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Item Alternativas tecnológicas para aumentar a biodisponibilidade de anti-inflamatórios: gel mucoadesivo termorreversível e aplicação de sílica mesoporosa para amorfização(Universidade Federal de Goiás, 2020-03-20) Antonino, Rayane Santa Cruz Martins de Queiroz; Nascimento, Thais Leite; http://lattes.cnpq.br/4065607913504134; Lima, Eliana Martins; http://lattes.cnpq.br/7248774319455970; Lima, Eliana Martins; Alonso, Christian Gonçalves; Gil, Eric de Souza; Silva, Luís Antônio Dantas; Brito, Wesley de AlmeidaImproving the therapeutic response of drugs is one of the major goals of pharmaceutical technology. This large area of research and development uses multidisciplinary technologies and knowledge aimed at optimizing drug delivery systems. It seeks, for example, to reduce side effects with the application of nanoparticulate systems (Chung et al., 2019), to improve the reach of drugs in the central nervous system (CNS) through new delivery and administration systems (Qureshi et al., 2019), increase the delivery and therapeutic action of drugs by promoting longer drug contact time at the site of action, such as the development of mucoadhesive formulations for the treatment of inflamed mucous membranes (Léber et al., 2019). It also seeks to increase the bioavailability of drugs with low aqueous solubility by obtaining the amorphous form of the drug either through porous adsorbent systems, such as mesoporous silicas, but also to increase the stability of the amorphous form in relation to the storage time (Zůza et al., 2019). The search for better therapeutic efficacy of drugs was the main motivation for this research, using different technologies for the development of formulations with different therapeutic objectives and targets. The first part of this research aimed to develop a mucoadhesive in situ thermoreversible gel, capturing a corticosteroid for the treatment of mucous regions with inflammation. This first part produced three scientific publications that are organized as the first two chapters of this document and an annex. The second part of the research aimed to investigate the adsorbent role of mesoporous silica in two different drugs in terms of the tendency to crystallization, aiming beyond bioavailability to increase the physical stability of the systems and the impact of the drugs on the storage of the systems. Through the development of this study, a scientific article was produced presented in the third and last chapter of this document. In summary, the research developed during the PhD period is organized in two parts, in which: Chapter 01 - This chapter has mucoadhesion as its central theme, and constitutes a review of the literature published as a chapter in the book “Sciences applied to health products”, by the publisher of the State University of Goiás in 2019, ISBN 978-85-5582- 060-1 (annex 3). Strategies for the development of new mucoadhesive pharmaceutical forms, mucus and its function in the human body, the theories that analyze mucoadhesion, the mucoadhesive formulations already available and the techniques and assays used to quantify mucoadhesion were covered in this chapter. Chapter 02 - In this chapter we describe the development and characterization in vitro / in vivo of a mucoadhesive in situ gelling formulation using poloxamer 407 (Pluronic® F 127), a thermoreversible polymer, capturing budesonide (BUD), a potent corticosteroid used for treatment of a wide range of inflammatory diseases, including those that affect mucous membranes, such as in the gastrointestinal tract. This chapter was published in 2019 as an original article in the Journal of Controlled Release, entitled Thermoreversible mucoadhesive polymer-drug dispersion for sustained local delivery of budesonide to treat inflammatory disorders of the GI tract (appendix 4). The term with the approval by the ethics committee for the use of animals is attached (annex 1). Another publication, referring to oral pharmaceutical compositions of corticosteroids that gel in situ, was the production of a patent (annex 2). Patent filed and published at the United States Patent and Trademark Office, with international application