CaOx crystal formation. Increased tubular NaDC-1 expression caused by SB relied on PTH.

scholarly journals The Protective Roles of Estrogen Receptor β in Renal Calcium Oxalate Crystal Formation via Reducing the Liver Oxalate Biosynthesis and Renal Oxidative Stress-Mediated Cell Injury

2019 ◽  
Vol 2019 ◽  
pp. 1-17 ◽  
Author(s):  
Wei Zhu ◽  
Zhijian Zhao ◽  
Fu-Ju Chou ◽  
Li Zuo ◽  
Tongzu Liu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Estrogen Receptor ◽  
Nadph Oxidase ◽  
Calcium Oxalate ◽  
Cell Lines ◽  
Ros Production ◽  
Crystal Deposition ◽  
Caox Crystal

Females develop kidney stones less frequently than males do. However, it is unclear if this gender difference is related to altered estrogen/estrogen receptor (ER) signaling. Here, we found that ER beta (ERβ) signals could suppress hepatic oxalate biosynthesis via transcriptional upregulation of the glyoxylate aminotransferase (AGT1) expression. Results from multiple in vitro renal cell lines also found that ERβ could function via suppressing the oxalate-induced injury through increasing the reactive oxygen species (ROS) production that led to a decrease of the renal calcium oxalate (CaOx) crystal deposition. Mechanism study results showed that ERβ suppressed oxalate-induced oxidative stress via transcriptional suppression of the NADPH oxidase subunit 2 (NOX2) through direct binding to the estrogen response elements (EREs) on the NOX2 5′ promoter. We further applied two in vivo mouse models with glyoxylate-induced renal CaOx crystal deposition and one rat model with 5% hydroxyl-L-proline-induced renal CaOx crystal deposition. Our data demonstrated that mice lacking ERβ (ERβKO) as well as mice or rats treated with ERβ antagonist PHTPP had increased renal CaOx crystal deposition with increased urinary oxalate excretion and renal ROS production. Importantly, targeting ERβ-regulated NOX2 with the NADPH oxidase inhibitor, apocynin, can suppress the renal CaOx crystal deposition in the in vivo mouse model. Together, results from multiple in vitro cell lines and in vivo mouse/rat models all demonstrate that ERβ may protect against renal CaOx crystal deposition via inhibiting the hepatic oxalate biosynthesis and oxidative stress-induced renal injury.

The Effect of Ethane-1-Hydroxy-1,1-Diphosphonate (EHDP) on Calcium Oxalate Crystalluria in Recurrent Renal Stone-Formers

1974 ◽  
Vol 47 (1) ◽  
pp. 13-22 ◽  
Author(s):  
W. G. Robertson ◽  
M. Peacock ◽  
R. W. Marshall ◽  
F. Knowles
Keyword(s):  
Calcium Oxalate ◽  
Stone Formation ◽  
Basal Diet ◽  
Sodium Oxalate ◽  
Calcium Oxalate Crystals ◽  
Calcium Oxalate Dihydrate ◽  
Calcium Oxalate Stone ◽  
Oxalate Crystals ◽  
Stone Formers

1. The volume, size and type of calcium oxalate crystals excreted in the urine of a group of patients with recurrent ‘idiopathic’ stones were studied on a controlled basal diet, after an oral supplement of sodium oxalate and after oral administration of ethane-1-hydroxy-1,1-diphosphonate (EHDP) for 4 weeks. 2. Before administration of EHDP the stone-formers passed the large crystals and aggregates of calcium oxalate dihydrate characteristic of recurrent calcium oxalate stone-formers. For the same level of urine saturation and crystalluria EHDP caused a significant reduction in the proportion of large crystals and aggregates excreted. Studies by light-microscopy confirmed that EHDP caused a striking change in the size and habit of calcium oxalate crystals in some but not all of the urine samples examined. 3. The decrease in average crystal size during the administration of EHDP was attributed to the observed increase in the ability of urine to inhibit the growth and aggregation of calcium oxalate crystals as measured by a growth system in vitro. 4. The possible use of EHDP as a therapeutic agent in the treatment of calcium oxalate stone-formation is discussed.

scholarly journals The Role of Extracellular Matrix Components in Dentin Mineralization

1991 ◽  
Vol 2 (3) ◽  
pp. 369-387 ◽  
Author(s):  
Adele L. Boskey
Keyword(s):  
Extracellular Matrix ◽  
Current Knowledge ◽  
Bone Matrix ◽  
Matrix Proteins ◽  
Crystal Formation ◽  
Extracellular Matrix Components ◽  
Dentin Mineralization ◽  
Matrix Maturation ◽  
Cell Matrix Interactions

