Biochemical Properties of Alcohol Dehydrogenase and Glutamate Dehydrogenase Encapsulated into Human Erythrocytes by a Hypotonic-Dialysis Procedure

Sanz, Sílvia; Lizano, Carmen; Garín, Marina I.; Luque, J.; Pinilla, Montserrat

doi:10.1007/978-1-4899-0044-9_14

Cited by 8 publications

(8 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…EBRs for removal of low molecular metabolites (ethanol, methanol, cyanide, glucose or ammonium) from bloodstream have been described. These EBRs were based on alcohol dehydrogenase [11,59,60], alcohol oxidase [61], acetaldehyde dehydrogenase [11,62] or alcohol-and acetaldehyde dehydrogenase together [63] in cases of ethanol, methanol and acetaldehyde removal.…”

Section: Erythrocytes-bioreactors For Low Molecular Metabolites Utilimentioning

confidence: 99%

“…EBRs for ammonium removal (so-called ammocytes) have been developed by various scientific groups. For enapsulation into RBCs, glutamate dehydrogenase was used, which catalyzed the formation of l-glutamic acid from α-ketoglutarate and ammonium in the presence of NADPH [59,60,72], as well as glutamine synthetase, which catalyzed the formation of l-glutamine from l-glutamic acid and ammonium in the presence of ATP [73,74]. Each of these enzymes was encapsulated into RBCs using reversible hypoosmotic dialysis.…”

Section: Ammocytesmentioning

confidence: 99%

“…Sometimes, the work of the enzyme inside the RBCs may be limited by the rate of transport for the necessary substrate of the reaction into the cells. This happened, for example, when ammocytes based on glutamate dehydrogenase [59,60,72] or glutamine synthetase [73,74] were created. In this case, the researchers proposed for incorporation into the RBCs a new enzyme system consisting of two enzymes that provided cyclic consumption and production of the necessary metabolites inside the cell.…”

Section: Limitations Of the Rbcs' Use As Drug Carriersmentioning

confidence: 99%

“…β-Galactosidase - [21] β-Glucocerebrosidase (β-glucosidase) Gaucher disease [21,37,38,[44][45][46]250] β-Glucuronidase Syndrome Slaya [251] l-Phenylalanine ammonia lyase Phenylketonuria [48,252,253] Phenylalanine hydroxylase [50,254] Uricase (uratoxidase) Uric acid removal [255,256] Urease, urease + alanine dehydrogenase Urea utilization [257][258][259] Adenosine deaminase Severe combined immunodeficiency caused by deaminase deficiency [27,[55][56][57][58]260] Thymidine phosphorylase Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) [56,246,261,262] Glutamate dehydrogenase Hyperammonemia [59,60,72] Glutamine synthetase [73,74] Glutamate dehydrogenase + alanine aminotransferase [11,63] Arginase Hyperammonemia due to arginase deficiency [263] Alcohol dehydrogenase…”

Section: Active Substance Application Referencesmentioning

confidence: 99%

“…Alcohol and methanol intoxication [59,60] Alcohol oxidase [61] Acetaldehyde dehydrogenase [62] Alcohol dehydrogenase + acetaldehyde dehydrogenase [11,63]…”