under Patent Cooperation Treaty (PCT), WO 2018/193423 A1, 2018. This patent is the result of a partnership between the University and a pharmaceutical industry, Ferring Pharmaceuticals, established for the development of mucoadhesive pharmaceutical formulations. In turn, depending on the second part, which deals with technologies aimed at optimizing bioavailability and promoting greater stability using porous adsorbent systems, we have chapter 3. Chapter 03 - In this chapter, a method has been described to determine the monolayer loading capacity (MLC) of naproxen, a weak drug to amorphize, in mesoporous silica (MS). MS can be used as a carrier to stabilize the amorphous form of a drug. In addition, the impact of monolayer, pore filling and excess, on the physical stability of this system was studied and compared to ibuprofen, a strong drug to amorphize. Finally, we investigated the impact of drug loading on storage below and above the glass transition temperature (Tg), in particular with a focus on the amorphous (in) stability of the confined drug, for both drugs. Using Theoretical Functional Density Theory (DFT) and Molecular Dynamics ab initio (AIMD), the binding energies for the monolayer suggest that the monolayer is thermodynamically more favorable than the crystalline form, while the confined amorphous form is thermodynamically less favorable. This chapter was published as an original article in the International Journal of Pharmaceutics: X, in 2019 (Annex 5).Item Eletroanálise aplicada a ensaios de compatibilidade e estabilidade de fármacos(Universidade Federal de Goiás, 2020-08-31) Carvalho, Murilo Ferreira de; Garcia, Luane Ferreira; http://lattes.cnpq.br/6624146379323596; Gil, Eric de Souza; http://lattes.cnpq.br/3218622824233303; Garcia, Luane Ferreira; Leite, Karla Carneiro de Siqueira; Alecrim, Morgana Fernandes; Marreto, Ricardo Neves; Oliveira, Thiago Levi SilvaIntroduction: The pharmaceutical industry faces a major challenge to improve the solubility of some active ingredients. Carvedilol (CRV) is the drug of therapeutic choice in cardiovascular diseases. CRV presents low water solubility and low bioavaliability. Several researches were performed in order to increase the oral bioavailability, where selfemulsifying appeared as viable alternatives. Objective: The objective of this work was to evaluate interactions between CRV and its excipients, detecting the oxidative potential of these substances, thus, providing predictive data for the oxidation that may affect the stability and bioavailability of the drug system. Methodology: In the carbon pastes preparation were used 70 mg (graphite) and 30 mg (oil). Six excipients, oleic acid, canola, capmul, safflower, sesame and Plurol isoestearique. The following proportions were used in the making of the paste: 30:0, 20:10, 15:15, 10:20 and 0:30 of liquid excipient (sample): mineral oil (mg:mg) and 1.5:68.5, 3:67 and 7:63 of solid excipient (sample): graphite (mg:mg). CRV was incorporated in the oil moiety for solubilization and after used in solidstate analysis in the carbon paste. The techniques used were cyclic voltammetry, differential pulse voltammetry and electrochemical impedance spectroscopy. Results and Discussions: The Isostearique Plurol® was found to be electro-active in the studied potential, however, this oil oxidizes in different potentials from CRV. The results show that Isostearique Plurol®, liquid excipient, isoestearique acid and the solid excipient presented the greatest variations of anodic potential and were considered the best excipients for the CRV formulation. CRV presented higher stability at room temperature (25 ºC), whereas, it presented higher stability at 50 ºC when mixed with 7% estearique acid. Conclusions: Evidences of incompatibility were not found, relative to the increase of oxidation vulnerability, when in the presence of these excipients. The employment of electroanalysis for CRV compatibility studies show promising viability.Item Estudo clínico de segurança não controlado do uso de drogas vegetais em pacientes atendidos em um ambulatório público de fitoterapia(Universidade Federal de Goiás, 2020-03-02) Cirilo, Hérica Núbia Cardoso; Cunha, Luiz Carlos