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Zidovudine (AZT) has been widely used alone or in combination with other antiretroviral drugs for the treatment of human immunodeficiency virus. Its erratic oral bioavailability necessitates frequent administration of high doses, resulting in severe side effects. In this study, the design of mucoadhesive solid dispersions (SDs) based on chitosan (CS) and hypromellose phthalate (HP) was rationalized as a potential approach to modulate AZT physicochemical and pharmaceutical properties. SDs were prepared at different drug:polymer ratios, using an eco-friendly technique, which avoids the use of organic solvents. Particles with diameter from 56 to 73 µm and negative zeta potentials (-27 to -32 mV) were successfully prepared, achieving high drug content. Infrared spectroscopy revealed interactions between polymers but no interactions between the polymers and AZT. Calorimetry and X-ray diffraction analyses showed that AZT was amorphized into the SDs. The mucoadhesive properties of SDs were evidenced, and the control of AZT release rates from the matrix was achieved, mainly in acid media. The simple, low-cost, and scalable technology proposed for production of SDs as a carrier platform for AZT is an innovative approach, and it proved to be a feasible strategy for modulation the physico-chemical, mucoadhesive, and release properties of the drug.

COVID-19 has resulted in more than 490 million people being infected worldwide, with over 6 million deaths by April 05th, 2022. Even though the development of safe vaccine options is an important step to reduce viral transmission and disease progression, COVID-19 cases will continue to occur, and for those cases, efficient treatment remains to be developed. Here, a drug repurposing strategy using nanotechnology is explored to develop a therapy for COVID-19 treatment. Nanoparticles (NPs) based on PLGA for fingolimod (FTY720) encapsulation show a size of ∼150 nm and high drug entrapment (∼90%). The NP (NP@FTY720) can control FTY720 release in a pH-dependent manner. Cytotoxicity assays using different cell lines show that NP@FTY720 displays less toxicity than the free drug. Flow cytometry and confocal microscopy reveal that NPs are actively internalized mostly through caveolin-mediated endocytosis and macropinocytosis pathways and co-localized with lysosomes. Finally, NP@FTY720 not only exhibits anti-SARS-CoV-2 activity at non-cytotoxic concentrations, but its biological potential for viral infection inhibition is nearly 70 times higher than that of free drug treatment. Based on these findings, the combination of drug repurposing and nanotechnology as NP@FTY720 is presented for the first time and represents a promising frontline in the fight against COVID-19.

Made available in DSpace on 2022-05-01T08:15:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-11-05 Universidade Estadual Paulista Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Universidade de São Paulo Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Nanotechnology has been widely used in the development of polymer nanocarriers for the oral delivery of biomolecules. The use of nanostructured systems can improve drug delivery and help to overcome the disadvantages of the oral administration of peptides. Nanoparticles (NPs) based on gellan gum (GG) and chitosan (CS) blends were prepared through polyelectrolyte complexation. The influence of pH on the zeta potential (ZP) of polymers allowed the selection of pH 5.0 as the most suitable pH for the complexation of polyelectrolytes. The effects of the polymer mass ratio and addition order on the formation and physicochemical properties of the NPs were evaluated. All NPs showed high positive ZP (> + 30 mV), which ensures electrostatic stability. The order of addition of the polymers influenced the particle size. Nanoscale structures (575.30–974.60 nm) were formed when GG (0.5–3 mg) was dripped into the CS dispersion (0.75–4.5 mg); however, when CS was dripped in the GG dispersion, particle aggregation occurred (sizes >5000 nm). Polymyxin B (PMB) nanoencapsulation reduced the particle size, mainly at low GG mass (1.5 mg). The PMB–polymer interactions were detected by Fourier transform infrared spectroscopy. The X-ray diffraction data indicated the formation of more organized structures with a higher degree of crystallinity. Scanning electron microscopy revealed spherical and uniform NPs. The mucoadhesive capability of the NPs was also demonstrated. The NP2_1 and NP2_3 released the lowest amount of drug in HCl 0.1 N (pH 1.2) (

Made available in DSpace on 2019-10-06T15:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-04-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Fundació Catalana de Trasplantament European Regional Development Fund Alginate-based polyelectrolyte complexes (PECs) and hydrogel were engineered as platforms for local bevacizumab (BVZ) therapy. This study provides deep comprehension on the microstructures of such systems, and their correlation with drug-release patterns. PECs and hydrogel were characterized using Fourier transform infrared spectroscopy, small-angle X-ray scattering, scanning electron microscopy, atomic force microscopy, and porosimetry. Structural investigations indicated that PECs are formed by supramolecular interactions, resulting in physically cross-linked polymer networks, whereas the BVZ-loaded hydrogel has a more compact and rigid structure, promoting better entrapment of BVZ. PECs and hydrogel were able to control the BVZ release for 4 and 8 days, respectively. Their release profiles correlated best with the Higuchi and Korsmeyer-Peppas models, respectively, indicating drug diffusion as the limiting step for drug release. Furthermore, BVZ remained biologically active in vitro after its incorporation into the hydrogel system. Together, these studies confirm that PECs and hydrogel exhibit different porous structures and physicochemical properties, making them promising platforms that allow the modulation of BVZ release meeting different requirements. Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista (UNESP), Araraquara, Rodovia Araraquara–Jaú km 1 I3S–Instituto de Investigação e Inovação em Saúde Universidade do Porto, Rua Alfredo Allen, 208 INEB–Instituto de Engenharia Biomédica Universidade do Porto, Rua Alfredo Allen, 208 CESPU–Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde, Rua Central de Gandra 1317 ICBAS–Instituto Ciências Biomédicas Abel Salazar Universidade do Porto, Rua de Jorge Viterbo Ferreira 228 Instituto de Química Universidade Estadual Paulista (UNESP), Araraquara, Rua Professor Francisco Degni, 55 Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista (UNESP), Araraquara, Rodovia Araraquara–Jaú km 1 Instituto de Química Universidade Estadual Paulista (UNESP), Araraquara, Rua Professor Francisco Degni, 55