The extracellular matrix of dentin consists of mineral (hydroxyapatite), collagen, and several noncollagenous matrix proteins. These noncollagenous matrix proteins may be mediators of cell-matrix interactions, matrix maturation, and mineralization. This review describes the current knowledge of the chemistry of mineral crystal formation in dentin with special emphasis on the roles of the dentin matrix proteins. The functions of some of these matrix proteins in the mineralization process have been deduced based on in vitro studies. Functions for others have been postulated based on analogy with some of the bone matrix proteins. Evidence suggests that several of these matrix proteins may have multiple effects on nucleation, crystal growth, and orientation of dentin hydroxyapatite.

Magnesium Reduces Calcium Oxalate Crystal Formation in Human Whole Urine

1982 ◽  
Vol 62 (1) ◽  
pp. 17-19 ◽  
Author(s):  
P. C. Hallson ◽  
G. A. Rose ◽  
S. Sulaiman
Keyword(s):  
Calcium Oxalate ◽  
Inverse Correlation ◽  
Normal Volunteer ◽  
Urine Samples ◽  
Magnesium Concentration ◽  
Crystal Formation ◽  
Calcium Oxalate Crystal ◽  
Isotopic Methods ◽  
Whole Urine

1. A low urinary magnesium was induced in normal volunteer subjects by giving cellulose phosphate; magnesium was added in vitro to yield urine samples of normal and high magnesium concentrations 2. After rapid evaporation of these urine samples at pH 5.3 to standard osmolality the calcium oxalate crystals were measured by microscopy and isotopic methods 3. There was a clear inverse correlation between magnesium concentration and calcium oxalate crystal formation 4. The case for treating calcium oxalate urolithiasis with magnesium is strengthened.

Low-vitamin E diet exacerbates calcium oxalate crystal formation via enhanced oxidative stress in rat hyperoxaluric kidney

2009 ◽  
Vol 296 (1) ◽  
pp. F34-F45 ◽  
Author(s):  
Ho-Shiang Huang ◽  
Ming-Chieh Ma ◽  
Jun Chen
Keyword(s):  
Oxidative Stress ◽  
Drinking Water ◽  
Vitamin E ◽  
Calcium Oxalate ◽  
Oxidative Enzyme ◽  
Separate Experiment ◽  
Crystal Formation ◽  
Standard Diet ◽  
Crystal Deposition ◽  
Caox Crystal

Vitamin E was previously reported to reduce calcium oxalate (CaOx) crystal formation. This study explored whether vitamin E deficiency affects intrarenal oxidative stress and accelerates crystal deposition in hyperoxaluria. The control (C) group of rats received a standard diet and drinking water, while the experimental groups received 0.75% ethylene glycol (EG) in drinking water for 42 days. Of the latter, one group received a standard diet (EG group), one received a low-vitamin E (LE) diet (EG+LE group), and the last received an LE diet with vitamin E supplement (4 mg) (EG+LE+E group). The C+LE and C+LE+E groups were the specific controls for the last two experimental groups, respectively. In a separate experiment, EG and EG+LE rats were studied on days 3–42 to examine the temporal relationship between oxidative change and crystal formation. Urinary biochemistry and activity/levels of antioxidative and oxidative enzymes in glomeruli and tubulointerstitial specimens (TIS) were examined. In EG rats, CaOx crystal accumulation was associated with low antioxidative enzyme activity in TIS and with increased oxidative enzyme expression in glomeruli. In the EG+LE group, marked changes in antioxidative and oxidative enzyme levels were seen and correlated with massive CaOx deposition and tubular damage. The increased oxidative stress seen with EG+LE treatment was largely reversed by vitamin E supplementation. A temporal study showed that decrease in antioxidative defense and increased free radical formation in the EG+LE group occurred before crystal deposition. This study shows that low vitamin E disrupts the redox balance and causes cell death, thereby favoring crystal formation.

