Abstract: SA-PO309
Calcium Citrate Incorporated into Calcium Carbonate and Calcium Carbonate Nanoparticles Alleviate Cellular Injury from Acidosis
Session Information
- Fluid and Electrolytes: Basic - II
November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Fluid and Electrolytes
- 901 Fluid and Electrolytes: Basic
Authors
- Dissayabutra, Thasinas, Chulalongkorn University, Pathumwan, Bangkok, Thailand
- Udomsinsirikul, Tipagorn, NSTDA, Bangkok, Thailand
- Rojanathanes, Rojrit, Chulalongkorn University, Pathumwan, Bangkok, Thailand
- Sereemaspun, Amornpun, Chulalongkorn University, Pathumwan, Bangkok, Thailand
Group or Team Name
- Renal Biochemistry and Stone Disease Research Unit, Faculty of Medicine, Chulalongkorn University
Background
Metabolic acidosis is a common complication found in patients with chronic kidney disease (CKD), and causes cellular dysfunctions, protein degradation, inflammation, oxidative stress and cell death. Current medication to treat metabolic acidosis is the supplementation of alkalinizing agents such as sodium bicarbonate and sodium citrate. We proposed new nanoparticles as the adjuvant therapy to mitigate the consequences of metabolic acidosis in CKD patients.
Methods
Calcium citrate incorporated into calcium carbonate nanoparticles (CCNP) and calcium carbonate nanoparticles (CNP) were synthesized from calcium chloride and sodium citrate. A HK-2 cells cultured in DMEM with pH 7.4 (normal environment) and 4.0-5.0 (acidosis environment) were used to study the cellular toxicity by Resazurin oxidative-reduction assay, intracellular reactive oxygen species (ROS) production by 2,7-dichlofluorescein-diacetate (DCFH-DA) test, and cell death by flow cytometry. Fluorescence isothiocyanate (FITC) conjugated CCNP and CNP were used to study cellular uptake. Intracellular and extracellular bicarbonate concentration were measured by automated biochemical analyzer. Sodium citrate (Nacit) was used as a standard drug in these experiments.
Results
CNP and CCNP had very low cytotoxicity at the concentration up to 1 mg/ml. The results showed that CNP and CCNP did not alter the extracellular pH, or extracellular and intracellular bicarbonate concentration, and they were freely uptake into the cell in normal and acidic condition. Pharmacological tests revealed that both CNP and CCNP can suppress ROS production better than sodium citrate treatment. In addition, CNP and CCNP treatment ameliorated acidosis-induced cell death and apoptosis.
Conclusion
CNP and CCNP mitigates the consequences of metabolic acidosis, as they could reduce ROS production and cell death in acidic environment. Because they did not alter pH and bicarbonate concentration, they could be potentially used as the adjuvant therapy with alkalinizing agents to treat metabolic acidosis in CKD patients. A study of in vivo effects of these nanoparticles is ongoing.
Funding
- Government Support - Non-U.S.