Evaluation of the Cytotoxicity of α-Cyclodextrin Derivatives on the Caco-2 Cell Line and Human Erythrocytes.

Eszter Róka,Florent Perret,Ferenc Fenyvesi,Judit Váradi,Ildikó Bácskay,Rudolf Gesztelyi,Pálma Fehér,Éva Fenyvesi,Lajos Szente,Caroline Félix,Alexandra Bocsik,Mária Deli,Zoltán Ujhelyi,Miklós Vecsernyés

doi:10.3390/molecules201119694

Abstract

Cyclodextrins, even the 6-membered α-cyclodextrin, are approved in the various pharmacopoeias as pharmaceutical excipients for solubilizing and stabilizing drugs as well as for controlling drug release. Recently α-cyclodextrin has also been marketed as health food with beneficial effects on blood lipid profiles. However, the concentration of α-cyclodextrin used may be very high in these cases, and its toxic attributes have to be seriously considered. The objective of this study was to investigate the cytotoxicity of various, differently substituted α-cyclodextrin derivatives and determine relationship between the structures and cytotoxicity. Three different methods were used, viability tests (MTT assay and Real Time Cell Electronic Sensing on Caco-2 cells) as well as hemolysis test on human red blood cells. The effect of α-cyclodextrin derivatives resulted in concentration-dependent cytotoxicity, so the IC50 values have been determined. Based on our evaluation, the Real Time Cell Electronic Sensing method is the most accurate for describing the time and concentration dependency of the observed toxic effects. Regarding the cytotoxicity on Caco-2 cells, phosphatidylcholine extraction may play a main role in the mechanism. Our results should provide help in selecting those α-cyclodextrin derivatives which have the potential of being used safely in medical formulations.

Highlights

Cyclodextrins (CDs) are widely used excipients and still in the focus of drug development [1]
The resulting data are biologically relevant because the elimination of labels brings the cells closer to physiological conditions. It has been confirmed by the study of Ozsvári et al [34] who observed similar results of cytotoxicity using immortalized and rat primary cell lines in real time cell electronic sensing assay (RT-CES) experiments
The derivatives used were: phosphated α-CD sodium salt, carboxymethylated α-CD sodium salt (CMACD), sulfated α-CD carboxymethylated α-CD polymer crosslinked with epichlorohydrin (CMACDEp), randomly methylated α-CD (RAMEA), acetylated α-CD (AcACD), hexakis-(2,3,6-tri-O-methyl)-α-CD (TRIMEA), (2-hydroxy) propyl α-CD (HPACD) and succinylated α-CD (SuACD)

Summary

Introduction

Cyclodextrins (CDs) are widely used excipients and still in the focus of drug development [1] They are used as solubilizing factors, protective agents for light-sensitive drugs, and as a part of sustained release or drug delivery systems [2]. In some cases their stabilizing effect against hydrolysis, oxidation, and microbial decomposition is utilized. There are several products on the market (tablets, eye drops, transdermal patches, inhalers, etc.) which contain various CDs already approved as pharmaceutical ingredients [3,4] Most of these products contain β-CD or its hydroxypropyl, and sulfobutyl derivatives, and only a few of the marketed formulations are produced with α-CD. Α-CD like β-CD is practically not absorbed from the gastrointestinal tract explaining the low oral toxicity, it might damage the cells of the intestine, especially at the high applied concentrations used in these applications

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Conclusion