Loading Mg²⁺ into Erythrocytes with a Proposed Single Syringe Sterile Loading Process

Abstract

Primary hypomagnesemia with secondary hypocalcemia (PHSH) is a genetically defined disease in which there is a malabsorption of magnesium from the intestine. The aim of this research was to encapsulate magnesium into autologous erythrocytes to be used in PHSH patients. A scheme to use a single syringe closed sterile system for the process is proposed. Controlled lysis of erythrocytes and the ability to reseal the loaded erythrocytes were investigated. Method: Suitable concentrations of salt solutions required for making reversible pores and resealing of erythrocytes were identified. For making reversible pores, 0.7% NaCl and 2.3% MgSO 4 were used separately. A solution of 3.5% MgSO 4 was used as the drug loading solution. After the loading step, the isotonicity for the erythrocytes was restored with 0.9% NaCl and 3.0% MgSO 4 solutions separately. The ability to load and reseal erythrocytes were assessed by determining the resulting Mg 2+ concentration through Microwave Plasma Atomic Emission Spectrophotometric (MP-AES) analysis. Results: Analysis of results revealed that Mg 2+ ions can be successfully loaded into erythrocytes through the formation of reversible pores. Resealing failed with 3.0% MgSO 4 , but the erythrocytes were successfully resealed with 0.9% NaCl solution. Conclusion: The need for strictly controlled salt solution concentrations for lysis and resealing were identified. In theoretical concentration determinations, the osmotic coefficient had to be taken into account. The study showed the possibility of loading Mg 2+ into erythrocytes as a successful drug delivery system.