Effects of andiroba oil (Carapa guianensis aublet) on the immune system in inflammation and wound healing: A scoping review.

ETHNOPHARMACOLOGICAL RELEVANCE: Andiroba seed oil (Carapa guianensis Aubl.) is widely used by traditional populations in tropical countries, especially in the Brazilian Amazon, because of its anti-inflammatory, antirheumatic, antiseptic, healing and antipyretic properties, among others, which makes it useful for the treatment, mainly, of skin afflictions and wounds. AIM OF THE STUDY: To describe the modulation of the immune system by andiroba oil (Carapa guianensis Aubl.) in inflammation and wound healing. MATERIALS AND METHODS: A scoping review was performed, following the recommendations of the Joanna Briggs Institute (JBI) and PRISMA for Scoping Reviews (PRISMA-ScR). As inclusion criteria, in vitro, in vivo, ex vivo, and clinical studies were selected, in Portuguese, English, or Spanish, in thirteen databases of published studies, gray literature, and references of the included studies, which deal with immune modulation by andiroba oil in the context of the various therapeutic applications that make use of its anti-inflammatory and wound healing properties. The selection of information sources was carried out by two independent reviewers between November 2022 and January 2023. The process of data extraction and evidence analysis was conducted by four pairs of independent reviewers between January and February 2023. RESULTS: 22 sources of evidence were included in this scoping review, mostly scientific articles published between 2005 and 2021 with in vivo sampling. The evidence suggests that andiroba oil reduces inflammation and promotes the healing of wounds of multiple etiologies by reducing leukocyte infiltration, increasing phagocytic activity, enhancing interleukin and inflammatory cytokine activity, promoting fibroblast recovery, increasing growth factors, reducing apoptotic cells, promoting reepithelialization, as well as promoting angiogenesis, reducing edema, and stimulating the production of glucocorticoids that alleviate pain. Additionally, different formulations of the oil (such as nanoemulsions, films and gels) are more effective in modulating inflammation and wound healing compared to in natura oil. CONCLUSIONS: Evidence in the literature suggests that andiroba oil (Carapa guianensis Aubl.) has positive effects on immune modulation in inflammation and wound healing, which makes it a biocompound with high therapeutic potential.

Synthesis of copaiba (Copaifera officinalis) oil nanoemulsion and the potential against Zika virus: An in vitro study.

Zika virus (ZIKV) has spread all over the world since its major outbreak in 2015. This infection has been recognized as a major global health issue due to the neurological complications related to ZIKV infection, such as Guillain-Barré Syndrome and Zika virus Congenital Syndrome. Currently, there are no vaccines or specific treatments for ZIKV infection, which makes the development of specific therapies for its treatment very important. Several studies have been developed to analyze the potential of compounds against ZIKV, with the aim of finding new promising treatments. Herein, we evaluate the ability of a copaiba (Copaifera officinalis) oil nanoemulsion (CNE) to inhibit ZIKV. First, the highest non-cytotoxic concentration of 180 µg/mL was chosen since this concentration maintains 80% cell viability up to 96h after treatment with CNE in VERO cells resulted from MTT assay. The intracellular uptake assay was performed, and confirmed the internalization of the nanoemulsion in cells at all times analyzed. VERO cells were infected with ZIKV and simultaneously treated with CNE and the nanoformulation without oil (ENE) at the highest non-toxic concentration. The results evaluated by plaque assay revealed a viral inhibition of 80% for CNE and 70% for ENE. A dose-dependence assay revealed that the CNE treatment demonstrated a dose-dependent response in the viral RNA levels, whereas all ENE tested concentrations exhibited a similar degree of reduction. Taken together, our results suggest CNE as a promising nano-sized platform to be further studied for antiviral treatments.

A novel N95 respirator with chitosan nanoparticles: mechanical, antiviral, microbiological and cytotoxicity evaluations.