Section: Active Substance Application Referencesmentioning

confidence: 99%

See 4 more Smart Citations

Erythrocytes as Carriers: From Drug Delivery to Biosensors

et al. 2020

View full text Add to dashboard Cite

Drug delivery using natural biological carriers, especially erythrocytes, is a rapidly developing field. Such erythrocytes can act as carriers that prolong the drug's action due to its gradual release from the carrier; as bioreactors with encapsulated enzymes performing the necessary reactions, while remaining inaccessible to the immune system and plasma proteases; or as a tool for targeted drug delivery to target organs, primarily to cells of the reticuloendothelial system, liver and spleen. To date, erythrocytes have been studied as carriers for a wide range of drugs, such as enzymes, antibiotics, anti-inflammatory, antiviral drugs, etc., and for diagnostic purposes (e.g., magnetic resonance imaging). The review focuses only on drugs loaded inside erythrocytes, defines the main lines of research for erythrocytes with bioactive substances, as well as the advantages and limitations of their application. Particular attention is paid to in vivo studies, opening-up the potential for the clinical use of drugs encapsulated into erythrocytes.Pharmaceutics 2020, 12, 276 2 of 44 property of RBCs allows to load them with biologically active substances of different molecular weights. For these reasons, erythrocytes are promising biocompatible cells for drug delivery.The methods for incorporating various substances into red blood cells differ in the way that substances penetrate the cells. The cause of permeability may be the pores' formation in the cell membrane due to a physical exposure (high voltage electric pulse [10,11] or ultrasound [12]). Drug molecules can also enter the RBCs by endocytosis in the presence of certain chemical compounds (for example, primaquine [13], vinblastine, chlorpromazine, hydrocortisone or tetracaine [14,15]), or using the cell-penetrating peptides bounded to the compound that should be encapsulated [16]. However, the most popular are different variants of osmotic methods.In some cases, RBCs are first exposed to a hyperosmotic pulse of a low molecular weight substance that penetrates very well through the cell membrane (for example, dimethyl sulfoxide (DMSO) [17,18] or glucose [19,20]). After washing the cells, which decreases the external concentration of these compounds and creates a gradient of their concentration between both sides of the RBC membrane, the target drug is introduced into the external volume. Water with this drug begins to enter into the cells to decrease the osmotic pressure there. The process ends when the gradient of DMSO or glucose disappears. The pores close and part of the drug remains into RBCs. Other, the most popular of the osmotic methods are hypoosmotic. These methods are based on creating a hypotonic environment around RBCs, which causes swelling of the cells and opening pores in the cellular membrane, through which therapeutic compounds can penetrate RBCs. Then, a hypertonic solution is introduced into the cell suspension. The pores close, the cells restore their original size, trapping the drug molecules inside the cell. Osmotic methods are divided into severa...

show abstract

Section: Erythrocytes-bioreactors For Low Molecular Metabolites Utilimentioning

confidence: 99%

Section: Ammocytesmentioning

confidence: 99%

Section: Limitations Of the Rbcs' Use As Drug Carriersmentioning

confidence: 99%

Section: Active Substance Application Referencesmentioning

confidence: 99%

“…Alcohol and methanol intoxication [59,60] Alcohol oxidase [61] Acetaldehyde dehydrogenase [62] Alcohol dehydrogenase + acetaldehyde dehydrogenase [11,63]…”

Section: Active Substance Application Referencesmentioning

confidence: 99%

See 3 more Smart Citations

Erythrocytes as Carriers: From Drug Delivery to Biosensors

et al. 2020

View full text Add to dashboard Cite

show abstract

In vitro study of alcohol dehydrogenase and acetaldehyde dehydrogenase encapsulated into human erythrocytes by an electroporation procedure

Lizano

Sanz

Luque

et al. 1998

Biochimica et Biophysica Acta (BBA) - General Subjects

View full text Add to dashboard Cite

Erythrocytes as bioreactors to decrease excess ammonium concentration in blood

Protasov

Борсакова

Alexandrovich

et al. 2019

Sci Rep

View full text Add to dashboard Cite

Increased blood ammonium concentrations cause neurological complications. Existing drugs are not always sufficiently effective. Alternatively, erythrocytes-bioreactors (EBRs) loaded with enzymes utilizing ammonium, were suggested for ammonium removal from blood. However all they worked only for a short period of time. The reasons for this were not investigated. In this study, EBR mathematical models were developed and analysed based on the reactions of glycolysis and different enzymes utilizing ammonium, which showed that the efficiency and duration of EBRs’ functioning could be limited due to low permeability of the cell membrane for some key substrates and products. A new enzyme system including glutamate dehydrogenase and alanine aminotransferase was proposed and realised experimentally, which was not limited by cell membrane permeability for glutamate and α-ketoglutarate due to creating metabolic pathway where these metabolites were produced and consumed cyclically. New bioreactors removed ammonium in vitro at the rate of 1.5 mmol/h × lRBCs (for human bioreactors) and in vivo in a model of hyperammoniemia in mice at the rate of 2.0 mmol/h × lRBCs (for mouse bioreactors), which correlated with model calculations. Experimental studies proved the proposed mathematical models are correct. Mathematical simulation of erythrocyte-bioreactors opens new opportunities for analysing the efficiency of any enzyme included in erythrocytes.

show abstract

Biochemical Properties of Alcohol Dehydrogenase and Glutamate Dehydrogenase Encapsulated into Human Erythrocytes by a Hypotonic-Dialysis Procedure

Cited by 8 publications

References 12 publications

Erythrocytes as Carriers: From Drug Delivery to Biosensors

Erythrocytes as Carriers: From Drug Delivery to Biosensors

In vitro study of alcohol dehydrogenase and acetaldehyde dehydrogenase encapsulated into human erythrocytes by an electroporation procedure

Erythrocytes as bioreactors to decrease excess ammonium concentration in blood

Contact Info

Product

Resources

About