Made available in DSpace on 2021-06-25T10:31:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-06-15 Mutations on the epidermal growth factor receptor (EGFR), induction of angiogenesis, and reprogramming cellular energetics are all biological features acquired by tumor cells during tumor development, and also known as the hallmarks of cancer. Targeted therapies that combine drugs that are capable of acting against such concepts are of great interest, since they can potentially improve the therapeutic efficacy of treatments of complex pathologies, such as glioblastoma (GBM). However, the anatomical location and biological behavior of this neoplasm imposes great challenges for targeted therapies. A novel strategy that combines alpha-cyano-4-hydroxycinnamic acid (CHC) with the monoclonal antibody cetuximab (CTX), both carried onto a nanotechnology-based delivery system, is herein proposed for GBM treatment via nose-to-brain delivery. The biological performance of Poly (D,L-lactic-co-glycolic acid)/chitosan nanoparticles (NP), loaded with CHC, and conjugated with CTX by covalent bonds (conjugated NP) were extensively investigated. The NP platforms were able to control CHC release, indicating that drug release was driven by the Weibull model. An ex vivo study with nasal porcine mucosa demonstrated the capability of these systems to promote CHC and CTX permeation. Blot analysis confirmed that CTX, covalently associated to NP, impairs EGRF activation. The chicken chorioallantoic membrane assay demonstrated a trend of tumor reduction when conjugated NP were employed. Finally, images acquired by fluorescence tomography evidenced that the developed nanoplatform was effective in enabling nose-to-brain transport upon nasal administration. In conclusion, the developed delivery system exhibited suitability as an effective novel co-delivery approaches for GBM treatment upon intranasal administration. School of Pharmaceutical Science São Paulo State University UNESP, Rodovia Araraquara/Jaú Km 01 Laboratório de Nanotecnologia Farmacêutica e Sistemas de Liberação de Fármacos FarmaTec Faculdade de Farmácia Universidade Federal de Goiás - UFG, 5ª Avenida c/Rua 240 s/n, Praça Universitária Life and Health Sciences Research Institute (ICVS) School of Medicine University of Minho ICVS/3B's-PT Government Associate Laboratory Molecular Oncology Research Center Barretos Cancer Hospital School of Pharmaceutical Science São Paulo State University UNESP, Rodovia Araraquara/Jaú Km 01

Made available in DSpace on 2021-06-25T11:13:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-03-01 Polymer blends of gellan gum (GG)/retrograded starch(RS) and GG/pectin (P) were cross-linked with calcium, aluminum, or both to prepare mucoadhesive microparticles as oral carriers of drugs or nano systems. Cross-linking with different cations promoted different effects on each blend, which can potentially be explored as novel strategies for modulating physical-chemical and mucoadhesive properties of microparticles. Particles exhibited spherical shapes, diameters from 888 to 1764 µm, and span index values lower than 0.5. Blends of GG: P cross-linked with aluminum resulted in smaller particles than those obtained by calcium cross-linking. GG: RS particles exhibited larger sizes, but cross-linking this blend with calcium promoted diameter reduction. The uptake rates of acid medium were lower than phosphate buffer (pH 6.8), especially GG: RS based particles cross-linked with calcium. On the other hand, particles based on GG: P cross-linked with calcium absorbed the highest volume of acid medium. The percentage of systems erosion was higher in acid medium, but apparently occurred in the outermost layer of the particle. In pH 6.8, erosion was lower, but caused expressive swelling of the matrixes. Calcium cross-linking of GG: RS promoted a significantly reduction on enzymatic degradation at both pH 1.2 and 6.8, which is a promising feature that can provide drug protection against premature degradation in the stomach. In contrast, GG: P microparticles cross-linked with calcium suffered high degradation at both pH values, an advantageous feature for quickly releasing drugs at different sites of the gastrointestinal tract. The high mucoadhesive ability of the microparticles was evidenced at both pH values, and the Freundlich parameters indicated stronger particle-mucin interactions at pH 6.8. School of Pharmaceutical Science São Paulo State University (UNESP) Araraquara, Road Araraquara-Jaú, Km 01 School of Pharmaceutical Science São Paulo State University (UNESP) Araraquara, Road Araraquara-Jaú, Km 01

Nanotechnology and nanomaterials have rapidly evolved and are used in a wide range of areas. When dealing with products that are subject to health regulations, nanotechnology-based products face a great challenge. Although nanotechnology-based products are already commercially available, there are currently no unified international guidelines, and most respected regulatory agencies consider different aspects. Furthermore, a few important gaps in knowledge still need to be filled so that safety and efficiency can be guaranteed in all nanotechnology-based products that require sanitary regulation. However, even after efficiency and safety are well-defined, significant challenges associated with the conception and operation of production processes can be directly linked to the final quality of these products. In face of the diversity and complexity of these products, a regulatory evaluation must be based on profound scientific knowledge and must assess potential risks. In this scenario, the Quality by Design (QbD) and Quality Risk Management (QRM) methods can synergistically contribute to elaborating guidelines that are in compliance with regulatory demands. The first method, QbD, is a systemic approach to product development and is based on predefined objectives. The second, QRM, is a systematic process for evaluating, controlling, and inspecting product quality throughout its pharmaceutical life cycle. Applying the elements of QbD and QRM systemizes acquisition of knowledge by scientific approach and uses elements of risks and opportunities throughout the process, which is especially important when developing products based on nanotechnology which are subject to sanitary regulation. In this chapter, we will discuss the international regulatory guidelines adopted for nanotechnology-based products that require sanitary regulation. We will also analyze QbD and QRM methodologies, which can be adopted when developing such products. In order to contextualize the theoretical discussion, this chapter will also present examples of how these guidelines have been applied. Case studies of commercially available nanotechnology-based products subject to the regulatory health approach will be also be presented and analyzed.

