Cross-linked chitosan/liposome hybrid system for the intestinal delivery of quercetin

doi:10.1016/j.jcis.2015.09.013

Journal of Colloid and Interface Science

Volume 461, 1 January 2016, Pages 69-78

https://doi.org/10.1016/j.jcis.2015.09.013 Get rights and content

Abstract

Quercetin is a flavonoid with antioxidant/anti-inflammatory properties, poorly absorbed when administered orally. To increase its bioavailability and optimize its release in the intestine, a hybrid system made of liposomes coated with cross-linked chitosan, named TPP-chitosomes, was developed and characterized by light scattering, transmission electron microscopy, differential scanning calorimetry, X-ray powder diffraction and Turbiscan® technology. The TPP-chitosomes were nanosized (∼180 nm), fairly spherical in shape and unilamellar. The actual coating of the surface of liposomes with the cross-linked chitosan was demonstrated by Small-Angle X-ray Scattering.

The release of quercetin in simulated gastric and intestinal pH was investigated, the results showing that the system provided resistance to acidic conditions, and promoted the release in alkaline pH, mimicking the intestinal environment.

The proposed hybrid system represents a promising combination of nanovesicles and chitosan for the delivery of quercetin to the intestine in the therapy of oxidative stress/inflammation related disorders.

Graphical abstract

Introduction

Quercetin (3,3′,4′,5,7-pentahydroxyflavone) is a flavonoid present in a large number of edible vegetables and fruits [1], [2]. This molecule has many health-promoting effects, including improvement of cardiovascular health, reducing risk for cancer, and coping with inflammatory disorders, mainly related to its strong antioxidant action, by which it upregulates the endogenous free radical defenses [3], [4]. However, the efficacy of quercetin is limited due to its hydrophobicity, instability in physiological media, poor gastrointestinal absorption, and extensive xenobiotic metabolism at intestines and liver (i.e., glucuronidation or sulfation), which collectively contribute to its low oral bioavailability [4], [5], [6], [7]. Various nanotechnological approaches have been used to enhance its solubility, dissolution rate, and hence, bioavailability, including solid dispersions, nanosuspensions, microemulsions, solid lipid nanoparticles and prodrugs [8], [9], [10], [11], [12]. Therefore, a critical need exists to develop alternative formulative strategies to overcome the shortcomings of quercetin and enhance its bioavailability.

The objective of this study was to develop, optimize and thoroughly characterize a formulation that enhances quercetin local bioavailability, bypasses the stomach and allows its release in the intestine for the treatment of disorders in which oxidative stress and inflammation are involved [2], [13], [14]. Among the different approaches to achieve intestinal drug delivery, the use of natural polymers holds promise. In recent years, polymer-coated liposomes have been proposed for the targeted delivery of drugs to the inflamed intestinal mucosa, upon oral administration [14]. Conventional liposomes can be hardly used, due to their low resistance to gastric pH and enzymatic degradation, but they can be easily protected by a polymeric coating. Chitosan is one of the most widely used polymers for coating liposomes. It is a linear cationic polysaccharide comprised of (1 → 4)-linked units of glucosamine and N-acetyl-glucosamine, mainly produced by partial deacetylation of chitin. Due to its unique and attractive properties, including biocompatibility, biodegradability, biorenewability and bioadhesion, chitosan is widely explored for biopharmaceutical applications, especially for intestinal drug delivery [15], [16]. However, it has a limited capacity for controlling drug release from oral dosage forms due to its fast dissolution in the stomach. To overcome this disadvantage, three dimensional networks have been developed by non-covalent complexation relying on electrostatic, hydrophobic and/or hydrogen bonding forces. The cationic amino groups of the repeating glucosamine units of chitosan can interact electrostatically with the anionic groups of other polyions to form polyelectrolyte complexes. These cross-linking interactions are physical in nature and reversible, but can provide the required properties for optimal drug delivery if the polymer is properly complexed. In a previous work, chitosan was complexed with Nutriose FM06®, a branched dextrin obtained from starch, and used to coat polyethylene glycol-containing vesicles, with the aim of producing an enzyme-sensitive, prebiotic, delayed release system for the delivery of quercetin to inflamed colon [14].

In the present paper, chitosan was complexed with sodium tripolyphosphate and used to coat liposomes, thus obtaining a hybrid system conceived for increasing the bioavailability of incorporated quercetin and optimizing its release rate to reach the intestine. To achieve these goals, it was imperative to assess the effectiveness and feasibility of the system by a thorough characterization using different analytical tools, such as Light Scattering, Transmission Electron Microscopy (TEM), Differential Scanning Calorimetry (DSC), X-ray Powder Diffraction (XRPD), and Small-Angle X-ray Scattering (SAXS). Further, the ability of the system to release quercetin in simulated gastric and intestinal pH was investigated.

Section snippets

Materials

Phospholipon90H and Phospholipon50 (P90H and P50; phosphatidylcholine-based phospholipids) were purchased from Lipoid GmbH (Ludwigshafen, Germany). Cholesterol (CHOL), chitosan of low molecular weight (50–190 kDa; degree of deacetylation ⩾75%), sodium tripolyphosphate (TPP), phosphate buffered saline (PBS, pH 7.4) and quercetin (QUE) were purchased from Sigma–Aldrich (Milan, Italy).

Preparation of TPP-chitosan/liposome hybrid system

P90H (60 mg/ml), P50 (30 mg/ml), CHOL (2 mg/ml) and quercetin (5 mg/ml) were weighed in a glass vial, PBS was added and

Hybrid system’s fabrication and key features

A hybrid system made of quercetin-loaded liposomes coated with TPP cross-linked chitosan was developed aiming at realizing an efficient intestinal delivery system by exploiting the attractive biopharmaceutical properties of the flavonoid, the phospholipid vesicles, and the polymer, as well.

The polycationic chitosan was dissolved in acetic acid solution to protonate the free amino groups and allow the supramolecular self-organizing interaction with negatively charged liposomes on the one hand,

Conclusions

The goal of this work was to design a formulation for the intestinal delivery of quercetin, with great potential for health promotion. The development of such system that achieves both therapeutic effectiveness and biocompatibility is a challenging task, since quercetin has low oral bioavailability. In this context, a cross-linked chitosan/liposome hybrid nanosystem loading quercetin was successfully fabricated, without any complicated apparatus or organic solvents. The morphology and structure

Acknowledgement

Dr. Pons acknowledges financial support from MINECO-CTQ2013-41514-P.

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