da; http://lattes.cnpq.br/6349547031976679; Bara, Maria Teresa Freitas; http://lattes.cnpq.br/3914164125498267; Bara, Maria Teresa Freitas; Santos, Thalyta Renata Araújo; Alves, Suzana Ferreira; Paula, José Realino de; Soares, Amanda QueirozIntroduction: The use of medicinal plants has been increasing worldwide, so as the reports of adverse events (AE) related to them. The World Health Organization Toxicity Grading Scale for Determining the Severity of Adverse Events (WHO-TGS) establishes the toxicity criteria for laboratory tests in clinical trials with humans, allowing to evaluate the safety of a treatment. Additionally, the Naranjo Algorithm establishes the causal relationship between the observed AE and the product under investigation. Main objective: To verify the safety of the use of 12 medicinal plants in subjects attended at a phytotherapy outpatient clinic of the Unified Health System, using the WHO-TGS and the Naranjo Algorithm. Methodology: This is an open and prospective uncontrolled "before and after" clinical study. The selected participants were submitted to laboratory tests before and after 30 days of use of the prescribed medicinal plants. The results were analyzed according to WHO-TGS. The medicinal plants were suspended for 30 days in those who presented laboratory alterations indicative of toxicity, the exams were repeated, and the results analyzed. The observed AE had their causality established through the Naranjo Algorithm. Results and discussion: There was a higher prevalence of female individuals, mean age of 46 years, graduated level, who sought the health service to treat depression and anxiety. The majority of them are non-users of tobacco and illicit drugs, while half regularly consume alcoholic beverages, and are characterized mainly by the daily use of medicinal plants, plant drugs and herbal medicines for therapeutic purposes. 42 subjects were selected to use the medicinal plants prescribed for 30 days, of which 17 (40.5%) did not present AE, whereas 25 (59.5%) presented Grade 1 toxicity (GT1) AE, mainly the increase in amylase concentration (n=9). After the suspension of the medicinal plants use, 14 participants continued to present GT1 AE, mainly hypomagnesemia and hematuria. The Naranjo Algorithm established a possible causal relationship between hyperamylasemia; hypomagnesemia; hematuria; AST, ALT and GGT increase; hyperglycemia; hypocalcemia; hypercalcemia; thrombocytopenia; prothrombin time elevation; proteinuria and the medicinal plants used. Conclusions: The results obtained revealed the socio-demographic, clinical, life habits and pharmacotherapeutic profile of the participants. They demonstrated the occurrence of AE after the use of Bauhinia forficata, Curcuma longa, Cynara scolymus, Equisetum arvense, Erythrina mulungu, Matricaria chamomilla, Melissa officinalis, Passiflora edulis, Phyllanthus niruri e Zingiber officinale, evidencing a possible causal relationship to the use of these medicinal plants. Such relationship corresponds to a low level of probability of occurrence of an AE by the products under investigation, that is, it demonstrates safety in the use of these medicinal plants, in the experimental conditions employed.Item Preparação e uso de eletrodos de frafite nanoestruturados de vanadato de bismuto na degradação fotoeletrocatalítica de radamina B isotiacianato(Universidade Federal de Goiás, 2023-02-10) Isecke, Bruna Guimarães; Teixeira, Guilhermina Ferreira; http://lattes.cnpq.br/5992362464264556; Gil, Eric de Souza; http://lattes.cnpq.br/3218622824233303; Gil, Eric de Souza; Oliveira, Sérgio Botelho de; Colmati Júnior, Flávio; Oliveira, Mayk Teles de; Prado, Lara Barroso BritoPhotoelectrocatalysis is a promising way to treat water contaminated by negative organic compounds. The use of BiVO4 nanoparticles supported on a conductive substrate allows the degradation of pollutants to be carried out under less energetic conditions. In this work, we report the degradation of Rhodamine B Isothiocyanto dye in a BiVO4/graphite (BVO@C) electrode performed under visible light irradiation through a photoelectrocatalytic process. The BiVO4 particles passed through ultrasonic irradiation by coprecipitation and