Made available in DSpace on 2022-04-29T08:32:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-12-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The structural fragility of monoclonal antibodies (mAbs), such as bevacizumab (BVZ), is a critical parameter for oral administration and can limit the use of several technologies to produce oral nanocarriers for these biomolecules. Nanoparticles (NPs) based on gellan gum (GG) and retrograded starch (RS) were rationally designed through polyelectrolyte complexation, and ionic cross-linking was exploited as an additional technological strategy to modulate the properties of nanocarriers. According to static light scattering analysis, the molecular weights of GG and RS were approximately 158 kDa and 1803 kDa, respectively. The influence of pH on the zeta potential (ZP) of polymers allowed the selection of pH 6.2 as the most suitable pH for the complexation of these polyelectrolytes. Non-cross-linked NPs were prepared at different polymer:drug ratios and the effects of formulation variables (polyelectrolyte ratio, drug and cross-linker concentrations, and polymer:drug ratio) on the formation and properties (size, ZP, and PDI) of cross-linked NPs were evaluated using a 33 full-factorial design. The average size of non-cross-linked and cross-linked NPs ranged from 260.1 - 299.6 nm to 265.7–629.9 nm, respectively. NPs-negative ZP (>- 20 mV) and high association efficiency (AE%) (>56.16%) were achieved. Cross-linking significantly increased the BVZ AE% (85%–100%). Analyses by attenuated total reflectance-Fourier transform infrared, fluorescence, circular dichroism, and differential scanning microcalorimetry techniques demonstrated that polyelectrolyte complexation and ionic cross-linking did not denature the secondary and tertiary structures of BVZ. The results revealed the suitability of the technological approaches used to produce BVZ-loaded nanocarriers with tailored properties, representing a potential platform for the oral delivery of BVZ. UNESP - São Paulo State University School of Pharmaceutical Sciences Department of Drugs and Medicines UNESP - São Paulo State University Institute of Chemistry Department of Analytical Chemistry Physical Chemistry and Inorganic USP - São Paulo University São Carlos Institute of Chemistry Department of Chemistry and Molecular Physics UNESP - São Paulo State University School of Pharmaceutical Sciences Department of Drugs and Medicines UNESP - São Paulo State University Institute of Chemistry Department of Analytical Chemistry Physical Chemistry and Inorganic CAPES: 001 FAPESP: 2014/50928-2 FAPESP: 2015/21412-0 FAPESP: 2017/16324-0 CNPq: 465687/2014-8

Made available in DSpace on 2021-06-25T10:18:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-04-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Financiadora de Estudos e Projetos Fundação de Amparo à Pesquisa do Estado de Goiás The development of rational therapies against complex diseases, such as cancer, has increased in the past few years due to the advances of ‘omics’ technologies. Concomitantly, several efforts have been made to design sophisticated drug delivery systems in order to increase specificity and drug accumulation in tumor sites. The complexity of these drug delivery systems highlights the need for suitable analytical methods to determine encapsulation/conjugation efficiency of drugs and molecules responsible for the targeted delivery. Therefore, this study focuses on the development and validation of a RP-HPLC-DAD methodology for concurrent quantification of paclitaxel (PTX) and cetuximab (CTX) in immunoliposomes. Chromatographic separation was achieved using a wide pore C8 column, and a gradient mobile phase consisting of 0.1% trifluoroacetic acid (TFA) in Milli-Q water/acetonitrile/isopropanol with a flow rate of 1 mL min−1. Drug peaks were fully separated and detected at 280 nm using UV detector. The method was validated according to ICH and FDA guidelines in terms of specificity and forced degradation studies, system suitability, linearity, limit of detection, limit of quantification, repeatability, intermediate precision, accuracy, robustness, and short-term stability. The developed method was linear over the concentration range of 37.5–150 μg mL−1 of PTX and 75–300 μg mL−1 of CTX. All parameters evaluated satisfied the acceptance criteria, according to both FDA and ICH guidelines. The applicability of the analytical method was assessed following the development of PTX-loaded immunoliposomes conjugated with CTX. Thus, the present study shows a novel, simple, stability-indicating and suitable method to quantify simultaneously PTX and CTX in immunoliposomes. Laboratory of Pharmaceutical Nanotechnology and Drug Delivery Systems – FarmaTec School of Pharmacy Federal University of Goiás School of Pharmaceutical Sciences São Paulo State University UNESP School of Pharmaceutical Sciences São Paulo State University UNESP Financiadora de Estudos e Projetos: #2016/09671-3 Financiadora de Estudos e Projetos: #2018/04546-1 Fundação de Amparo à Pesquisa do Estado de Goiás: 201710267000059 Fundação de Amparo à Pesquisa do Estado de Goiás: DFX2018081000125

Made available in DSpace on 2020-12-12T01:31:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 Nose-to-brain delivery is a promising approach to target drugs into the brain, avoiding the blood-brain barrier and other drawbacks related to systemic absorption, and enabling an effective and safer treatment of diseases such as glioblastoma (GBM). Innovative materials and technologies that improve residence time in the nasal cavity and modulate biological interactions represent a great advance in this field. Mucoadhesive nanoparticles (NPs) based on poly(lactic-co-glycolic acid) (PLGA) and oligomeric chitosan (OCS) were designed as a rational strategy and potential platform to co-deliver alpha-cyano-4-hydroxycinnamic acid (CHC) and the monoclonal antibody cetuximab (CTX) into the brain, by nasal administration. The influence of formulation and process variables (O/Aq volume ratio, Pluronic concentration, PLGA concentration, and sonication time) on the properties of CHC-loaded NPs (size, zeta potential, PDI and entrapment efficiency) was investigated by a two-level full factorial design (24). Round, stable nano-sized particles (213–875 nm) with high positive surface charge (+ 33.2 to + 58.9 mV) and entrapment efficiency (75.69 to 93.23%) were produced by the emulsification/evaporation technique. Optimal process conditions were rationally selected based on a set of critical NP attributes (258 nm, + 37 mV, and 88% EE) for further conjugation with CTX. The high cytotoxicity of CHC-loaded NPs and conjugated NPs was evidenced for different glioma cell lines (U251 and SW1088). A chicken chorioallantoic membrane assay highlighted the expressive antiangiogenic activity of CHC-loaded NPs, which was enhanced for conjugated NPs. The findings of this work demonstrated the potential of this nanostructured polymeric platform to become a novel therapeutic alternative for GBM treatment. [Figure not available: see fulltext.]. School of Pharmaceutical Science São Paulo State University UNESP, Rodovia Araraquara/Jaú Km 01 Life and Health Sciences Research Institute (ICVS) School of Medicine University of Minho ICVS/3B’s-PT Government Associate Laboratory Molecular Oncology Research Center Barretos Cancer Hospital School of Pharmaceutical Science São Paulo State University UNESP, Rodovia Araraquara/Jaú Km 01

