Antinociceptive and in vitro anticancer activity of microencapsulated extracts and vouacapans isolated from Pterodon pubescens Benth

Orientadores: Mary Ann Foglio, João Ernesto de Carvalho, Rodney Alexandre Ferreira, RodriguesDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: As plantas medicinais têm sido fonte de agentes medicinais há milhares de anos para o tratamento de um amplo espectro de doenças. Entre as alternativas que podem ser utilizadas para preservar as características do material vegetal, destaca-se a encapsulação em micropartículas. Estudos prévios com espécie Pterodon pubescens Benth. (sucupira) avaliaram a eficácia (ação antiinflamatória e antinociceptiva) e a segurança (eventuais efeitos toxicológicos) dos extratos, frações ativas e compostos isolados de P. pubescens. No presente trabalho, avaliou-se o benefício do processo de microencapsulação por spray drying para preservação das características químicas, aliado a ações biológicas in vitro e in vivo dos extratos e compostos isolados. Extratos brutos produzidos com sementes provenientes de Minas Gerais (EBMG), Bahia (EBBA), Mato Grosso (EBMT) ou São Paulo (EBSP) apresentaram perfil químico distinto, demonstrando a variabilidade química da espécie de diferentes procedências. A partir dos extratos EBMG e EBSP foram isolados os vouacapanos C1 e C2, ambos com atividade antinociceptiva, sendo que a atividade de C1 esteve mais relacionada a componente de dor neurogênica. C1 e os extratos EBBA, EBSP e EBMT foram microencapsulados por spray drying e, juntamente com as amostras livres, armazenados em câmara climática com umidade e temperatura controladas para o estudo de estabilidade. Os resultados dos ensaios de atividade anticâncer in vitro em células tumorais humanas e atividade antinociceptiva em modelos com camundongos, demonstraram que a técnica de microencapsulação por spray drying preservou a ação, tanto do composto isolado quanto dos extratos demonstrando ser uma alternativa útil para prolongar o tempo de estocagem, principalmente do extrato bruto.As vantagens apresentadas pelo processo de microencapsulação dos produtos obtidos a partir de P. pubescens, motivam a continuidade dos estudos de padronização com esta espécie, para viabilizar um produto que atenda a parâmetros de reprodutibilidade, eficácia e segurança.Abstract: Over the years natural products, especially plants, have demonstrated to play a dominant role for drug discovery to treat human diseases. Micro encapsulation is an important approach to preserve natural products characteristics. Previous studies with Pterodon pubescens Benth., (sucupira) evaluated the efficacy (anti-inflammatory and antinociceptive) and safety (possible toxicological effects) of the extracts, active fractions and compounds isolated from P. pubescens. In the present study we evaluated the benefit of the microencapsulation process by spray drying to preserve the chemical characteristic monitored by in vitro and in vivo biological activities. Crude extracts obtained from Minas Gerais (EBMG), Bahia (EBBA), Mato Grosso (EBMT) or São Paulo (EBSP) seeds showed different quimical profiles. Vouacapans C1 and C2, isolated to EBMG and EBSP crude extracts, demonstrated antinociceptive activity, C1 activity related to neurogenic pain. The crude extracts EBBA, EBSP, EBMT and compound C1 were microencapsulated by spray drying and, along with control samples, stored in a climatic chamber with controlled humidity and temperature for the accelerated stability studies. The test results of in vitro anticancer activity in human tumor cells and antinociceptive activity in mice models, showed that the microencapsulation technique by spray drying preserve both the isolated compound and extracts. This technique is a useful alternative to extend storage time, especially of the crude extract. The advantages presented by P. pubescens microencapsulated products prompt to continue studies with this species, for the production of a product that meets reproductively, efficacy and safety parameters.MestradoFarmacologia, Anestesiologia e TerapeuticaMestre em Odontologi