then deposited on the surface of a graphite by the impregnation method. The 23 factorial design was used to optimize electrode fabrication. The electrode was characterized using SEM/EDS, XRD and DLS techniques. For the electrochemical characterization of the electrode, cyclic voltammetry (VC) and electronic impedance spectroscopy (EIS) were performed. The evaluation of the degradation of RhB isothionate was carried out in the processes of electrooxidation and photoelectrocatalysis, controlling or not controlling the temperature using the unmodified electrode and the BVO@C electrode. The characterization of the electrode confirmed the impregnation of the graphite with a BiVO4 particle through SEM/EDS, revealing specific peaks referring to vanadium (V), bismuth (Bi) and oxygen (O). The XRD analysis bought peaks related to the monoclinic phase of BiVO4, this fact being important due to the smaller band gap value, which implies activation of the semiconductor through light in the visible range. DLS and zeta potential analysis revealed negative charges on the surface of BVO@C, PDI values that indicated that the solution used for impregnation had monodispersive characteristics, which corroborated to a less regular impregnation, since this factor does not contribute to a deposition independent. Electrochemical analyzes confirm a considerable improvement of the modified electrode through a decrease in resistivity that may be associated with the presence of BiVO4, in addition to a decrease in hysteresis in relation to the emission/reduction pair. After 30 minutes, the degradation results in the electrocatalysis (EC) and photoelectrocatalysis (PEC) system, the results were considered relevant, reaching almost 100% of the dye being degraded, with a higher degradation rate for the photoelectrocatalysis experiments in relation to electroxidation, and higher values in experiments in which temperature was not controlled. To analyze the electrode reuse cycles, several reuse cycles were performed and the electrode efficiency (BVO@C) was observed. After nine cycles of reuse of the BVO@C electrode, it was observed that it presented more than 80% of RhB efficiency. and photoelectrocatalysis of dyes from industrial effluents.Item Avaliação do potencial antioxidante e atividade vasorrelaxante de Cinnamomum sp.(Universidade Federal de Goiás, 2023-05-16) Moreno, Emily Kussmaul Gonçalves; Macêdo, Isaac Yves Lopes de; http://lattes.cnpq.br/7801802836007488; Gil, Eric de Souza; http://lattes.cnpq.br/3218622824233303; Macêdo, Isaac Yves Lopes de; Garcia, Telma Alves; Rocha, Matheus Lavorenti; Carvalho, Murilo Ferreira de; Machado, Fabio BahlsThe great interest in the study of antioxidants is mainly the prevention of physiological damage caused by oxidative processes. Cinnamon (Cinnamomum sp.), is a spice used worldwide in cooking and has gained prominence for its pharmacological properties, such as anti-inflamatory, antibacterial, antifungal, cardiovascular, among others. Given the importance of determining the antioxidant capacity in relation to therapeutic methods, this study aims to evaluate the antioxidant profile of commercial cinnamon samples by spectrophotometric, electrochemical and vasorelaxant methods. The spectrophotometric methods performed were 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2’-azinobis (3- ethylbenzothiazoline-6-sulfonic acid) (ABTS) and Folin-Ciocalteu. For the electrochemical experiments, a three-electrode system was used, consisting of carbon paste electrode, platinum wire, and Ag/AgCl/KClsat, representing the working, auxiliary, and reference electrodes, respectively. The electroanalytical methods used were differential pulse, square wave, and cyclic voltammetries. Vascular reactivity was evaluated in aortic artery rings from male Wistar rats, using arteries with intact vascular endothelium and without endothelium. The endothelial nitric oxide synthase (eNOS) pathway and the inhibitor N-nitro-L-arginine-methylester (L-NAME) were used in this analysis. The correlation between the electrochemical approach and total phenols by the ABTS, DPPH, and Folin- Ciocalceu methods were 0.63, 0.70, and 0.73, respectively, with 1 being an ideal