Produção e avaliação de nanoemulsão catiônica como possível vetor não-viral para pIRES2-EGFP

Gene therapy is based on the transfer of exogenous genetic material into cells or tissues in order to correct, supplement or silencing a particular gene. To achieve this goal, efficient vehicles, viral or non-viral, should be developed. The aim of this work was to produce and evaluate a nanoemulsion system as a possible carrier for no-viral gene therapy able to load a plasmid model (pIRES2-EGFP). The nanoemulsion was produced by the sonication method, after been choose in a pseudo-ternary phase diagram build with 5 % of Captex 355®, 1.2 % of Tween 80®, 0.8 % of Span 80®, 0.16% of stearylamine and water (to 100 %). Measurements of droplet size, polydispersity index (PI), zeta potential, pH and conductivity, were performed to characterize the system. Results showed droplets smaller than 200 nm (PI < 0.2) and zeta potential > 30 mV. The formulation pH was near to 7.0 and conductivity was that expected to oil in water systems (70 to 90 μS/s) A scale up study, the stability of the system and the best sterilization method were also evaluated. We found that the system may be scaled up considering the time of sonication according to the volume produced, filtration was the best sterilization process and nanoemulsions were stable by 180 days at 4 ºC. Once developed, the complexation efficiency of the plasmid (pDNA) by the system was tested by agarose gel electrophoresis retardation assay.. The complexation efficiency increases when stearylamine was incorporated into aqueous phase (from 46 to 115 ng/μL); regarding a contact period (nanoemulsion / pDNA) of at least 2 hours in an ice bath, for complete lipoplex formation. The nanoemulsion showed low toxicity in MRC-5 cells at the usual transfection concentration, 81.49 % of survival was found. So, it can be concluded that a nanoemulsion in which a plasmid model was loaded was achieved. However, further studies concerning transfectation efficiency should be performed to confirm the system as non-viral gene carrier

Estudo da eficácia do tensoativo sorbitano tween 80 veiculado em nanoemulsão contendo óleo de soja, como inibidor de corrosão

Nowadays, the use of chemicals that satisfactorily meet the needs of different sectors of the chemical industry is linked to the consumption of biodegradable materials. In this context, this work contemplated biotechnological aspects with the objective of developing a more environmentally-friendly corrosion inhibitor. In order to achieve this goal, nanoemulsion-type systems (NE) were obtained by varying the amount of Tween 80 (9 to 85 ppm) a sortitan surfactant named polyoxyethylene (20) monooleate. This NE-system was analyzed using phase diagrams in which the percentage of the oil phase (commercial soybean oil, codenamed as OS) was kept constant. By changing the amount of Tween 80, several polar NE-OS derived systems (O/W-type nanoemulsion) were obtained and characterized through light scattering, conductivity and pH, and further subjected to electrochemical studies. The interfacial behavior of these NE-OS derived systems (codenamed NE-OS1, S2, S3, S4 and S5) as corrosion inhibitors on carbon steel AISI 1020 in saline media (NaCl 3.5%) were evaluated by measurement of Open Circuit Potential (OCP), Polarization Curves (Tafel extrapolation method) and Electrochemical Impedance Spectroscopy (EIS). The analyzed NE-OS1 and NE-OS2 systems were found to be mixed inhibitors with quantitative efficacy (98.6% - 99.7%) for concentrations of Tween 80 ranging between 9 and 85 ppm. According to the EIS technique, maximum corrosion efficiency was observed for some tested NE-OS samples. Additionaly to the electrochemical studies, Analysis of Variance (ANOVA) and Principal Component Analysis (PCA) were used, characterization of the nanoemulsion tested systems and adsorption studies, respectively, which confirmed the results observed in the experimental analyses using diluted NE-OS samples in lower concentrations of Tween 80 (0.5 1.75 ppm)

Obtenção de novos sistemas nanoautoemulsificantes a partir do óleo das sementes de Azadirachta indica A Juss e suas aplicações como inibidores de corrosão

Corrosion is a natural process that causes progressive deterioration of materials, so, reducing the corrosive effects is a major objective of development of scientific studies. In this work, the efficiency of corrosion inhibition on a AISI 1018 carbon steel of the nanoemulsion system containing the oil of the seeds of Azadirachta indica A. Juss (SNEOAI) was evaluated by the techniques of linear polarization resistance (LPR) and weight loss (CPM), a instrumented cell. For that, hydroalcoholic extract of leaves of A. indica (EAI) was solubilized in a nanoemulsion system (SNEOAI) of which O/W system (rich in aqueous phase). This nanoemulsion system (tested in different concentrations) was obtained with oil from the seeds of this plant species (OAI) (oil phase), dodecylammonium chloride (DDAC) (surfactant), butanol (cosurfactant) and water, using 30 % of C/T (cosurfactant/surfactant), 0.5 % of oil phase and 69.5 % of aqueous phase, and characterized by surface tension, rheology and droplet sizes. This systems SNEOAI and SNEOAI-EAI (nanoemulsion containing hydroalcoholic extract - EAI) showed inhibition efficiencies in corrosive environment in saline (1 %), for the method of LPR with significant value of 70.58 % (300 ppm) to SNEOAI, 74.17 % (100 ppm) and 72.51 % (150 ppm) to SNEOAI-EAI. The best efficiencies inhibitions were observed for the method of CPM with 85.41 % for the SNEOAI (300 ppm) and 83.19 % SNEOAI-EAI (500 ppm). The results show that this formulation could be used commercially for use as a corrosion inhibitor, this research contributed to the biotechnological applicability of Azadirachta indica, considering the large use of this plant species rich in limonoids (tetranortriterpenoids), especially azadirachtin

Análise imunoistoquímica após terapia fotodinâmica com cloro-alumínio ftalocianina em tecidos gengivais de humanos