Made available in DSpace on 2021-06-25T11:08:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-02-15 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Nanostructured polyelectrolyte complexes (nano PECs) based on biopolymers are an important technological strategy to target drugs to the action and/or absorption site in a more effective way. In this work, computational studies were performed to predict the ionization, spatial arrangement and interaction energies of chitosan (CS), hyaluronic acid (HA), and hypromellose phthalate (HP), for the design of nano PEC carriers for methotrexate (MTX). The optimal pH range (5.0–5.5) for preparing nano PECs was selected by experimental and computational methodologies, favoring the polymers interactions. CS, HA, HP and MTX addition order was also rationalized, maximizing their interactions and MTX entrapment. Spherical nano-sized particles (256–575 nm, by dynamic light scattering measurement) with positive surface charge (+25.5 to +29.2 mV) were successfully prepared. The MTX association efficiency ranged from 20 to 32 %. XRD analyses evidenced the formation of a new material with an organized structure, in relation to raw polymers. School of Pharmaceutical Sciences São Paulo State University (UNESP), Araraquara, Road Araraquara–Jaú, Km 01 Federal Institute of Mato Grosso (IFMT), Cuiabá Campus, Bela Vista, Juliano Costa Marques Avenue School of Pharmaceutical Sciences São Paulo State University (UNESP), Araraquara, Road Araraquara–Jaú, Km 01 FAPESP: 17/16324-0 FAPESP: 17/26349-0

Chitosan-based particles are widely proposed as biocompatible drug delivery systems with mucoadhesive and permeation enhancing properties. However, strategies on how to modulate the intended biological responses are still scarce. Considering that particle properties affect the biological outcome, the rational design of the synthesis variables should be proposed to engineer drug delivery systems with improved biological performance. The purpose of this review is to establish a deeper understanding of possible correlations between these variables and the particle properties from theoretical and experimental perspectives. The fundamental physicochemical knowledge of chitosan-based polyelectrolyte complexation and surface modification is discussed focusing on chitosan-TPP, polyelectrolyte complexes, and chitosan-surface modified PLGA or lipid particles. A set of design considerations is proposed to enable future investigation in the development of chitosan particles with modulated properties. The approach presented here contributes to the rational design of chitosan-based particles that meet different requirements for biological activities.

Made available in DSpace on 2020-12-12T02:13:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-08-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Cancer represents a significant public health problem. More than 18.1 million people are annually diagnosed with cancer and 9.6 million die mainly due to metastatic disease. Chemotherapy has been one of the main cancer treatment modalities; however, most of the chemotherapeutic agents are non-specific, exhibiting several toxic side effects, which compromises the patient's quality of life. Therefore, it is necessary to search for new therapeutic alternatives, using for example, drug delivery systems (DDS) to target cancer cells, increasing the selectivity of chemotherapeutic drugs. This approach is promising; however, it is crucial to evaluate the biological performance of the systems. Although mammalian models continue to be explored for clinical applications, they are time-consuming and very restrictive from the ethical and legal perspectives. Hence, the chick embryo chorioallantoic membrane (CAM) has been shown to be a suitable in vivo model since it allows a more appropriate model for the study of drugs and/or DDS performance than in vitro tests. Thereby, this article revises the recent advances of DDS for cancer therapy, evaluating the feasibility of the CAM model. UNESP São Paulo State University School of Pharmaceutical Sciences Department of Drugs and Medicines UNESP São Paulo State University School of Pharmaceutical Sciences Department of Clinical Analysis Life and Health Sciences Research Institute (ICVS) School of Medicine University of Minho ICVS/3B's-PT Government Associate Laboratory UNICAMP University of Campinas Piracicaba Dental School Department of Biosciences UNESP São Paulo State University School of Pharmaceutical Sciences Department of Drugs and Medicines UNESP São Paulo State University School of Pharmaceutical Sciences Department of Clinical Analysis FAPESP: #2013/01565-1 FAPESP: #2015/21412-0 FAPESP: #2016/06337-5, #2019/07245-5 FAPESP: #2016/09671-3 FAPESP: #2017/10016-2 FAPESP: #2017/23357-2 FAPESP: #2018/04546-1

This review discusses basic aspects used to control the architecture and functional properties of smart hydrogels. The introduction briefly outlines what has been accomplished regarding smart hydrogels and explores historical aspects and the fundamental understanding of these systems. Then, a short discussion on the chemical interactions and the main variables involved in architectural construction is exhibited. Further analysis provides the basis for optimizing biological responses through system modulation. Finally, we outline perspectives and challenges for building smart hydrogels into functionalized and modulated delivery systems.