correlation. The correlation between spectrophotometric methods was 0.83. For the electrochemical results, a similar profile was observed, with anodic peak a.c Epa1= 0.1 V, indicative of polyphenolic compounds with high antioxidant power. In addition, a biosensor on a carbon paste electrode was applied to the selected extracts using laccase enzyme, obtained by the fungus Marasmiellus sp. and a signal improvement of up to 4 times in the antioxidant profile was observed when compared to the electrode without modification. The samples were analyzed by mass spectrometer, and the main chemical markers found were coumarin, cinnamaldehyde, and eugenol. Pharmacological assays showed that these samples also promoted a significant vasorelaxant effect associated to the antioxidant potential. Thus, cinnamon showed a high antioxidant capacity, in agreement with the results obtained in other studies, emphasizing its importance as a functional food.Item Os estágios iniciais do desenvolvimento do zebrafish como modelo alternativo para predizer a toxicidade oral aguda in vivo(Universidade Federal de Goiás, 2021-02-11) Prado, Lara Barroso Brito; Valadares, Marize Campos; http://lattes.cnpq.br/6157755243167018; Oliveira, Gisele Augusto Rodrigues de; http://lattes.cnpq.br/6221735418479539; Oliveira, Danielle Palma de; Rocha, Matheus Lavorenti; Ferreira, Monica Valdyrce dos Anjos Lopes; Alves, Vinícius de Medeiros; Valadares, Marize CamposZebrafish (Danio rerio) early-life stages offer a complex and multicellular system integrating various tissues and differentiation processes. In addition, the zebrafish embryos are structurally and functionally similar to vertebrates, including humans. The acute toxicity test with zebrafish embryos and larvae is already worldwide recognized as an alternative model for ecotoxicological assessment, but it is not include in normative of the Council fot the Control of Animal Experimentation (CONCEA), wich recognize the use of alternative methods validated in research activities in Brazil. This organism model can filling the gap between conventional in vitro and in vivo tests for extrapolation of data for humans, the present study aimed to assess the acute toxicity of substances with different Global Harmonization System (GHS) categories using zebrafish early-life stage to determine LC50 values and compared with in vivo (LD50) acute oral toxicity data from literature, in order to generate a model to prediction acute oral toxicity. This prediction model was evaluated by the application of a drug candidate (LQFM 021). Fifteen substances were evaluated by Fish Embryo Acute Toxicity (FET) test (OECD 236). Parameter evaluated was lethal and sublethal effects after 96 h of exposure. A linear regression- model using the log-transformed of the LC50 values and LD50 was generated for the estimated of LD50 from LC50 values. This model resulted in the following equation Log LD50 (mg/kg) = 0.5749 x log LC50 (mg/L) + 1.284. The method domain of application was 53.33% and the R2 was 0,66. Sublethal effects indicate that substances more toxic presented more abnormalities. The DL50 predicted with FET testing LQFM 021, which classified as category 4 in GHS acute oral systemic toxicity assessment, was of 408.52 mg/kg, which also classified as Category 4. In this work, Quantitative Activity-Activity Relationship (QAAR) models were also developed, based on the data obtained in the FET test with zebrafish. This model using seven toxicological descriptors generated statistically predictive models with R2 values ranging from 0.80 to 0.95 and in combination with the Random Forest method it presented the best performance in the prediction of acute oral toxicity in vivo. Therefore, our results suggest that early-life stages of zebrafish could be at least a refinement in the sense of the principles of the 3R’s to predict acute oral toxicity in vivo, being as an intermediary in the preclinical evaluation between in vitro and in vitro.Item Imunossensor impedimétrico para determinação de rituximabe aplicado ao controle qualidade e monitoramento clínico(Universidade Federal de Goiás, 2023-02-10) Rodrigues, Edson Silvio Batista; Neves, Bruno Júnior; http://lattes.cnpq.br/7256565904920282; Gil, Eric