BACKGROUND: It is known that some sectors of hospitals have high bacteria and virus loads that can remain as aerosols in the air and represent a significant health threat for patients and mainly professionals that work in the place daily. Therefore, the need for a respirator able to improve the filtration barrier of N95 masks and even inactivating airborne virus and bacteria becomes apparent. Such a fact motivated the creation of a new N95 respirator which employs chitosan nanoparticles on its intermediate layer (SN95 + CNP). RESULTS: The average chitosan nanoparticle size obtained was 165.20 ± 35.00 nm, with a polydispersity index of 0.36 ± 0.03 and a zeta potential of 47.50 ± 1.70 mV. Mechanical tests demonstrate that the SN95 + CNP respirator is more resistant and meets the safety requisites of aerosol penetration, resistance to breath and flammability, presenting higher potential to filtrate microbial and viral particles when compared to conventional SN95 respirators. Furthermore, biological in vitro tests on bacteria, fungi and mammalian cell lines (HaCat, Vero E6 and CCL-81) corroborate the hypothesis that our SN95 + CNP respirator presents strong antimicrobial activity and is safe for human use. There was a reduction of 96.83% of the alphacoronavirus virus and 99% of H1N1 virus and MHV-3 betacoronavirus after 120 min of contact compared to the conventional respirator (SN95), demonstrating that SN95 + CNP have a relevant potential as personal protection equipment. CONCLUSIONS: Due to chitosan nanotechnology, our novel N95 respirator presents improved mechanical, antimicrobial and antiviral characteristics.

Combination of selol nanocapsules and magnetic hyperthermia hinders breast tumor growth in aged mice after a short-time treatment.

Short time treatment with reduced dosages of selol-loaded PLGA nanocapsules (NcSel) combined with magnetic hyperthermia (MHT) is evaluated in aged Erhlich tumor-bearing mice. Clinical, hematological, biochemical, genotoxic and histopathological parameters are assessed during 7 d treatment with NcSel and MHT, separately or combined. The time evolution of the tumor volume is successfully modeled using the logistic mathematical model. The combined therapy comprising NcSel and MHT is able to hinder primary tumor growth and a case of complete tumor remission is recorded. Moreover, no metastasis was diagnosed and the adverse effects are negligible. NcSel plus MHT may represent an effective and safe alternative to cancer control in aged patients. Future clinical trials are encouraged.

Oily core/amphiphilic polymer shell nanocapsules change the intracellular fate of doxorubicin in breast cancer cells.

The aim of this work was to develop and test the in vitro biological activity of nanocapsules loaded with a doxorubicin (DOX) free base dissolved in a core of castor oil shelled by poly(methyl vinyl ether-co-maleic anhydride) conjugated to n-octadecylamine residues. This system was stable and monodisperse, with a hydrodynamic diameter of about 300 nm. These nanocapsules changed the intracellular distribution of DOX, from the nuclei to the cytoplasm, and exhibited higher toxicity towards cancer cells - 4T1 and MCF-7 - and significantly lower toxicity towards normal cells - NIH-3T3 and MCF-10A - in vitro. In conclusion, these nanocapsules are suitable DOX carriers, which remain to be studied in in vivo tumor models.

Optimizing liposomes for delivery of Bowman-Birk protease inhibitors - Platforms for multiple biomedical applications.

One of the major challenges in the administration of therapeutic proteins involves delivery limitations. Liposomes are well-known drug delivery systems (DDS) that have been used to overcome this drawback; nevertheless, low protein entrapment efficiency (EE) still limits their wide biomedical application on a commercial scale. In the present work, different methods for protein entrapment into liposomes were tested in order to obtain tailored DDS platforms for multiple biomedical applications. The protein used as model was the Black-eyed pea Trypsin and Chymotrypsin Inhibitor (BTCI), a member of the Bowman-Birk protease inhibitor family (BBIs), which has been largely explored for its potential application in many biomedical therapies. We optimized reverse-phase evaporation (REV) and freeze/thaw (F/T) entrapment methods, using a cationic lipid matrix to entrap expressive amounts of BTCI (∼100 µM) in stable liposomes without affecting its protease inhibition activity. The influence of various parameters (e.g. entrapment method, liposome composition, buffer type) on particle size, charge, polydispersity, and EE of liposomes was investigated to provide an insight on how to control such parameters in view of obtaining a high entrapment yield. In addition, BTCI liposome platforms obtained herein showed to be versatile vesicles, allowing surface modification with moieties/polymers of interest (e.g. PEG, transferrin). The aforementioned results are relevant to focusing on the entrapment of other promising BBIs or protein agents sharing similar structural features. These findings encourage future studies to investigate the advantages of using the liposome platforms presented herein to broaden the use of this type of DDS for BBI biomedical applications.