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No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Submitted by Natália Noronha Ferreira Naddeo (natalia.noronha@unesp.br) on 2020-05-21T11:52:18Z No. of bitstreams: 2 TESE PARA REPOSITÓRIO_Parcial.pdf: 1859954 bytes, checksum: 286b1e73bd9ea5251a728a70f005cc75 (MD5) TESE PARA REPOSITÓRIO.pdf: 4670117 bytes, checksum: 25491ddc4b142eca2e79bfe7a8315346 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Submitted by Natália Noronha Ferreira Naddeo (natalia.noronha@unesp.br) on 2020-05-21T11:52:18Z No. of bitstreams: 2 TESE PARA REPOSITÓRIO_Parcial.pdf: 1859954 bytes, checksum: 286b1e73bd9ea5251a728a70f005cc75 (MD5) TESE PARA REPOSITÓRIO.pdf: 4670117 bytes, checksum: 25491ddc4b142eca2e79bfe7a8315346 (MD5) Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Submitted by Natália Noronha Ferreira Naddeo (natalia.noronha@unesp.br) on 2020-05-21T11:52:18Z No. of bitstreams: 2 TESE PARA REPOSITÓRIO_Parcial.pdf: 1859954 bytes, checksum: 286b1e73bd9ea5251a728a70f005cc75 (MD5) TESE PARA REPOSITÓRIO.pdf: 4670117 bytes, checksum: 25491ddc4b142eca2e79bfe7a8315346 (MD5) Submitted by Natália Noronha Ferreira Naddeo (natalia.noronha@unesp.br) on 2020-05-21T11:52:18Z No. of bitstreams: 2 TESE PARA REPOSITÓRIO_Parcial.pdf: 1859954 bytes, checksum: 286b1e73bd9ea5251a728a70f005cc75 (MD5) TESE PARA REPOSITÓRIO.pdf: 4670117 bytes, checksum: 25491ddc4b142eca2e79bfe7a8315346 (MD5) Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Submitted by Natália Noronha Ferreira Naddeo (natalia.noronha@unesp.br) on 2020-05-21T11:52:18Z No. of bitstreams: 2 TESE PARA REPOSITÓRIO_Parcial.pdf: 1859954 bytes, checksum: 286b1e73bd9ea5251a728a70f005cc75 (MD5) TESE PARA REPOSITÓRIO.pdf: 4670117 bytes, checksum: 25491ddc4b142eca2e79bfe7a8315346 (MD5) Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Submitted by Natália Noronha Ferreira Naddeo (natalia.noronha@unesp.br) on 2020-05-21T11:52:18Z No. of bitstreams: 2 TESE PARA REPOSITÓRIO_Parcial.pdf: 1859954 bytes, checksum: 286b1e73bd9ea5251a728a70f005cc75 (MD5) TESE PARA REPOSITÓRIO.pdf: 4670117 bytes, checksum: 25491ddc4b142eca2e79bfe7a8315346 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Submitted by Natália Noronha Ferreira Naddeo (natalia.noronha@unesp.br) on 2020-05-21T11:52:18Z No. of bitstreams: 2 TESE PARA REPOSITÓRIO_Parcial.pdf: 1859954 bytes, checksum: 286b1e73bd9ea5251a728a70f005cc75 (MD5) TESE PARA REPOSITÓRIO.pdf: 4670117 bytes, checksum: 25491ddc4b142eca2e79bfe7a8315346 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Submitted by Natália Noronha Ferreira Naddeo (natalia.noronha@unesp.br) on 2020-05-21T11:52:18Z No. of bitstreams: 2 TESE PARA REPOSITÓRIO_Parcial.pdf: 1859954 bytes, checksum: 286b1e73bd9ea5251a728a70f005cc75 (MD5) TESE PARA REPOSITÓRIO.pdf: 4670117 bytes, checksum: 25491ddc4b142eca2e79bfe7a8315346 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Submitted by Natália Noronha Ferreira Naddeo (natalia.noronha@unesp.br) on 2020-05-21T11:52:18Z No. of bitstreams: 2 TESE PARA REPOSITÓRIO_Parcial.pdf: 1859954 bytes, checksum: 286b1e73bd9ea5251a728a70f005cc75 (MD5) TESE PARA REPOSITÓRIO.pdf: 4670117 bytes, checksum: 25491ddc4b142eca2e79bfe7a8315346 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Submitted by Natália Noronha Ferreira Naddeo (natalia.noronha@unesp.br) on 2020-05-21T11:52:18Z No. of bitstreams: 2 TESE PARA REPOSITÓRIO_Parcial.pdf: 1859954 bytes, checksum: 286b1e73bd9ea5251a728a70f005cc75 (MD5) TESE PARA REPOSITÓRIO.pdf: 4670117 bytes, checksum: 25491ddc4b142eca2e79bfe7a8315346 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Submitted by Natália Noronha Ferreira Naddeo (natalia.noronha@unesp.br) on 2020-05-21T11:52:18Z No. of bitstreams: 2 TESE PARA REPOSITÓRIO_Parcial.pdf: 1859954 bytes, checksum: 286b1e73bd9ea5251a728a70f005cc75 (MD5) TESE PARA REPOSITÓRIO.pdf: 4670117 bytes, checksum: 25491ddc4b142eca2e79bfe7a8315346 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). 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No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-05-23T22:27:53Z (GMT) No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Made available in DSpace on 2020-05-23T22:27:53Z (GMT). No. of bitstreams: 1 colturato_pl_me_arafcf_par.pdf: 4011876 bytes, checksum: e36eae92e4d93289dafff9e35fb734b8 (MD5) Previous issue date: 2020-03-30 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Glioblastomas (GBM) representam 77% dos tumores malignos do sistema nervoso central (SNC) e ainda hoje, apesar de todos os avanços na terapia, continua com prognóstico limitado. A existência de barreiras fisiológicas como a barreira hematoencefálica (BHE) representa o principal obstáculo que impede que concentrações adequadas do fármaco atinjam o local de ação. Por suas vantagens anatômicas, uma estratégia proposta para a administração de fármacos destinados ao SNC consiste no uso da via nasal. Além disso, o uso de terapias combinadas utilizando fármacos capazes de agir em diferentes alvos moleculares deve ser considerada para o tratamento de doenças complexas como GBM. O candidato a fármaco ácido alfa-ciano-4-hidroxicinâmico (CHC) e o anticorpo monoclonal cetuximab (CTX) já são explorados devido à capacidade de agir em diferentes alvos moleculares nas células tumorais e aplicados em conjunto, como uma nova abordagem combinada, podem melhorar os resultados terapêuticos. De forma complementar, a utilização de sistemas de liberação baseados em nanotecnologia trará inevitavelmente ganhos terapêuticos à combinação proposta, permitindo que atributos específicos sejam agregados ao sistema e possibilite não somente a administração nasal, como também a associação de diferentes fármacos em um único carreador. Assim, o presente estudo propõe o desenvolvimento de diferentes plataformas poliméricas baseadas em poli(ácido láctico-co-glicólico) (PLGA) e quitosana trimetilada (TMC) ou quitosana oligomérica (OCS) para encapsulação do CHC. Ambos os sistemas desenvolvidos contendo CHC encapsulado exibiram tamanho de aproximadamente 300 a 400 nm contendo quitosana em sua superfície mais externa (potencial zeta positivo) e uma alta porcentagem de CHC encapsulado (± 85%). A caracterização físico-química dos sistemas mostrou sua estabilidade coloidal, principalmente na presença do fármaco CHC. Dados de DRX sugerem que a interação entre CHC e as nanopartículas (NPs) de PLGA/OCS apresentam padrões diferentes das NPs de PLGA/TMC. A conjugação entre CTX e NPs foi realizada através de ligações supramoleculares e covalente resultando em 85 e 58% de eficiência, respectivamente. A análise da eficácia terapêutica utilizando protocolos in vitro empregando linhas celulares de glioma U251 e SW1088, estabeleceu que, comparando ambos os sistemas conjugados, o PLGA/OCS parece ter uma capacidade terapêutica mais relevante. Por esse motivo, esse sistema foi selecionado para as futuras investigações. Análise de imunoblot confirmou que o CTX associado às NPs continua a exercer sua eficácia terapêutica. A análise da atividade antiangiogênica, desenvolvimento e progressão do tumor usando modelo de membrana corioalantóica de embrião de galinha (CAM) revelaram uma tendência de redução do tumor quando NPs conjugadas foram utilizadas. Além disso, elas também exibiram uma atividade antiangiogênica. O perfil de liberação in vitro mostrou que a liberação de CHC foi sustentada eretardada pelo seu encapsulamento nas NPs. Estudos de permeação ex vivo utilizando mucosa nasal suína mostraram que a permeação de CHC foi modificada pela sua inclusão nos sistemas. A avaliação da capacidade de NPs administradas pela via nasal em atingir o SNC utilizando tomografia de fluorescência forneceu evidências de independentemente do procedimento de conjugação, NPs foram eficazes em realizar o transporte direto para o cérebro pela rota nasal. Portanto, considerando todos os resultados mencionados, o sistema desenvolvido exibiu um conjunto de atributos favoráveis que as tornam alternativas promissoras a serem futuramente consideradas no tratamento de GBM. Glioblastomas (GBM) account for 77% of malignant tumors in the central nervous system (SNC), and today, despite all advances in therapy, remains with a limited prognosis. The existence of physiological barriers as the blood brain barrier (BBB) represents the main obstacle that limits appropriate concentrations of drugs designed to therapy. Due to their anatomical advantages, a strategy proposed for direct delivery to SNC involves the use of the nose-to-brain route. Besides, combination therapy that uses multiple drugs against different molecular targets should be considered for complex diseases such as GBM. Drugs like alpha-cyano-4-hydroxycinnamic acid (CHC) and the monoclonal antibody cetuximab (CTX) are already explored for their capacity to act against different hallmarks of cancer and applied together, as a novel combining approach, might improve therapeutic outcomes. Therefore, advances in nanotechnology-based delivery systems will inevitably bring therapeutic gains to the proposed combination since they enable acquisition of important characteristics desired and also the association of different drugs into a single carrier. Thus, the current study proposes the development of different polymeric platforms based on poly(lactic-co-glycolic acid) (PLGA) and trimethyl chitosan (TMC) /chitosan oligosaccharide (OCS) for CHC encapsulation. Both CHC-loaded developed systems (PLGA/TMC and PLGA/OCS) exhibited nanostructure organization of about 300 to 400 nm, containing chitosan on their outermost surface (positive zeta potential) and a high percentage of CHC encapsulation (±85%). Physicochemical characteristics have shown great colloidal stability, especially in the presence of CHC drug. DRX data suggest that interaction between CHC and PLGA/OCS NPs follows patterns different than PLGA/TMC NPs. Conjugation between CTX and developed CHCloaded NPs was optimally obtained by supramolecular forces and covalent bonds, resulting in 85 and 58% of efficacy, respectively. Analysis of therapeutic efficacy using in vitro protocols employing U251 and SW1088 glioma cell lines, established that, comparing both conjugated systems, PLGA/OCS seems to have greatly therapeutic capacity. Therefore, this system was chosen to further investigations. Blot analysis confirmed that CTX associated with NPs continues to exert its therapeutic efficacy. Analysis of antiangiogenic activity, tumor development, and progression using the chicken chorioallantoic membrane disclosed a trend of tumor reduction when conjugated NPs were employed. In addition, this system also exhibited antiangiogenic activity. In vitro release profile showed that CHC release was sustained and retarded by drug encapsulation into NPs. The ex vivo permeation study applying nasal porcine mucosal showed that CHC permeation was delayed by the inclusion of system complexity. Analysis of NPs delivery using fluorescence tomography provides evidence that the developed PLGA/OCS NPs, independently of conjugation procedure,were effective in providing nose-to-brain transport. Taking into consideration all of the aforementioned results, we anticipate that the developed system exhibited a set of favorable attributes that make them promising alternatives to be further considered in GBM treatment. FAPESP: 2016/09671-3