de Souza; http://lattes.cnpq.br/3218622824233303; Gil, Eric de Souza; Sgobbi, Lívia Flório; Feres, Valeria Christina De Rezende; Oliveira Neto, Jerônimo Raimundo de; Ferreira Júnior, ÁlvaroIntroduction: Cancer is an umbrella term for a large group of diseases that can affect any part of the body. Rituximab (RTX) is a chimeric monoclonal antibody (mAb) formed by the mouse/human combination, which specifically binds to the CD20 cell marker that is expressed in normal and malignant B lymphocytes, exerting significant antitumor activity. In this context, RTX emerged as a therapeutic alternative in the treatment of various pathologies, including autoimmune diseases, inflammatory diseases and various types of cancer in which sensitive and low-cost methods are required for therapeutic drug monitoring (TDM) and quality control (QC) of this drug. Objectives: To develop a selective electrochemical immunosensor for the detection and quantification of RTX in urine and blood. Methodology: The ploliconal antibody (pAb) was purified from chicken egg, used as a biorecognition element to create an immunosensor for the detection and quantification of RTX. The immunosensor was constructed from the immobilization of the RTX IgY antibody on the glassy carbon electrode surfaces. The characterization of the immunosensor was obtained through monitoring at each stage via cyclic voltammetry and electrochemical impedance spectroscopy, where selectivity is obtained by specifically binding pAb and RTX and casein is used to block possible nonspecific interactions of other proteins. After the development of the immunosensor, the validation of the method was carried out, evaluating the selectivity, accuracy, precision, linearity and limits of detection (LOD) and quantification (LOQ). Finally, the RTX was determined in urine and blood samples. Results and discussion: Linearity was demonstrated between charge transfer resistance and RTX from 2 to 14 μg mL−1 (r2 of 0.99), with LOD and LOQ of 130 and 400 ng mL−1 respectively. The method showed adequate precision with a RPD of 0.96% for researcher 1 and 1.27% for researcher 2, and accuracy with a RPD of 1.30%, with a confidence interval between 99.0% and 101.0 %. Selectivity was achieved by demonstrating the binding specificity between the antibody and RTX, where casein blocks non-specific binding when the immunosensor is applied to blood and urine samples. Conclusion: The developed immunosensor presented precision and accuracy for the detection of RTX with results in approximately 20 minutes. In this way, low-cost, marker-sensitive electrochemical immunosensors can help in TDM, QC and extended stability monitoring of different drugs, in a simple way and with versatile assays.Item Avaliação toxicológica de uma formulação a base de glifosato e seus principais constituintes sobre o estágio embrio-larval de zebrafish: exposição única e repetida(Universidade Federal de Goiás, 2021-06-12) Rodrigues, Laís de Brito; Oliveira, Gisele Augusto Rodrigues de; http://lattes.cnpq.br/6221735418479539; Oliveira, Gisele Augusto Rodrigues de; Fil, Eric de Souza; Valadares, Marize Campos; Rocha, Thiago Lopes; Sabóia-Morais, Simone Maria Teixeira deGlyphosate (GLY) is the active ingredient of several herbicide formulations used to control weeds in agricultural and non-agricultural areas. Due to intensive use of GLY-based formulations and the repeated applications once weed resistance, some of their components, including the active ingredient, may reach the aquatic environment through direct run-off and leaching. The present study assessed the acute toxicity, after continuous and repeated exposures, and genotoxicity of the GLY-based formulation Atanor 48 (ATN) and its major constituents GLY, surfactant polyethoxylated tallow amine (POEA), as well as the main metabolite of GLY aminomethylphosphonic acid (AMPA) on zebrafish early life stages. Also, we evaluate larvae resilience after ATN, GLY and POEA pulsed-exposure. The toxic effects of these chemicals were evaluated in the fish embryo acute toxicity test with zebrafish (Danio rerio), while genotoxic effects were investigated in the comet assay with cells from zebrafish larvae and rainbow