Simple and Selective HPLC-UV/Vis Bioanalytical Method to Determine Aluminum Phthalocyanine Chloride in Skin Permeation Studies.

Considering the feasibility of the aluminum phthalocyanine chloride (AlPcCl) application in the topical photodynamic therapy of cutaneous tumors and the lack of HPLC methods capable of supporting skin permeation experiments using this compound, the aim of this study was to obtain a simple and selective chromatographic method for AlPcCl determination in skin matrices. A HPLC-UV/Vis method was developed using a normal-phase column operating at 30°C, an isocratic mobile phase of methanol : phosphoric acid (0.01 M) at 1.5 mL/min, and detection at 670 nm. The method exhibited (i) selectivity against various contaminants found in the different skin layers, (ii) high drug extraction capacity from the hair follicle (>70%) and remaining skin (>80%), and (iii) low limits of detection and of quantification (0.03 and 0.09 µg/mL, resp.). The method was also linear in the range from 0.1 to 5.0 µg/mL (r = 0.9994) and demonstrated robustness with regard to experimental chromatographic parameters according to a factorial design. Lastly, the developed method was successfully tested in in vitro skin permeation studies of AlPcCl, proving its effectiveness in the development of pharmaceutical delivery systems containing this drug for topical photodynamic therapy of skin cancers.

Nanocapsules for the co-delivery of selol and doxorubicin to breast adenocarcinoma 4T1 cells in vitro.

Nanocapsules (NCS-DOX) with an oily core of selol and a shell of poly(methyl vinyl ether-co-maleic anhydride) covalently conjugated to doxorubicin were developed. These nanocapsules are spherical, with an average hydrodynamic diameter of about 170 nm, and with negative zeta potential. NCS-DOX effectively co-delivered the selol and the doxorubicin into 4T1 cells and changed the intracellular distribution of DOX from the nuclei to the mitochondria. Moreover, a significantly increased cytotoxicity against 4T1 cells was observed, which is suggestive of additive or synergic effect of selol and doxorubicin. In conclusion, PVM/MA nanocapsules are suitable platforms to co-deliver drugs into cancer cells.

Nanostructured Systems for the Organelle-specific Delivery of Anticancer Drugs.

Nanotechnology has provided powerful tools to improve the chemotherapy of cancer. Different nanostructures have been developed which deliver the anticancer drugs more selectively to tumor than to healthy tissues. The result has generally been the increase in efficacy and safety of classical anticancer drugs. In recent years, several studies have focused not only on the delivery of anticancer drugs to tumors, but also on delivering the drugs to specific organelles of cancer cells. Endoplasmic reticulum, Golgi apparatus, lysosomes, mitochondria, and nucleus have been the targets of different nanostructured drug delivery systems developed with the goal of circumventing drugresistance, increasing drug efficacy, and so on. So far, the results described in the literature show that this strategy may be used to improve chemotherapy outcomes. In this review a discussion is presented on the strategies described in the literature to deliver anticancer drugs to specific organelles of cancer cells by using nanostructures.

Cytotoxic Activity and Antiproliferative Effects of Crude Skin Secretion from Physalaemus nattereri (Anura: Leptodactylidae) on in vitro Melanoma Cells.

Anuran secretions are rich sources of bioactive molecules, including antimicrobial and antitumoral compounds. The aims of this study were to investigate the therapeutic potential of Physalaemus nattereri skin secretion against skin cancer cells, and to assess its cytotoxic action mechanisms on the murine melanoma cell line B16F10. Our results demonstrated that the crude secretion reduced the viability of B16F10 cells, causing changes in cell morphology (e.g., round shape and structure shrinkage), reduction in mitochondrial membrane potential, increase in phosphatidylserine exposure, and cell cycle arrest in S-phase. Together, these changes suggest that tumor cells die by apoptosis. This skin secretion was also subjected to chromatographic fractioning using RP-HPLC, and eluted fractions were assayed for antiproliferative and antibacterial activities. Three active fractions showed molecular mass components in a range compatible with peptides. Although the specific mechanisms causing the reduced cell viability and cytotoxicity after the treatment with crude secretion are still unknown, it may be considered that molecules, such as the peptides found in the secretion, are effective against B16F10 tumor cells. Considering the growing need for new anticancer drugs, data presented in this study strongly reinforce the validity of P. nattereri crude secretion as a rich source of new anticancer molecules.