Although photodynamic therapy have been used as a useful tool over the past 30 years in oncology, few clinical trials have been conducted in dentistry. Photodynamic therapy (PDT) uses non - toxic photosensitizers and selective which are administered in target cells followed by local application of visible light, producing reactive oxygen species capable of causing cell death by apoptosis or necrosis, injured the local vasculature, and exert important effects on the im mune system. New generations of photosensitizing agents, such as nanoparticulate phthalocyanines, has shown excellent results in antitumor and antibacterial activity . In this context, the present work constitutes the first clinical protocol of local appli cation of nanoemulsion chloro - aluminum phthalocyanine (AlClFc) followed by irradiation in human gingiva, and analyzed descriptively and comparatively , by means of immunohistochemistry , the expression of RANK , RANKL , OPG and VEGF in a split - mouth model . Eight healthy volunteers with clinical indication for extraction were included in the study . Seven days before the extraction, was injected in the gingiva of participants, 5 μ M of nanoemulsion AlClFc followed by irra diation with diode laser (660nm , 7 J/cm2 ), the contralateral side was used as control. Tissue specimens were removed seven days after the TFD is performed. Tissues sample were divided into two groups (test and con trol groups) for histological and immunohistochemical analysis. Patients were monitored at days, 0, 7, 14 and 30 to assess adverse effects of the therapy. Vascular alterations were seen in gingival samples that received PDT. Areas of edema and vascular con gestion, and intense vascularization were viewed . Additionally, dystrophic calcification in subepithelial region were observed in the test group. The results showed a similar pattern of immunostaining scores of RANK, RANKL and VEGF between the test and co ntrol groups, with no statistically significant difference (p = 0.317, p = 0.777, p = 0 .814, respectively). RANK and RANKL exhibited weak or absent immunostaining in most specimens analyzed. There was n o immunostaining for OPG. VEGF showed moderate to stro ng immunostaining in specimens from the test group. In addition, the clinical study showed that therapy was well tolerated by all patients. Adverse effects were short - time and completely reversible. Taken together, the results presented in this study showe d that PDT mediated by nanoemulsion containing AlClPc is safe for clinical application in gingival tissue and suggests that a strong immunostaining for VEGF after therapy .

Estudo comparativo de diferentes sistemas de injeção de vapor aplicados à remediação de solos contaminados por diesel

Among the potentially polluting economic activities that compromise the quality of soil and groundwater stations are fuel dealers. Leakage of oil derived fuels in underground tanks or activities improperly with these pollutants can contaminate large areas, causing serious environmental and toxicological problems. The number of gas stations grew haphazardly, without any kind of control, thus the environmental impacts generated by these enterprises grew causing pollution of soil and groundwater. Surfactants using various techniques have been proposed to remedy this kind of contamination. This study presents innovation as the application of different systems containing surfactant in the vapor phase and compares their diesel removal efficiencies of soil containing this contaminant. For this, a system that contains seven injection wells the following vaporized solutions: water, surfactant solution, microemulsion and nanoemulsion, The surfactants used were saponified coconut oil (OCS), in aqueous solution and an ethoxylated alcohol UNTL-90: aqueous solution , and nanoemulsion and microemulsion systems. Among the systems investigated, the nanoemulsion showed the highest efficiency, achieving 88% removal of residual phase diesel, the most ecologically and technically feasible by a system with lower content of active matter

Efeitos da terapia fotodinâmica com aluminio – cloro ftalocianina sobre a peroxidação lipídica e produtos antioxidantes em tecidos inflamados animais

Photodynamic therapy (PDT) consists of a non-toxic photosensitizing agent (FS) administration followed by a laser source resulting in a sequence of photochemical and photobiological processes that generate reactive oxygen species (ROS) that damaging cells. The present work evaluated the effects of PDT nanoemulsion-aluminum chloride phthalocyanine (AlClFc) mediated on malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels, which represent indicators involved in oxidative stress and antioxidant defenses. For this purpose, this study used 120 female rats of the Rattus norvegicus species, Wistar race, divided into 5 groups: Healthy (H), with periodontal disease (PD), with periodontal disease and treatment with FS (F), with periodontal disease and treatment with the laser (L); and periodontal disease and treatment with PDT (FL). An experimental model for represent periodontal disease (PD) was induced by ligature (split-mouth). Seven days later the induction of PD, the treatments were instituted according to the groups. In the group treated with PDT was applied 40μl FS (5μM) followed by laser irradiation diode InGaAlP (660nm, 100J / cm2). The rats were sacrificed on the 7th and 28th day after treatment and tissue specimens were removed and subjected to histological, immunohistochemical methods and enzymatic colorimetric measurements with detection by UV / VIS spectroscopy. Inflammatory changes, connective tissue disorganization and alveolar bone loss were displaying in groups with PD induced. The enzyme dosages showed that MDA levels were higher in PD induced groups, with no statistically significant differences (p> 0.05). High levels of GSH were found in groups L (p = 0.028) and FL (p = 0.028) compared with PD group, with statistically significant differences. Immunohistochemistry for SOD showed higher immunostaining in L and FL groups, compared to the PD group without statistically significant differences (p> 0.05). GPx showed lower immunoreactivity in the DP group when compared to the other groups and statistically significant differences were observed between the DPxL groups (p <0.05). TFD administered in this experiment did not induce elevation of MDA levels significantly increased the GSH levels and showed intense immunostaining pada SOD and GPx, showing that this therapy does not accentuated lipid peroxidation, however, it was able to induce effects on the antioxidant defenses processes. The LBI therapy appeared to show photomodulatory promoting effects reduction of the MDA levels, increasing GSH levels and with intense immunostaining for SOD and GPx, demonstrating that laser therapy induced antioxidant effects.