trout gonad-2 (RTG-2). GLY and AMPA caused no acute toxic effect after continuous exposure, while ATN and POEA induced significant lethal effects in zebrafish (LC50-96 h 76.50 mg/L and 5.49 mg/L, respectively. In summary, these data indicate that the lethal effects on zebrafish early-life stages can be ranked in the following order from most to least toxic: POEA > ATN > GLY ≈ AMPA. All compounds were genotoxic to zebrafish larvae (LOEC 1.7 mg/L for GLY, ATN, AMPA and 0.4 mg/L for POEA). Unlike in vivo. POEA induced DNA damage in RTG-2 cells (LOEC 1.6 mg/L), suggesting that it is a direct acting genotoxic agent. GLY caused no acute toxic effect after repeated pulse exposure. However, ATN showed significant mortality (LC50-96 h 148.80 mg/L) after 5 h pulse at 100 mg/L and POEA induced significant toxicity on zebrafish early life stages after 1, 2 and 5 h pulse with LC50-96 h of 43.49 mg/L, 47.23 mg/L and 11.61 mg/L, respectively. Zebrafish was not able to reverse the sublethal effects induced by ATN, GLY and POEA during the recovery period. The toxic effects induced by ATN and POEA after pulsed-exposure were less than continuous exposure. Therefore, its important to evaluate different toxicological endpoints with distinct exposure scenarios to predict the hazards of GLY-based formulations, their components and breakdown product to aquatic biota.Item Estratégias de bioengenharia e Novas Abordagens Metodológicas (NAMs) aplicadas à avaliação do potencial de sensibilização pulmonar de toxicantes inalados(Universidade Federal de Goiás, 2022-11-10) Silva, Artur Christian Garcia da; Valadares, Marize Campos; http://lattes.cnpq.br/6157755243167018; Valadares, Marize Campos; Cunha, Luiz Carlos da; Fonseca, Simone Gonçalves da; Leite, Jacqueline Alves; Bakuzis, Andris FigueiroaIntroduction: Respiratory sensitization encompasses a group of inflammatory diseases that manifest through airway hyperresponsiveness and airflow limitation. Although the chemical respiratory allergy (CRA) induced by Low Molecular Weight (LMW) sensitizers is a major concern, especially in terms of the regulatory framework, to date there are no methods available for preclinically addressing this toxicological outcome, as its mechanistic background is not fully understood at molecular or cellular levels. Objectives: The objective of this work is to apply several in vitro and in chemico approaches in order to address physiologically relevant aspects of human response to LMW respiratory allergens, aiming to contribute to the elucidation of this toxicant class mode of action and further evaluation regarding the preclinical stage. Methodology: First, we evaluated the interaction between LMW sensitizers and lung mucus by employing the mucin spectroscopic profile assessment in the presence and absence of seven chemical respiratory allergens. Then, we performed the evaluation of inflammatory and functional parameters in different respiratory tract cell types, including bronchial epithelial (BEAS-2B), lung fibroblasts (MRC-5), endothelial (EA.hy926) and monocytic cells (THP-1), after exposure to sub-cytotoxic concentrations of each sensitizer. Following the monolayer cell-based studies, we integrated the four mentioned cell tupes in a 3D bronchial co-culture model and exposed it to maleic anhydride aerosols in an air-liquid interface (ALI) for further evaluating inflammatory and functional tissue parameters. Finally, we also proposed the development of a bronchial epithelial model using the porcine descellularized bronchial wall as a bioscaffold, for which we evaluated different obtention methods, as well as the cell behavior when cultivated upon the obtained matrix. Results/Discussion: The results showed that some of the sensitizers evaluated interact with mucin, the main protein mucus component, but the toxicant-mucin complex formation does not seem to be a common feature of different chemical classes of allergens. At a cellular level, sensitizers promoted an increase in IL-8, IL-6, and IL-1β production in terms of the different evaluated cell types. It also impaired the MUC1 expression by bronchial cells BEAS-2B and activated endothelial cells (EA.hy926), thereby increasing the ICAM-1 surface detection. It