Chitosan-tripolyphosphate Nanoparticles As Arrabidaea Chica Standardized Extract Carrier: Synthesis, Characterization, Biocompatibility, And Antiulcerogenic Activity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Natural products using plants have received considerable attention because of their potential to treat various diseases. Arrabidaea chica (Humb. & Bonpl.) B. Verlot is a native tropical American vine with healing properties employed in folk medicine for wound healing, inflammation, and gastrointestinal colic. Applying nanotechnology to plant extracts has revealed an advantageous strategy for herbal drugs considering the numerous features that nanostructured systems offer, including solubility, bioavailability, and pharmacological activity enhancement. The present study reports the preparation and characterization of chitosan-sodium tripolyphosphate nanoparticles (NPs) charged with A. chica standardized extract (AcE). Particle size and zeta potential were measured using a Zetasizer Nano ZS. The NP morphological characteristics were observed using scanning electron microscopy. Our studies indicated that the chitosan/sodium tripolyphosphate mass ratio of 5 and volume ratio of 10 were found to be the best condition to achieve the lowest NP sizes, with an average hydrodynamic diameter of 150 +/- 13 nm and a zeta potential of +45 +/- 2 mV. Particle size decreased with AcE addition (60 +/- 10.2 nm), suggesting an interaction between the extract's composition and polymers. The NP biocompatibility was evaluated using human skin fibroblasts. AcE-NP demonstrated capability of maintaining cell viability at the lowest concentrations tested, stimulating cell proliferation at higher concentrations. Antiulcerogenic activity of AcE-NP was also evaluated with an acute gastric ulcer experimental model induced by ethanol and indomethacin. NPs loaded with A. chica extract reduced the ulcerative lesion index using lower doses compared with the free extract, suggesting that extract encapsulation in chitosan NPs allowed for a dose reduction for a gastroprotective effect. The AcE encapsulation offers an approach for further application of the A. chica extract that could be considered a potential candidate for ulcer-healing pharmaceutical systems.1038973909Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)FAPESP [2009/16484-1, 2010/07781-0, 2009/51020-6, 2011/09127-8

Chitosan-tripolyphosphate Nanoparticles As Arrabidaea Chica Standardized Extract Carrier: Synthesis, Characterization, Biocompatibility, And Antiulcerogenic Activity.

Natural products using plants have received considerable attention because of their potential to treat various diseases. Arrabidaea chica (Humb. & Bonpl.) B. Verlot is a native tropical American vine with healing properties employed in folk medicine for wound healing, inflammation, and gastrointestinal colic. Applying nanotechnology to plant extracts has revealed an advantageous strategy for herbal drugs considering the numerous features that nanostructured systems offer, including solubility, bioavailability, and pharmacological activity enhancement. The present study reports the preparation and characterization of chitosan-sodium tripolyphosphate nanoparticles (NPs) charged with A. chica standardized extract (AcE). Particle size and zeta potential were measured using a Zetasizer Nano ZS. The NP morphological characteristics were observed using scanning electron microscopy. Our studies indicated that the chitosan/sodium tripolyphosphate mass ratio of 5 and volume ratio of 10 were found to be the best condition to achieve the lowest NP sizes, with an average hydrodynamic diameter of 150±13 nm and a zeta potential of +45±2 mV. Particle size decreased with AcE addition (60±10.2 nm), suggesting an interaction between the extract's composition and polymers. The NP biocompatibility was evaluated using human skin fibroblasts. AcE-NP demonstrated capability of maintaining cell viability at the lowest concentrations tested, stimulating cell proliferation at higher concentrations. Antiulcerogenic activity of AcE-NP was also evaluated with an acute gastric ulcer experimental model induced by ethanol and indomethacin. NPs loaded with A. chica extract reduced the ulcerative lesion index using lower doses compared with the free extract, suggesting that extract encapsulation in chitosan NPs allowed for a dose reduction for a gastroprotective effect. The AcE encapsulation offers an approach for further application of the A. chica extract that could be considered a potential candidate for ulcer-healing pharmaceutical systems.103897-390

Arrabidaea chica Verlot : controled release formulations with aplications for ulcer lesions

