Encapsulation of Leflunomide (LFD) in a novel niosomal formulation facilitated its delivery to THP-1 monocytic cells and enhanced Aryl hydrocarbon receptor (AhR) nuclear translocation and activation
Leflunomide (LFD) is an Aryl hydrocarbon receptor (AhR) agonist and immunomodulatory drug with several side effects. Niosomes are novel drug delivery systems used to reduce the unfavorable effects of drugs by enhancing their bioavailability, controlling their release and targeting specific sites.
Here, we prepared niosomal formulations of LFD, evaluated their properties and delivered to THP-1 monocytic cells to study the activation and nuclear translocation of AhR.
Four types of non-ionic surfactants were utilized to formulate niosomes by thin film hydration (TFH) method. Entrapment efficiency (EE %) of niosomes were quantified and dynamic light scattering (DLS) was performed. Transmission electron microscopy (TEM) was used to identify the morphology of LFD niosomes. Dialysis method was used to measure LFD release rate. MTS assay was adopted to examine the viability of the cells upon each treatment. The nuclear transfer of AhR was investigated by Immunocytochemistry (ICC). The mRNA expression of IL1β and CYP1A1 were evaluated using quantitative RT-PCR.
Span 60: cholesterol (1:1) showed the highest EE% (70.00 ± 6.24), largest particles (419.00 ± 4.16 nm) and the best uniformity with the lowest PDI (0.291 ± 0.007). TEM micrographs of Span 60 (1:1) nanoparticles showed conventional spherical vesicles with internal aqueous spaces. The release rate of LFD from Span 60 (1:1) vesicles was slower. Although the viability of LFD niosome-treated THP-1 cells was decreased, they were associated with lower cytotoxic effects compared with the free LFD counterparts. Both free and niosomal LFD treatments intensified the nuclear translocation of AhR. The mRNA expression of CYP1A1 was overexpressed while IL1β was downregulated in both free and niosomal LFD treated combinations.
LFD encapsulation in Span 60: cholesterol (1:1) niosomal formulation could be introduced as a suitable vehicle of transferring LFD to THP-1 cells, with minimal cytotoxic effects, enhancing the AhR nuclear translocation and activation and inducing immunomodulatory properties.
KeywordsAryl hydrocarbon receptor (AhR) Drug delivery Leflunomide (LFD) Niosome
This article was derived from a thesis of M.Sc. degree in the field of Medical biotechnology (Grant Number: 960129002) at Gorgan School of Advanced Technologies in Medicine of Golestan University of Medical Sciences, Gorgan, Iran. We would like to thank Dr. Samadian, Mrs. Yousefi and Mrs. Haydari for their scientific and technical support.
MH: Acquisition of data, Analyses and interpretations of data, Manuscript drafting, Revision of the manuscript. NAS: Participation in data acquisition, analyses and interpretations. MSA: Participation in data acquisition. BK: Participation in data acquisition. SM: Study design and concept, participation in literature bibliography, data acquisition and analysis, manuscript drafting and critical revision of the manuscript. YY: Participation in study design and final revision of the manuscript. All authors read and approved the final version of the manuscript.
Compliance with ethical standards
Conflict of interests
On behalf of all authors, the corresponding author states that there is no conflict of interest.
Consent for publication
Ethics approval and consent to participate
The study was approved by the Ethics committee of Golestan University of Medical Sciences (Code of Ethics: IR.GUOMS.REC.13950259).
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