Efficient Immuno-Modulation of TH1/TH2 Biomarkers in 2,4-Dinitrofluorobenzene-Induced Atopic Dermatitis: Nanocarrier-Mediated Transcutaneous Co-Delivery of Anti-Inflammatory and Antioxidant Drugs

Efficient Immuno-Modulation of TH1/TH2 Biomarkers in 2,4-Dinitrofluorobenzene-Induced Atopic Dermatitis: Nanocarrier-Mediated Transcutaneous Co-Delivery of Anti-Inflammatory and Antioxidant Drugs Zahid Hussain, Haliza Katas*, Mohd Cairul Iqbal Mohd Amin, Endang Kumolosasi Centre for Drug Delivery Research, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia Abstract The present study was conducted with the aim to investigate the immuno-modulatory and histological stabilization effects of nanocarrier–based transcutaneous co-delivery of hydrocortisone (HC) and hydroxytyrosol (HT). In this investigation, the clinical and pharmacological efficacies of nanoparticle (NP)-based formulation to alleviate 2,4-dinitrofluorobenzene (DNFB)induced atopic dermatitis (AD) was explored by using an NC/Nga mouse model. Ex vivo visual examination of AD induction in experimental mice indicated remarkable control of NP-based formulations in reducing pathological severity of AD-like skin lesions. Therapeutic effectiveness of NP-based formulations was also evaluated by comparing skin thickness of ADinduced NP-treated mice (456627 mm) with that of atopic mice (916637 mm). Analysis of the immuno-spectrum of AD also revealed the dominance of NP-based formulations in restraining immunoglobulin-E (IgE), histamine, prostaglandin-E2 (PGE2), vascular endothelial growth factor-a (VEGF-a), and T-helper cells (TH1/TH2) producing cytokines in serum and skin biopsies of tested mice. These anti-AD data were further supported by histological findings that revealed alleviated pathological features, including collagen fiber deposition, fibroblasts infiltration, and fragmentation of elastic fibers in experimental mice. Thus, NP-mediated transcutaneous co-delivery of HC and HT can be considered as a promising therapy for managing immunological and histological spectra associated with AD. Citation: Hussain Z, Katas H, Mohd Amin MCI, Kumolosasi E (2014) Efficient Immuno-Modulation of TH1/TH2 Biomarkers in 2,4-Dinitrofluorobenzene-Induced Atopic Dermatitis: Nanocarrier-Mediated Transcutaneous Co-Delivery of Anti-Inflammatory and Antioxidant Drugs. PLoS ONE 9(11): e113143. doi:10.1371/journal. pone.0113143 Editor: Hiroshi Shiku, Mie University Graduate School of Medicine, Japan Received May 19, 2014; Accepted October 23, 2014; Published November 14, 2014 Copyright: ß 2014 Hussain et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability: The authors confirm that all data underlying the findings are fully available without restriction. All relevant data are within the paper. Funding: This work was supported by the UKM Arus Perdana grant (AP-2013-002). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript Competing Interests: The authors have declared that no competing interests exist. * Email: a), a potent biomarker that induces hyperpermeability of blood vessels via abnormal neovascularization and endothelial cell proliferation. VEGF-a also acts as a chemoattractant for various inflammatory cells responsible for persistent aggravation in erythema and edema [7,8]. In addition, release of numerous TH1/TH2-specific inflammatory mediators, such as interleukin (IL) types IL-4, IL-5, IL-6, IL-12p70, IL-13, interferon-c (IFN-c) and tumor necrosis factor-a (TNF-a) has been demonstrated in patients with AD [9,10]. Topical glucocorticoids (TGs) are recognized as a wellestablished mainstay in relieving acute and chronic exacerbation of psoriasis and AD [11,12]. The clinical significance of TGs in the prevention of these inflammatory disorders is concurrent with their vasoconstrictive, anti-inflammatory, immunosuppressive, and antiproliferative potency. However, long-term use of TGs is often accompanied by several local and systemic deleterious effects [13,14] that limit clinical significance and exclude their application in chronic maintenance therapies. Hence, hydrocortisone (HC), a mildly potent agent of TGs, is administered percutaneously to minimize unwanted effects associated with use of TGs [3,12]. In addition, HC is recognized as a mild agent due to its minimal Introduction Atopic dermatitis (AD) is chronically relapsing, non-contagious, and exudative; it typically manifests as pruritic dermatosis accompanied by perivascular infiltration of T-helper (TH1/TH2)lymphocytes, mast cells, and immunoglobulin-E (IgE) [1,2]. Common signs and symptoms of AD include the appearance of red to brownish-grey colored patches, severe itching, small raised bumps with exudates/transudates, and cracked/damaged stratum corneum (SC) [3,4]. Genetic variability, environmental interactions, skin barrier disorders, and immunological reactions are among the proposed contributing factors [5,6]; however, the exact pathogenesis of this allergic disorder is not well-established yet. Mast cells and basophils are among the key effector cells in IgEmediated allergic disorders, and play a key role in the pathogenesis of AD. These cells are stimulated in response to active crosslinking of AD-specific IgE with high affinity cell-surface IgEreceptors. On activation, these cells endure degranulation. Subsequently, they release active mediators, such as histamine, leukotrienes, and prostaglandin-E2 (PGE2) that play a critical underlying role in allergic reactions [7]. AD is further aggravated by the production of vascular endothelial growth factor-a (VEGFPLOS ONE | www.plosone.org 1 November 2014 | Volume 9 | Issue 11 | e113143 Nanoparticles for Immunomodulation in Atopic Dermatitis systemic absorption compared to other TGs. This further improves its clinical applicability and therapeutic compliance [12]. To further broaden therapeutic feasibility and patient compliance, HC was coadministered with hydroxytyrosol (HT), a powerful oxygen free radical scavenger, skin soother, and wound healer. Successful topical/percutaneous delivery of drugs has been limited due to the penetration barriers provided by the SC [15]. Various active and passive penetration-enhancing approaches, including chemical enhancers [16], electroporation [17], microneedles [18], and several vesicular delivery systems such as colloidal carriers [19], liposomes [20], ethosomes [21], solid lipid nanoparticles [22] and nano-emulsions [23] have been investigated to overcome this problem. Besides, polymeric nanoparticles (NPs) are well recognized as an advanced non-invasive technique to facilitate delivery of therapeutics into the skin [24] without detrimental effect on SC [25,26]. The usefulness of polymeric NPs has also been highlighted by Hussain and co-workers in achieving therapeutic dose in the epidermis and dermis and to reduce systemic absorption of TGs and thus minimizing their side eff (...truncated)