In vitro and clinical study of oxalate influence on calcium oxalate crystal formation

1988 ◽  
Vol 87 (4) ◽  
pp. 494-506 ◽  
Author(s):  
Y. Berland ◽  
M. Olmer ◽  
M. Grandvuillemin ◽  
H.E. Lundager Madsen ◽  
R. Boistelle
Keyword(s):  
Clinical Study ◽  
Calcium Oxalate ◽  
Crystal Formation ◽  
Calcium Oxalate Crystal

scholarly journals Lactiplantibacillus plantarum Reduced Renal Calcium Oxalate Stones by Regulating Arginine Metabolism in Gut Microbiota

2021 ◽  
Vol 12 ◽  
Author(s):  
Yu Liu ◽  
Xi Jin ◽  
Lei Tian ◽  
Zhongyu Jian ◽  
Yucheng Ma ◽  
...  
Keyword(s):  
Drinking Water ◽  
Gut Microbiota ◽  
Calcium Oxalate ◽  
Control Group ◽  
Arginine Metabolism ◽  
Model Group ◽  
Crystal Deposition ◽  
Caox Crystal ◽  
Relative Abundances

Renal calcium oxalate (CaOx) stones are a common kidney disease. There are few methods for reducing the formation of these stones. However, the potential of probiotics for reducing renal stones has received increasing interest. We previously isolated a strain of Lactiplantibacillus plantarum N-1 from traditional cheese in China. This study aimed to investigate the effects of N-1 on renal CaOx crystal deposition. Thirty rats were randomly allocated to three groups: control group (ddH2O by gavage), model group [ddH2O by gavage and 1% ethylene glycol (EG) in drinking water], and Lactiplantibacillus group (N-1 by gavage and 1% EG in drinking water). After 4 weeks, compared with the model group, the group treated with N-1 exhibited significantly reduced renal crystals (P &lt; 0.05). In the ileum and caecum, the relative abundances of Lactobacillus and Eubacterium ventriosum were higher in the control group, and those of Ruminococcaceae UCG 007 and Rikenellaceae RC9 were higher in the N-1-supplemented group. In contrast, the relative abundances of Staphylococcus, Corynebacterium 1, Jeotgalicoccus, Psychrobacter, and Aerococcus were higher in the model group. We also predicted that the arginase level would be higher in the ileal microbiota of the model group than in the N-1-supplemented group with PICRUSt2. The arginase activity was higher, while the level of arginine was lower in the ileal contents of the model group than in the N-1-supplemented group. The arginine level in the blood was also higher in the N-1-supplemented group than in the model group. In vitro studies showed that exposure to arginine could reduce CaOx crystal adhesion to renal epithelial HK-2 cells. Our findings highlighted the important role of N-1 in reducing renal CaOx crystals by regulating arginine metabolism in the gut microbiota. Probiotics containing L. plantarum N-1 may be potential therapies for preventing renal CaOx stones.

Strain differences in urinary factors that promote calcium oxalate crystal formation in the kidneys of ethylene glycol-treated rats

2009 ◽  
Vol 296 (5) ◽  
pp. F1080-F1087 ◽  
Author(s):  
Yan Li ◽  
Kenneth E. McMartin
Keyword(s):  
Ethylene Glycol ◽  
Calcium Oxalate ◽  
Wistar Rats ◽  
Kidney Tissue ◽  
Wistar Rat ◽  
Crystal Formation ◽  
Rat Strains ◽  
Crystal Deposition ◽  
Caox Crystal ◽  
F344 Rats

Ethylene glycol (EG)-induced hyperoxaluria is the most commonly employed experimental regimen as an animal model of calcium oxalate (CaOx) stone formation. The variant sensitivity to CaOx among different rat strains has not been fully explored, although the Wistar rat is known to accumulate more CaOx in kidney tissue after low-dose EG exposure than in the Fischer 344 (F344) rats. Supersaturation of CaOx in tubular fluid contributes to the amount of CaOx crystal formation in the kidney. We hypothesized that the urinary supersaturation of CaOx in Wistar rats is higher than that of F344 rats, thereby allowing for greater CaOx crystal deposition in the Wistar rat. Age-matched male Wistar and F344 rats were treated with 0.75% EG or drinking water for 8 wk. Twenty-four-hour urine was collected at 0, 2, 4, 6, and 8 wk for analysis of key electrolytes to calculate the CaOx supersaturation. Plasma oxalate level was also measured. Our data confirmed the different sensitivity to renal toxicity from EG between the two rat strains (Wistar > F344). After EG treatment, the plasma oxalate level and urine oxalate excretion were markedly greater in the Wistar rats than in the F344 rats, while urine calcium was slightly decreased in Wistars. Thus, the CaOx supersaturation in urine of Wistar rats was higher, which led to a greater crystal deposition in kidney in Wistar rats. These studies suggest that during EG treatment, changes in urine electrolytes and in CaOx supersaturation occur to a greater extent in the Wistar rat, in agreement with its greater sensitivity to EG toxicity.

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