Made available in DSpace on 2020-12-12T02:34:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-02-01 Nanoparticles based on gellan gum/pectin blends were designed for colon-targeted release of resveratrol (RES). Their impact on drug release rates and permeability were evaluated using Caco-2 cell model and mucus secreting triple co-culture model. Polymeric nanoparticles (PNP) were successfully prepared by nebulization/ionotropic gelation, achieving high drug loading (>80%). PNP were spherical with a low positive charge density (+5mV) and exhibited diameters of around 330 nm. Developed PNP were able to promote effective modulation of drug release rates, so that only 3% of RES was released in acidic media over 2 h, and, in pH 6.8, the drug was released in a sustained manner, reaching 85% in 30 h. The permeability of RES-loaded PNP in the Caco-2 model was 0.15%, while in the triple co-culture model, in the presence of mucus, it reached 5.5%. The everted gut sac experiment corroborated the low permeability of RES-loaded PNP in the presence or absence of mucus and highlighted their high ability to interact with the intestinal tissue. Results indicate that the novel PNP developed in this work are safe and promising carriers for controlled delivery of RES at the colon. School of Pharmaceutical Sciences São Paulo State University (UNESP) Chemistry Institute of São Carlos São Paulo University (USP) I3S-Instituto de Investigação e Inovação em Saúde Universidade do Porto INEB-Instituto de Engenharia Biomédica Universidade do Porto CESPU-Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde School of Pharmaceutical Sciences São Paulo State University (UNESP)

Made available in DSpace on 2020-12-12T01:35:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-10-25 Fundação para a Ciência e a Tecnologia European Regional Development Fund Accumulating evidence has been suggesting that combining two or more anticancer drugs can provide additive or synergistic effects, improving therapeutic efficacy and delaying resistance. Nowadays, advances in nanotechnology-based delivery systems have enabled the association of different drugs into a single carrier and provided therapeutic gains to the proposed regimen. However, a new strategy also requires innovative analytical approaches that assess loading capacity, biological performance, and also comprehend the mechanisms of action. Alpha-cyano-4-hydroxycinnamic acid (CHC) and the monoclonal antibody (mAb) cetuximab (CTX) are explored worldwide for their therapeutic benefits against multiple cancer cells. The present work aims to develop and validate a new method for simultaneous quantification of CHC and CTX in nanoparticulate systems by using reverse phase high-performance liquid chromatography (RP-HPLC) with ultraviolet (UV) detection for CHC, and fluorescence detection for CTX. This method was designed following the guidelines of the International Conference on Harmonization ICH Q2 (R1) and the Food and Drug Administration (FDA) - Guidance for Bioanalytical Method Validation. Chromatographic separation was performed on a C18 column with the mobile phase composed by water, 0.1 % (v/v) trifluoroacetic acid (TFA) and acetonitrile (ACN)-0.1 % TFA on gradient mode at a flow rate of 0.6 mL/min. The performance of the present method was evaluated by system suitability; therefore, linearity, accuracy, precision, detection, limit of detection / limit of quantification, and robustness were also highlighted. Specificity was demonstrated by the chromatographic analyses of CHC and CTX, subjected to several informative stress conditions. The developed method was also successfully used for the first time to quantify the CHC and CTX content in poly(lactic-co-glycolic acid)-based nanoparticles. In conclusion, this new and rapid method presented acceptable analytical performance and can be helpful to simultaneously quantify CHC and CTX in future studies applied to anticancer therapy. School of Pharmaceutical Sciences São Paulo State University UNESP, Rodovia Araraquara-Jaú km 01, Araraquara Life and Health Sciences Research Institute (ICVS) School of Medicine University of Minho ICVS/3B's-PT Government Associate Laboratory School of Pharmaceutical Sciences São Paulo State University UNESP, Rodovia Araraquara-Jaú km 01, Araraquara