Physico-chemical characterization and cytotoxicity evaluation of curcumin loaded in chitosan/chondroitin sulfate nanoparticles.

In this study, chitosan (CTS)/chondroitin sulfate (CS) nanoparticles, both pure and curcumin-loaded, were synthesized by ionic gelation. This method is simple and efficient for obtaining nanoparticles with a low polydispersity index (0.151±0.03 to 0.563±0.07) and hydrodynamic diameter in the range of 175.7±2.5 to 710.2±8.9nm, for this study. Samples have a relatively high zeta potential value, a fact that indicates that the colloidal system has good physical and chemical stabilities. The efficiency of the curcumin encapsulation in nanoparticles, which ranged from 62.4±0.61% to 68.3±0.88%, depends on the pH of the chitosan solution. The release of curcumin from the nanoparticles was enabled by a diffusion mechanism, with fast release in a phosphate buffer solution at pH6.8. The assaying of cell viability by the MTT test showed that the presence of both free curcumin and curcumin in the nanoencapsulated form leads to a statistically significant reduction in the viability of A549 cells, by comparison with the control group. The most significant reductions in cell viability of 41.1% and 60.4% (p<0.0001) were observed after 72h, by using 40µmol∙L(-1) free curcumin and curcumin encapsulated in CTS/CS nanoparticles with the chitosan solution at pH6.0, respectively.

In vitro cytotoxic activity of chitosan-bullfrog oil microemulsion against melanoma cells.

Microemulsion-based animal oils, alone or associated with polymers have been extensively used in pharmacy, medicine and cosmetics, since the major lipid constituents of the oils show several biological activities. Despite showing antimicrobial activity, there are no reports in the literature regarding the effects of bullfrog oil on cytotoxic activity against tumor cells. The aim of the present study was to synthesize, characterise and evaluate the in vitro effects on melanoma cell line (B16F10) of bullfrog oil microemulsions associated or not with chitosan, surfactant and bullfrog oil (CSBO) and surfactant and bullfrog oil (SBO), respectively. The microemulsions were developed and their physical-chemical characteristics were evaluated by light microscopy, dynamic light scattering, atomic force microscopy and zeta potential. The microemulsions showed regular spherical shapes, high polydispersity and excellent (+82.2 ± 1.0 mV) to low (-16.0 ± 0.5 mV), colloidal stability. The systems significantly decreased the in vitro cell viability of melanoma skin cancer by up to 90.2% (CSBO) and 91.8% (SBO); while free bullfrog oil showed no effects. The results obtained from microemulsions of bullfrog oil indicate the potential of the microemulsions developed, alone or in combination with other chemotherapeutic agents, for future use in biomedical approaches aiming towards cancer therapy.

Antibacterial, antibiofilm and cytotoxic activities of Terminalia fagifolia Mart. extract and fractions.