has been also demonstrated an increased expression of dendritic cell activation/maturation surface biomarkers (CD86, HLA-DR and CD11b) in THP-1 monocytic cells after exposure to chemical allergens. In parallel, the 3D bronchial co-culture model was successfully reconstructed using by cultivating the four aforementioned cell types at an ALI, and maleic anhydride aerosols exposure increased the inflammatory biomarkers production, as well as the apoptosis in epithelial layer and THP-1 dendritic cell activation. Besides, the decellularized porcine bronchial matrix allowed the cultivation of Calu-3 cells, which regularly expressed the airway epithelium biomarkers after 7 and 14 days of cultivation in ALI. Conclusion: Taken together, our results showed that these aforementioned cell types participate in the CRA Adverse Outcome Pathway and must be considered when developing testing strategies. The integration of different cell types in an unique co-culture model allowed the obtention of a global toxicant response, enabling the aerosol exposure, which emulates more realistically the airway contact with external aggressors. Finally, the 3D bronchial bioscaffold yielded the obtention of an epithelial model that morphologically and phenotypically resembles human airway epithelium, being a useful alternative for addressing lung response to inhaled toxicants. Thus, the work subsidizes the employment of inflammatory and functional biomarkers, as well as of bioengineering-derived in vitro models for addressing the respiratory sensitization potential of LMW allergens at a preclinical stage.Item Avaliação toxicológica de protótipos à base de rutênio para o tratamento do câncer utilizando metodologias alternativas ao uso de mamíferos(Universidade Federal de Goiás, 2020-09-09) Teixeira, Thallita Monteiro; Lacerda, Elisângela de Paula Silveira; http://lattes.cnpq.br/9390789693192751; Lacerda, Elisângela de Paula Silveira; Grisólia, César Koppe; Machado, Mônica Rodrigues Ferreira; Sá, Viviane de Souza Velozo; Franchi, Leonardo PereiraAufgrund der weltweit zunehmenden Zahl von Krebsfällen haben sich Rutheniumkomplexe als vielversprechende Chemotherapeutika erwiesen, da sie eine geringe Toxizität aufweisen und selektiver für Tumorgewebe sind. Die RuLap- und RuLau-Rutheniumkomplexe wurden gegen mehrere Tumor- und Nicht-Tumorzelllinien getestet, um die In-vitro-Zytotoxizität durch den MTT-Test und auch Neutralrot zu bewerten. Der RuLau-Komplex zeigte einen Selektivitätsindex von 1,17 bis 10,91 für die HeLa-Tumorlinie. Bei der Auswertung des In-vitro-Mincronukleus-Tests war die RuLau-Verbindung diejenige mit dem niedrigsten genotoxischen Potential, die nur bei einer Konzentration von 2 mg / l eine signifikante Genotoxizität zeigte. Im Kometen-In-vitro-Assay zeigten sowohl RuLap- als auch Rulau-Verbindungen eine genotoxische Wirkung Potenzial, ohne Unterschied zwischen den Belichtungszeiten von 24 Stunden und 48 Stunden. In akuten Toxizitätstests an Zebrafischembryonen zeigte die RuLau-Verbindung im Vergleich zur RuLap-Verbindung weniger ausgeprägte subletale Wirkungen. Die Rulau-Verbindung ließ die Embryonen selbst bei den höchsten Konzentrationen nicht sterben. Die RuLap-Verbindung war die einzige Verbindung, die tatsächlich toxischer war, was zu einer Verzögerung des Schlupfes der Embryonen führte und auch deren Letalität verursachte, noch bevor sie schlüpften. Im neurotoxischen Potentialbewertungstest unter Verwendung des Netzhauttests bei Zebrafischembryonen gab es bei den niedrigsten Konzentrationen (½ EC10 und EC10) keine signifikante Abnahme im Bereich der inneren plexiformen Schicht (CPI), was nur bei der höchsten Konzentration Veränderungen zeigte (2x EC10), und als die Netzhautdicke bewertet wurde, gab es eine signifikante Zunahme der Netzhautdicke bei der niedrigsten Konzentration. Im Acetylcholinesterase-Hemmungstest induzierte keine der getesteten Verbindungen die Enzymhemmung, nicht einmal die RuLau-Verbindung, die möglicherweise mit der Nichtinduktion neurotoxischer Wirkungen zusammenhängt. Der RuLau-Rutheniumkomplex zeigt dann eine größere Selektivität für Tumorstämme und ein geringeres toxisches Potential, wenn er gegen Zebrafischembryonen bewertet wird.