Made available in DSpace on 2018-12-11T16:52:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-06-01 Monoclonal antibody (mAb) delivery is gaining importance for local, systemic, and route-specific targeting. The mucus constitutes the main barrier for this type of delivery. In the present study, we aimed to develop a drug delivery platform by integrating mucus penetrating and mucoadhesive agents into a single system. Our hypothesis is that by combining these opposing functions, this system could have its properties modulated according to specific purposes. Self-assembly studies were conducted using three classes of building blocks: the protein drug bevacizumab (BVZ), mucus-penetrating polyanion dextran sulfate (DS), mucoadhesive polycations trimethylchitosan (TMC) and chitosan oligosaccharides (COS). We obtained two types of nanoparticles by manipulating supramolecular interactions between the components. Binary protein-polyanion (BVZ/DS) nanoparticles showed size of approximately 150 nm and a negative zeta potential. Ternary protein-polyanion-polycation (BVZ/DS/COS) nanoparticles were obtained using COS and exhibited 350 nm and a positive zeta potential. Assisted by calorimetric information, we demonstrated that building stable ternary nanoparticles carrying positive charges were not possible using the polycation TMC due to its thermodynamic constraints. Furthermore, spectroscopy analysis and CAM assay indicated that BVZ continued structurally and functionally stable after its incorporation into the nanoparticles. These two types of nanoparticles exhibited different behaviors when interacting with mucin, as shown by DLS and AFM studies. While the negatively charged particles promoted dispersion of the mucin network, suggesting a mucus penetrating effect of DS, the positively charged particles formed aggregates, probably caused by the mucoadhesive effect of COS. These results highlight the importance of understanding the role of supramolecular interactions, responsible for forming drug delivery systems containing complex molecules, such as proteins and polymers. School of Pharmaceutical Science São Paulo State University UNESP, Rodovia Araraquara/Jaú Km 1 Institute of Chemistry of Araraquara São Paulo State University UNESP Biophotonics Laboratory São Carlos Institute of Physics University of São Paulo (USP) EMBRAPA, Semi-Arid, Rodovia BE-428, Km 152, P.O. Box 23 EMBRAPA Instrumentation Rua XV de Novembro, 1.452, P.O. Box 741 Institute of Chemistry of São Carlos University of São Paulo USP School of Pharmaceutical Science São Paulo State University UNESP, Rodovia Araraquara/Jaú Km 1 Institute of Chemistry of Araraquara São Paulo State University UNESP

Made available in DSpace on 2018-12-11T17:35:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-01-08 European Regional Development Fund Fundação para a Ciência e a Tecnologia Ministério da Ciência, Tecnologia e Ensino Superior This work addresses the establishment and characterization of gellan gum:pectin (GG:P) biodegradable mucoadhesive beads intended for the colon-targeted delivery of resveratrol (RES). The impact of the polymer carrier system on the cytotoxicity and permeability of RES was evaluated. Beads of circular shape (circularity index of 0.81) with an average diameter of 914 μm, Span index of 0.29, and RES entrapment efficiency of 76% were developed. In vitro drug release demonstrated that beads were able to reduce release rates in gastric media and control release for up to 48 h at an intestinal pH of 6.8. Weibull's model correlated better with release data and b parameter (0.79) indicated that the release process was driven by a combination of Fickian diffusion and Case II transport, indicating that both diffusion and swelling/polymer chains relaxation are processes that contribute equally to control drug release rates. Beads and isolated polymers were observed to be safe for Caco-2 and HT29-MTX intestinal cell lines. RES encapsulation into the beads allowed for an expressive reduction of drug permeation in an in vitro triple intestinal model. This feature, associated with low RES release rates in acidic media, can favor targeted drug delivery from the beads in the colon, a promising behavior to improve the local activity of RES. Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista (UNESP), Araraquara, Rodovia Araraquara-Jaú, Km 1 Instituto de Química de São Carlos Universidade de São Paulo (USP), Avenida Trabalhador São-Carlense, 400 I3S-Instituto de Investigação e Inovação em Saúde Universidade do Porto, Rua Alfredo Allen, 208 INEB-Instituto de Engenharia Biomédica Universidade do Porto, Rua Alfredo Allen, 208 CESPU-Instituto de Investigacao e Formacao Avancada em Ciencias e Tecnologias da Saude, Rua Central de Gandra 1317 School of Pharmacy Queen's University Belfast Medical Biology Centre, 97 Lisburn Road Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista (UNESP), Araraquara, Rodovia Araraquara-Jaú, Km 1 Fundação para a Ciência e a Tecnologia: NORTE-01-0145-FEDER-000012 Fundação para a Ciência e a Tecnologia: SFRH/BD/118721/2016

Made available in DSpace on 2020-12-12T02:27:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-12-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) Hexagonal liquid crystals and supramolecular polymers from meglumine-based supra-amphiphiles were developed as drug delivery systems to be applied on the skin. The influence of fatty acid unsaturation on the structure and mechanical properties was evaluated. Moreover, we have investigated the system biocompatibility and how the type of water could influence its bioadhesive properties. Meglumine-oleic acid (MEG-OA) was arranged as hexagonal liquid crystals at 30–70 wt% water content, probably due to its curvature and increased water solubility. Meglumine-stearic acid (MEG-SA) at 10–80 wt% water content self-assembled as a lamellar polymeric network, which can be explained by the low mobility of MEG-SA in water due to hydrophobic interactions between fatty acid chains and H-bonds between meglumine and water molecules. Both systems have shown suitable mechanical parameters and biocompatibility, making them potential candidates to encapsulate therapeutic molecules for skin delivery. Moreover, a strong positive correlation between the amount of unfrozen bound water in meglumine-based systems and the bioadhesion properties was observed. This work shows that a better understanding of the physicochemical properties of a drug delivery system is extremely important for the correlation with the desired biological response and, thus, improve the product performance for biomedical applications. São Paulo State University (UNESP) Chemistry Institute, R. Prof. Francisco Degni, 55 São Paulo State University (UNESP) School of Pharmaceutical Sciences, Rodovia Araraquara˗Jau Km 1 Federal University of Rio de Janeiro (UFRJ) Department of Clinical and Toxicological Analysis Faculty of Pharmacy, Av. Carlos Chagas Filho 373 Federal University of Sergipe (UFS) Department of Chemistry, Campus Prof. Alberto de Carvalho, Av. Vereador Olímpio Grande s/n Department of Biology and Environment and Centre for Research and Technology of Agro-Environmental and Biological Sciences (CITAB) University of Trás-os-Montes e Alto Douro (UTAD), Quinta de Prados 1013 São Paulo State University (UNESP) Chemistry Institute, R. Prof. Francisco Degni, 55 São Paulo State University (UNESP) School of Pharmaceutical Sciences, Rodovia Araraquara˗Jau Km 1 FAPESP: #2013/08411-0