Orientadores: Mary Ann Foglio, João Ernesto de Carvalho, Rodney Alexandre Ferreira RodriguesTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: A espécie Arrabidaea chica, conhecida como crajiru, faz parte da relação nacional de plantas medicinais de interesse ao SUS. Estudos desenvolvidos no CPQBA-UNICAMP comprovaram que o extrato bruto de A. chica possui ação cicatrizante, antiulcerogênica e antioxidante. Este trabalho teve como objetivo delinear e avaliar sistemas farmacêuticos contendo o extrato padronizado de A. chica, para o tratamento de lesões de pele e mucosa. Foram obtidas nanopartículas de quitosana pelo método de reticulação. Dentre os parâmetros avaliados estipulou-se como melhores condições para produção de nanopartículas o emprego do tripolifosfato como reticulante nas proporções de 1:5 (massa) e 1:10 (volume) em relação a quitosana. Para as aplicações tópicas, as nanopartículas de A. chica foram veiculadas em esponjas, filmes absorvíveis de álcool polivinílico ou hidrogéis de ácido hialurônico. Estudos in vitro utilizando fibroblastos humanos demonstraram a atividade das nanopartículas de A. chica na proliferação e migração celular. Estudos in vivo empregando ratos e hamsters forneceram parâmetros para padronização de modelos de mucosite gastrointestinal e oral, respectivamente. A atividade das nanopartículas em úlceras de mucosa foi evidenciada nos modelos de úlcera gástrica induzidas por etanol ou indometacina, com redução das lesões ulcerativas de 76% e 58%, respectivamente, quando comparadas ao grupo controle negativo. O efeito cicatrizante de filmes absorvíveis ou hidrogéis incorporados com as nanopartículas de A. chica foram avaliados em modelo de úlcera dérmica em ratos. A contração da área da ferida chegou a 79% nos animais tratados com os filmes carregados com 0,5 mg de nanopartículas e 85% de contração nos animais que receberam hidrogéis contendo 1,5 mg de nanopartículas. Os resultados demonstraram que a produção de nanopartículas de A. chica e dos sistemas de transporte foi viável, caracterizando-se como uma alternativa válida para a veiculação do extrato de A. chica, além de propiciar a redução da dose necessária para a atividadeAbstract: Arrabidaea chica, known as crajiru, is part of the national Unified Health System from Brazil (SUS) list of medicinal plants of interest. Studies conducted at CPQBA- UNICAMP showed that the species demonstrated wound healing, antiulcerogenic and antioxidant properties. This study aimed the evaluation of pharmaceutical systems employing standardized extract for treatment of skin and mucosal wounds. Chitosan nanoparticles were obtained using the crosslink method. The protocol that showed best conditions to produce nanoparticles, among those tested, used tripolyphosphate as a cross-linker, in 1:5 (mass) and 1:10 (volume) proportions compared to chitosan. For topical applications, A. chica nanoparticles were incorporated into sponges, polyvinyl alcohol absorbable films or hyaluronic acid hydrogels. In vitro studies with human fibroblasts showed the activity of A. chica nanoparticles in cell proliferation and migration. In vivo studies with rats and hamsters provided the standardization parameters for gastrointestinal and mucositis models respectively. A chica nanoparticles demonstrated inhibition of mucosal ulcerations of gastric ulcer induced by ethanol or indomethacin models, improving ulcerative wound healing by 76% and by 58% respectively, in comparison to negative control groups. Those same samples incorporated into absorbable films or hydrogels were evaluated on wound healing activity using a rat dermal ulcer model. Animals treated with loaded films with 0.5 mg of nanoparticles achieved 79% in wound area contraction, whereas animals receiving hydrogels containing 1.5 mg of nanoparticles achieved 85%. These results showed that the production of A. chica nanoparticles and transport systems were practicable, demonstrating to be a valid alternative for incorporation of A. chica extracts, allowing decrease of the required dose to achieve the desired effectDoutoradoFarmacologia, Anestesiologia e TerapeuticaDoutora em Odontologi

Chitosan-tripolyphosphate Nanoparticles As Arrabidaea Chica Standardized Extract Carrier: Synthesis, Characterization, Biocompatibility, And Antiulcerogenic Activity.

