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Myracrodruon urundeuva (Engl.) Fr. All., commonly known as "aroeira-do-sertão", is a medicinal plant from Anacardiaceae family. In this study, the chemical composition of M. urundeuva essential oil (MuEO) was evaluated by gas chromatography-mass spectrometry (GC-MS), as well as its anti-Leishmania potential, cytotoxicity, and macrophage activation capability as possible antiprotozoal mechanism of action were assessed. Fourteen compounds were identified, which constituted 94.87% of total oil composition. The most abundant components were monoterpenes (80.35%), with β-myrcene (42.46%), α-myrcene (37.23%), and caryophyllene (4.28%) as the major constituents. The MuEO inhibited the growth of promastigotes (IC 205 ± 13.4 μg mL), axenic amastigotes (IC 104.5 ± 11.82 μg mL) and decreased percentage of macrophage infection and number of amastigotes per macrophage (IC of 44.5 ± 4.37 μg⋅mL), suggesting significant anti-Leishmania activity. The cytotoxicity of MuEO was assessed by MTT test in Balb/c murine macrophages and by human erythrocytes lysis assay and low cytotoxicity for these cells was observed. The CC value against macrophages were 550 ± 29.21 μg mL, while cytotoxicity for erythrocytes was around 20% at the highest concentration assessed, with HC > 800 μg mL. While MuEO-induced anti-Leishmania activity is not mediated by increases in both lysosomal activity and nitric oxide production in macrophages, the results suggest the antiamastigote activity is associated with an immunomodulatory activity of macrophages due to an increase of phagocytic capability induced by MuEO. Thus, MuEO presented significant activity against Leishmania amazonensis, probably modulating the activation of macrophages, with low cytotoxicity to murine macrophages and human erythrocytes.

BackgroundTissue engineering has been shown to exhibit great potential for the creation of biomaterials capable of developing into functional tissues. Cellular expansion and integration depends on the quality and surface-determinant factors of the scaffold, which are required for successful biological implants. The objective of this research was to characterize and evaluate the in vitro characteristics of rabbit bone marrow mesenchymal stem cells (BM-MSCs) associated with a bacterial cellulose membrane (BCM). We assessed the adhesion, expansion, and integration of the biomaterial as well as its ability to induce macrophage activation. Finally, we evaluated the cytotoxicity and toxicity of the BCM.MethodsSamples of rabbit bone marrow were collected. Mesenchymal stem cells were isolated from medullary aspirates to establish fibroblast colony-forming unit assay. Osteogenic, chondrogenic, and adipogenic differentiation was performed. Integration with the BCM was assessed by scanning electron microscopy at 1, 7, and 14 days. Cytotoxicity was assessed via the production of nitric oxide, and BCM toxicity was assessed with the MTT assay; phagocytic activity was also determined.ResultsThe fibroblastoid colony-forming unit (CFU-F) assay showed cells with a fibroblastoid morphology organized into colonies, and distributed across the culture area surface. In the growth curve, two distinct phases, lag and log phase, were observed at 15 days. Multipotentiality of the cells was evident after induction of osteogenic, chondrogenic, and adipogenic lineages. Regarding the BM-MSCs’ bioelectrical integration with the BCM, BM-MSCs were anchored in the BCM in the first 24 h. On day 7 of culture, the cytoplasm was scattered, and on day 14, the cells were fully integrated with the biomaterial. We also observed significant macrophage activation; analysis of the MTT assay and the concentration of nitric oxide revealed no cytotoxicity of the biomaterial.ConclusionThe BCM allowed the expansion and biointegration of bone marrow progenitor cells with a stable cytotoxic profile, thus presenting itself as a biomaterial with potential for tissue engineering.

