Original Article
Assessment of β-lapachone loaded in lecithin-chitosan nanoparticles for the topical treatment of cutaneous leishmaniasis in L. major infected BALB/c mice

https://doi.org/10.1016/j.nano.2015.07.011Get rights and content

Abstract

Patients affected by cutaneous leishmaniasis need a topical treatment which cures lesions without leaving scars. Lesions are produced not only by the parasite but also by an uncontrolled and persistent inflammatory immune response. In this study, we proposed the loading of β-lapachone (β-LP) in lecithin-chitosan nanoparticles (NP) for targeting the drug to the dermis, where infected macrophages reside, and promote wound healing. Although the loading of β-LP in NP did not influence the drug antileishmanial activity it was critical to achieve important drug accumulation in the dermis and permeation through the skin. When topically applied in Leishmania major infected BALB/c mice, β-LP NP achieved no parasite reduction but they stopped the lesion progression. Immuno-histopathological assays in CL lesions and quantitative mRNA studies in draining lymph nodes confirmed that β-LP exhibited anti-inflammatory activity leading to the down-regulation of IL-1β and COX-2 expression and a decrease of neutrophils infiltrate.

From the Clinical Editor

Cutaneous leishmaniasis often leaves patients with unsightly scars due to the body's inflammatory response to the infection. The authors in this paper described topical treatment using β-lapachone (β- LP) loaded in lecithin-chitosan nanoparticles (NP) in an animal model. Results confirmed the reduction of inflammatory response without affecting the parasite killing efficacy. These findings would pave way for further clinical testing in the near future.

Graphical abstract

After demonstrating the ex vivo performance of lecithin-chitosan NP for enhancing the penetration and dermal accumulation of the poorly soluble drug β-lapachone, the efficacy of the treatment was topically assayed in L. major infected BALB/c mice. Despite the treatment did not influence the parasite burden, it down-regulated IL-1β and COX-2 and reduced the neutrophil infiltrate, thus decreasing the necrosis and size of the skin lesion. Our finding offers a new approach of utility for weakening skin damage in cutaneous leishmaniasis lesions.

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Section snippets

Preparation and characterization of β-lapachone nanoparticles

β-LP NP were prepared following the procedure reported by Sonvico et al. with modifications.18 Briefly, chitosan (95/100, 1% w/v) was solubilized in 2% w/v lactic acid whereas β-LP (10 mg/mL) was dissolved in an ethanolic solution of LEC. The organic phase was prepared by mixing LEC:DDAB solutions (60:30 mM, respectively) or LEC:MNL solutions (75:45 mM, respectively) in ethanol:acetone (5:4 v/v). Afterwards, β-LP NP suspensions were obtained by injection of 4 mL of this organic mixture into 4 mL of

Preparation and characterization of β-lapachone nanoparticles

LEC-chitosan NP containing MNL or DDAB were prepared and, particle size (Ø), polydispersity index (PDI), Z-potential and encapsulation efficiency (EE) were determined in order to characterize the NP (Table 1). The mean size of the NP prepared for this study ranged from 317 to 574 nm, showing LEC:DDAB NP smaller sizes than LEC:MNL NP. The Z-potential value was higher for the NP containing DDAB (around 90 mV) than those containing MNL (around 30 mV) but in all cases they were positively charged.

Discussion

Looking for a better CL topical therapy and aiming to explore the goodness of NP skin delivery in CL lesions, in the current study, β-LP was encapsulated in NP containing LEC, a safe and biocompatible excipient widely used in delivery systems, as well as chitosan, an antimicrobial agent with penetration-enhancing properties. Moreover, DDAB and MNL, two different lipids with antimicrobial activity20, 21 and different charge, were also included into the formulations to investigate if they could

Acknowledgements

We would like to thank Laura Guembe (Servicio de Morfología, Centro para la Investigación Médica Aplicada, CIMA), Ainhoa Urbiola and Cristina Ederra (Unidad de Imagen, CIMA) for their excellent technical support with imaging techniques.

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    Conflict of interest: The authors declare no conflict of interest.

    Funding information: This study has been supported by the FIMA Foundation (Fundación para la Investigación Médica Aplicada) and the Institute of Tropical Health. J.S. received a grant of ADA Foundation (Asociación de Amigos, University of Navarra) and E.M. was awarded by the Ministry of Economy and Competitiveness of the Spanish Government (Subprograma: Torres Quevedo).

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    The authors have contributed equally to this work.

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