BACKGROUND: The methicillin resistance of bacteria from the genus Staphylococcus and its ability to form biofilms are important factors in pathogenesis of these microorganisms. Thus, the search for new antimicrobials agents, especially from plants, has been intensified. In this context, Terminalia species have been the subject of research for many pharmacological activities. In this study we evaluated the antibacterial, antibiofilm and cytotoxic activities of the ethanol extract (EtE) from Terminalia fagifolia stem bark as well as that of three fractions of the extract (AqF, HaF and WSF). METHODS: We determined the minimum inhibitory concentration (MIC) by microdilution in 96-well plates, where the strains were exposed to serial dilutions of the ethanol extract and fractions, ranging from 12.5 to 400 µg/mL. We then determined the minimum bactericidal concentration (MBC), seeding the inoculum (10 µL) with concentrations equal to or greater than the MIC in Mueller-Hinton agar. To test the antibiofilm activity biofilm formation was induced in the presence of concentrations equivalent to 1/2, 1/4 and 1/8 of the MIC extract or fraction tested. In addition, the effect of the EtE and the fractions on cell viability was tested by the MTT assay on human MCF-7 breast cancer and mouse fibroblast NIH/3T3. To obtain high-resolution images of the effect of the aqueous fraction on the bacterial morphology, atomic force microscopy (AFM) imaging of treated S. aureus cells was performed. RESULTS: We observed antibacterial activity of EtE and fractions with MICs ranging from 25-200 µg/mL and MBCs ranging from 200-400 µg/mL. Regarding antibiofilm activity, both the EtE as the AqF, HaF and WSF fractions showed significant inhibition of the biofilm formation, with inhibition of biofilms formation of over 80% for some strains. The EtE and fractions showed a moderate cytotoxicity in cell line NIH/3T3 viability and potential antitumoral activity on human breast cancer cell line MCF-7. The microscopic images obtained revealed morphological changes to the S. aureus ATCC 29213 surface caused by AqF, as well as significant size alterations. CONCLUSIONS: The results show potential antibacterial, antibiofilm and antitumoral activities of the ethanol extract and fractions of T. fagifolia.

PVM/MA-shelled selol nanocapsules promote cell cycle arrest in A549 lung adenocarcinoma cells.

BACKGROUND: Selol is an oily mixture of selenitetriacylglycerides that was obtained as a semi-synthetic compound containing selenite. Selol is effective against cancerous cells and less toxic to normal cells compared with inorganic forms of selenite. However, Selol's hydrophobicity hinders its administration in vivo. Therefore, the present study aimed to produce a formulation of Selol nanocapsules (SPN) and to test its effectiveness against pulmonary adenocarcinoma cells (A549). RESULTS: Nanocapsules were produced through an interfacial nanoprecipitation method. The polymer shell was composed of poly(methyl vinyl ether-co-maleic anhydride) (PVM/MA) copolymer. The obtained nanocapsules were monodisperse and stable. Both free Selol (S) and SPN reduced the viability of A549 cells, whereas S induced a greater reduction in non-tumor cell viability than SPN. The suppressor effect of SPN was primarily associated to the G2/M arrest of the cell cycle, as was corroborated by the down-regulations of the CCNB1 and CDC25C genes. Apoptosis and necrosis were induced by Selol in a discrete percentage of A549 cells. SPN also increased the production of reactive oxygen species, leading to oxidative cellular damage and to the overexpression of the GPX1, CYP1A1, BAX and BCL2 genes. CONCLUSIONS: This study presents a stable formulation of PVM/MA-shelled Selol nanocapsules and provides the first demonstration that Selol promotes G2/M arrest in cancerous cells.

Apoptosis and lysosome membrane permeabilization induction on breast cancer cells by an anticarcinogenic Bowman-Birk protease inhibitor from Vigna unguiculata seeds.

In this work, we report the effects of a Bowman-Birk protease inhibitor, the Black-Eyed Pea Trypsin/Chymotrypsin Inhibitor - BTCI, purified from Vigna unguiculata seeds, on the MCF-7 breast cancer cells. The treatment of MCF-7 with 200microM BTCI for 72h induced significant reduction of the cell viability and proliferation (arrest in S and G2/M phase). These cytostatic effects were accompanied by acute morphological modifications including the alteration of the nuclear morphology, plasma membrane fragmentation, cytoplasm disorganization, presence of double-membrane vesicles, mitochondrial swelling, and an increase in the size of lysosomes. Significative DNA fragmentation, annexin-V(+) cell number increase, mitochondrial membrane potential reduction, and cytoplasm acidification were also detected. All together, these cytostatic and cytotoxic results point out to BTCI-induced apoptosis cell death associated with severe cell morphological alterations and lysosome membrane permeabilization. Our study confirms the anticarcinogenic potential of Bowman-Birk protease inhibitors and identifies BTCI as a promising tool for drug developments aimed at the treatment of breast cancer.