Abstract
Annatto (Bixa orellana L.) is extensively used as food pigment worldwide. Recently, several studies have found it to have healing and antioxidant properties, as well as effective action against leishmaniasis. Therefore, the purpose of this study was to incorporate the oil obtained from annatto seeds into a nanostructured lipid carrier (NLC) and evaluate its physicochemical properties and biological activity against Leishmania major. Nanoparticles were prepared by the fusion-emulsification and ultrasonication method, with the components Synperonic™ PE (PL) as the surfactant, cetyl palmitate (CP) or myristyl myristate (MM) as solid lipids, annatto oil (AO) (2% and 4%, w/w) as liquid lipid and active ingredient, and ultra-pure water. Physicochemical and biological characterizations were carried out to describe the NLCs, including particle size, polydispersity index (PDI), and zeta potential (ZP) by dynamic light scattering (DLS), encapsulation efficiency (EE%), thermal behavior, X-ray diffraction (XRD), transmission electron microscopy (TEM), Electron Paramagnetic Resonance (EPR), cytotoxicity on BALB/c 3T3 fibroblasts and immortalized human keratinocyte cells, and anti-leishmaniasis activity in vitro. Nanoparticles presented an average diameter of ~200 nm (confirmed by TEM results), a PDI of less than 0.30, ZP between −12.6 and −31.2 mV, and more than 50% of AO encapsulated in NLCs. Thermal analyses demonstrated that the systems were stable at high temperatures with a decrease in crystalline structure due to the presence of AOs (confirmed by XRD). In vitro, the anti-leishmania test displayed good activity in encapsulating AO against L. major. The results indicate that the oily fraction of Bixa orellana L. in NLC systems should be evaluated as a potential therapeutic agent against leishmaniasis.
Highlights
More than one billion people in the world have been affected by NeglectedTropical Diseases (NTDs), which is considered to be an important public health issue [1,2].Leishmaniasis is an emerging uncontrolled NeglectedTropical Diseases (NTDs) [3]
There are different kinds of Leishmania infection: Visceral Leishmaniasis (VL) and American Cutaneous Leishmaniasis (ACL), which is subdivided into Cutaneous Leishmaniasis (CL), Diffuse Cutaneous Leishmaniasis (DCL), and Mucosal Leishmaniasis (ML) [6]
The preparation of lipid nanoparticles involved five steps as presented in Figure 1: (1) Heating: cetyl palmitate (CP) and myristyl myristate (MM) were heated at 10 ◦ C above their melting point, and for nanostructured lipid carrier (NLC), annatto oil fraction (AO) was solubilized into the lipid previously melted (CM or MM)
Summary
More than one billion people in the world have been affected by NeglectedTropical Diseases (NTDs), which is considered to be an important public health issue [1,2].Leishmaniasis is an emerging uncontrolled NTD [3]. More than one billion people in the world have been affected by Neglected. Tropical Diseases (NTDs), which is considered to be an important public health issue [1,2]. Leishmaniasis is an emerging uncontrolled NTD [3]. There are twelve million people who are currently infected worldwide, culminating in a high mortality rate Epidemiological research indicates that there are 350 million people. Pharmaceutics 2021, 13, 1912 at risk from a zoonotic infection disease caused by Leishmania parasites in 102 countries, areas, or territories, and large parts of the world, including the European continent [2,4]. In. Spain, for example, leishmaniasis is endemic and of increasing incidence [5]. There are different kinds of Leishmania infection: Visceral Leishmaniasis (VL) and American Cutaneous Leishmaniasis (ACL), which is subdivided into Cutaneous Leishmaniasis (CL), Diffuse Cutaneous Leishmaniasis (DCL), and Mucosal Leishmaniasis (ML) [6]
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