Alginate hydrogel improves anti-angiogenic bevacizumab activity in cancer therapy, Anti-vascular endothelial growth factor (anti-VEGF) therapy applied to solid tumors is a promising strategy, yet, the challenge to deliver these agents at high drug concentrations together with the maintenance of therapeutic doses locally, at the tumor site, minimizes its benefits. To overcome these obstacles, we propose the development of a bevacizumab-loaded alginate hydrogel by electrostatic interactions to design a delivery system for controlled and anti-angiogenic therapy under tumor microenvironrnental conditions. The tridimensional hydrogel structure produced provides drug stability and a system able to be introduced as a flowable solution, stablishing a depot after local administration. Biological performance by the chick embryo chorioallantoic membrane (CAM) assay indicated a pH-independent improved anti-angiogenic activity (similar to 50%) compared to commercial available anti-VEGF drug. Moreover, there was a considerable regression in tumor size when treated with this system. Immunohistochemistry highlighted a reduced number and disorganization of microscopic blood vessels resulting from applied therapy. These results suggest that the developed hydrogel is a promising approach to create an innovative delivery system.that offers the possibility to treat different solid tumors by intratumoral administration., Brazilian Fundação de Amparo e Pesquisa do Estado de São Paulo (FAPESP), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). Additionally, this article has been developed under the scope of the project NORTE-01-0145-FEDER-000013, supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER) Project PTDC/SAU-TOX/114549/2009 – FCOMP-01-0124-FEDER-016057, through the Competitiveness Factors Operational Programme (COMPETE), and by National funds, through the Foundation for Science and Technology (FCT), under the scope of the project POCI-01-0145-FEDER-007038, info:eu-repo/semantics/publishedVersion

Made available in DSpace on 2018-12-11T17:32:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-10-03 This work aimed to develop a calcium alginate hydrogel as a pH responsive delivery system for polymyxin B (PMX) sustained-release through the vaginal route. Two samples of sodium alginate from different suppliers were characterized. The molecular weight and M/G ratio determined were, approximately, 107 KDa and 1.93 for alginate_S and 32 KDa and 1.36 for alginate_V. Polymer rheological investigations were further performed through the preparation of hydrogels. Alginate_V was selected for subsequent incorporation of PMX due to the acquisition of pseudoplastic viscous system able to acquiring a differential structure in simulated vaginal microenvironment (pH 4.5). The PMX-loaded hydrogel (hydrogel_PMX) was engineered based on polyelectrolyte complexes (PECs) formation between alginate and PMX followed by crosslinking with calcium chloride. This system exhibited a morphology with variable pore sizes, ranging from 100 to 200 μm and adequate syringeability. The hydrogel liquid uptake ability in an acid environment was minimized by the previous PECs formation. In vitro tests evidenced the hydrogels mucoadhesiveness. PMX release was pH-dependent and the system was able to sustain the release up to 6 days. A burst release was observed at pH 7.4 and drug release was driven by an anomalous transport, as determined by the Korsmeyer–Peppas model. At pH 4.5, drug release correlated with Weibull model and drug transport was driven by Fickian diffusion. The calcium alginate hydrogels engineered by the previous formation of PECs showed to be a promising platform for sustained release of cationic drugs through vaginal administration. School of Pharmaceutical Sciences São Paulo State University UNESP Department of Chemistry Federal University of São Carlos School of Pharmaceutical Sciences São Paulo State University UNESP

Humic substances (HS) are heterogeneous molecular aggregates, which have no defined structure and constitute the principal fraction of natural organic matter. They can act as complexation agents, due to their high density of functional groups, and therefore directly influence the transport, accumulation, bioavailability and toxicity of different substances. Metals are important environmental pollutants, since they are widely used during human activities, and following release are not either chemically or biologically degraded. Treatment of acute poisoning in humans caused by potentially toxic metals has employed complexation agents such as ethylenediaminetetraacetic acid (EDTA) or -aminoacids. However, some limitations of these materials such as the lack of selectivity of EDTA and the high cost of -aminoacids have provided the incentive for a search for alternatives. Previous studies of the properties of HS indicate their possible use as an alternative to these chelanting agents. Substâncias húmicas (SH) são agregados moleculares heterogêneos, possuem estrutura indefinida e constituem a principal fração da matéria orgânica natural. Devido ao elevado número de grupos funcionais presentes em sua estrutura, podem atuar como agentes complexantes, influenciando diretamente o transporte, acúmulo, biodisponibilidade e toxicidade de diferentes substâncias. Dentre os contaminantes ambientais, se destacam os metais, pela sua ampla aplicabilidade em diferentes setores e por não sofrerem degradação química ou biológica. O tratamento de intoxicações por metais potencialmente tóxicos em seres humanos vem sendo realizado por agentes complexantes, como o etilenodiaminotetraácetico (EDTA) e os -aminoácidos. Entretanto, algumas limitações, como a falta de seletividade do EDTA e o elevado custo dos -aminoácidos, têm incentivado a busca por outras substâncias quelantes. Nesse sentido, este artigo aborda as principais propriedades descritas na literatura que tornam as substâncias húmicas uma interessante alternativa como agente terapêutico, em casos de intoxicações por metais potencialmente tóxicos.

Resumo: A iodoterapia é uma das terapêuticas da medicina nuclear que emprega como radionuclídeo o iodo radioativo. Esse tratamento, realizado em unidades de internação, exige procedimentos de radioproteção para prevenir contaminação de pacientes, equipe de saúde e meio ambiente. Uma discussão sobre a aplicação dessa terapêutica mostra a importância da implantação de programas de gerenciamento de resíduos radioativos, gerados em unidades hospitalares.