Natural products using plants have received considerable attention because of their potential to treat various diseases. Arrabidaea chica (Humb. & Bonpl.) B. Verlot is a native tropical American vine with healing properties employed in folk medicine for wound healing, inflammation, and gastrointestinal colic. Applying nanotechnology to plant extracts has revealed an advantageous strategy for herbal drugs considering the numerous features that nanostructured systems offer, including solubility, bioavailability, and pharmacological activity enhancement. The present study reports the preparation and characterization of chitosan-sodium tripolyphosphate nanoparticles (NPs) charged with A. chica standardized extract (AcE). Particle size and zeta potential were measured using a Zetasizer Nano ZS. The NP morphological characteristics were observed using scanning electron microscopy. Our studies indicated that the chitosan/sodium tripolyphosphate mass ratio of 5 and volume ratio of 10 were found to be the best condition to achieve the lowest NP sizes, with an average hydrodynamic diameter of 150±13 nm and a zeta potential of +45±2 mV. Particle size decreased with AcE addition (60±10.2 nm), suggesting an interaction between the extract's composition and polymers. The NP biocompatibility was evaluated using human skin fibroblasts. AcE-NP demonstrated capability of maintaining cell viability at the lowest concentrations tested, stimulating cell proliferation at higher concentrations. Antiulcerogenic activity of AcE-NP was also evaluated with an acute gastric ulcer experimental model induced by ethanol and indomethacin. NPs loaded with A. chica extract reduced the ulcerative lesion index using lower doses compared with the free extract, suggesting that extract encapsulation in chitosan NPs allowed for a dose reduction for a gastroprotective effect. The AcE encapsulation offers an approach for further application of the A. chica extract that could be considered a potential candidate for ulcer-healing pharmaceutical systems

Chitosan-tripolyphosphate nanoparticles as arrabidaea chica standardized extract carrier: synthesis, characterization, biocompatibility, and antiulcerogenic activity

Natural products using plants have received considerable attention because of their potential to treat various diseases. Arrabidaea chica (Humb. & Bonpl.) B. Verlot is a native tropical American vine with healing properties employed in folk medicine for wound healing, inflammation, and gastrointestinal colic. Applying nanotechnology to plant extracts has revealed an advantageous strategy for herbal drugs considering the numerous features that nanostructured systems offer, including solubility, bioavailability, and pharmacological activity enhancement. The present study reports the preparation and characterization of chitosan-sodium tripolyphosphate nanoparticles (NPs) charged with A. chica standardized extract (AcE). Particle size and zeta potential were measured using a Zetasizer Nano ZS. The NP morphological characteristics were observed using scanning electron microscopy. Our studies indicated that the chitosan/sodium tripolyphosphate mass ratio of 5 and volume ratio of 10 were found to be the best condition to achieve the lowest NP sizes, with an average hydrodynamic diameter of 150 +/- 13 nm and a zeta potential of +45 +/- 2 mV. Particle size decreased with AcE addition (60 +/- 10.2 nm), suggesting an interaction between the extract's composition and polymers. The NP biocompatibility was evaluated using human skin fibroblasts. AcE-NP demonstrated capability of maintaining cell viability at the lowest concentrations tested, stimulating cell proliferation at higher concentrations. Antiulcerogenic activity of AcE-NP was also evaluated with an acute gastric ulcer experimental model induced by ethanol and indomethacin. NPs loaded with A. chica extract reduced the ulcerative lesion index using lower doses compared with the free extract, suggesting that extract encapsulation in chitosan NPs allowed for a dose reduction for a gastroprotective effect. The AcE encapsulation offers an approach for further application of the A. chica extract that could be considered a potential candidate for ulcer-healing pharmaceutical systems1038973909CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informação2009/16484-1; 2010/07781-0; 2009/51020-6; 2011/09127-