Leishmaniasis is an infectious disease complex caused by protozoa from the Leishmania genus, which presents a broad spectrum of clinical manifestations: cutaneous, mucocutaneous and visceral forms. The current treatments are unsatisfactory considering that few drugs are available and present some level of toxicity. Many lignans and neolignans have been used for the development of new antileishmania drugs. The capability in vitro of the neolignan 2,3-dihydrobenzofuran (2,3-DBF), a commonly found constituent of propolis and other plants, to inhibit the growth of promastigote and macrophage-internalized amastigote forms of Leishmania amazonensis was investigated. The cytotoxicity of this compound was assessed by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) test in BALB/c murine macrophages and human erythrocyte lysis assay. The 2,3-DBF was active against promastigote (IC 50 =1.042 lM) and amastigote (IC 50 =1.43 lM) forms, indicating a potent antileishmanial effect. There was no evidence of cytotoxicity to macrophages or erythrocytes at concentrations ranging from 13 to 0.5 lM, after 48 hr of exposure. The antileishmanial activity is probably mediated by the activation of macrophages, because treatment with 2,3-DBF increases both phagocytic and lysosomal activities, as well as the nitrite (NO 2 À ) levels. These results suggest that 2,3-DBF may be a potential candidate for the development of a new promising antileishmanial drug. Further studies are needed to determine its potential in vivo effect as well as additional mechanisms underlying the antileishmanial and immunomodulatory activities.

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The objective of this study was to evaluate the agronomic characteristics of Nopalea cochenillifera (L.) Salm-Dick under different frequencies and levels of nitrogen application (N). Three nitrogen fertilizer application frequencies (1 time year-1, 6 times year-1 and 12 times year-1) and four nitrogen levels (0, 100, 200 and 400 kg of N ha-1 year-1) were evaluated one year after the planting, in a randomized blocks design with 12 replications. There was no interaction (P>0.05) between application frequencies and nitrogen levels. The number of cladodes, and the green and dry forage mass yields of the cactus pear were higher with the application frequency of 6 times year-1, presenting 18.4 ± 2.0 cladodes per plant, 151.0 ± 17.8 and 9.8 ± 1.0 t ha-1, respectively. The yield doubled with the application of 400 kg N ha-1, when compared to the treatment with no nitrogen application. With the nitrogen application frequency of 1 time year-1 the crude protein and neutral detergent fiber contents were higher, presenting 4.0 ± 0.1% and 33.8 ± 0.5%, respectively. When nitrogen is applied 6 times year-1 it provides higher growth and yield, but lower protein and fiber contents. The level 400 kg N ha-1 provides higher yield and lower nitrogen use efficiency.

The development of ‘smart’ scaffolds has achieved notoriety among current prospects for bone repair, especially for chronic osteopathy, such as osteoporosis. Millions of individuals in the world suffer from poor bone healing due to osteoporosis. The objective of this work was to produce and characterize castor polyurethane (PU) scaffolds (Ricinus communis L.) and evaluate its in vitro biocompatibility with stem cells and osteoinductive effect in vivo on bone failures in a leporid model of osteoporosis. The material was characterized using Fourier-transform infrared spectroscopy, thermogravimetric analysis, SEM, and porosity analysis. Then, the biocompatibility was assessed by adhesion using SEM and cytotoxicity in a 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium assay. The osteoinductive effect in vivo was determined in bone defects in rabbit tibias (Oryctolagus cuniculus) submitted to castor PU scaffold, castor PU scaffold associated with stem cells, and negative control, after four and eight weeks, evaluated by computed microtomography and histopathology. The scaffolds were porous, with an average pore size of 209.5 ± 98.2 µm, absence of cytotoxicity, and positive cell adhesiveness in vitro. All the animals presented osteoporosis, characterized by multifocal osteoblastic inactivity and areas of mild fibrosis. There were no statistical differences between these treatments in the fourth week of treatment. In the eighth week, the treatment with castor PU scaffold alone induced more significant bone formation when compared to the other groups, followed by treatment with an association between castor PU scaffold and stem cells. The castor PU scaffold was harmless to cell culture, favoring cell adhesiveness and proliferation, in addition to inducing bone neoformation in osteoporotic rabbits.

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