Assuntos
Antineoplásicos , Complexos de Coordenação , Neoplasias , Elementos de Transição , Antineoplásicos/química , Antineoplásicos/farmacologia , Humanos , Complexos de Coordenação/química , Complexos de Coordenação/farmacologia , Elementos de Transição/química , Neoplasias/tratamento farmacológicoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: In Brazilian popular medicine, Lippia alba leaves are used in teas to treat pain and inflammatory diseases. AIM OF THE STUDY: to evaluate the chemical composition, antinociceptive, and anti-inflammatory activities of Lippia alba essential oil and its major compound geraniol. MATERIAL AND METHODS: Lippia alba leaves were collected in Pará state, Brazil. The leaf essential oil was obtained using a modified Clevenger-type extractor. Then, the oil was analyzed by GC and GC-MS analyses. To evaluate the toxicity of LaEO and geraniol, the doses of 50, 300, and 2000 mg/kg were used in a mouse model. For antinociception tests, abdominal contortion, hot plate, and formalin tests were used; all groups were treated with LaEO and geraniol at doses of 25, 50, and 100 mg/kg; and to evaluate inflammation using the ear edema model. RESULTS: The constituents identified in the highest content were oxygenated monoterpenes: geraniol (37.5%), geranial (6.7%) and neral (3.8%). The animals treated with LaEO and geraniol demonstrated atypical behaviors with aspects of lethargy and drowsiness, characteristics of animals in a state of sedation; the relative weights showed no significant difference compared to the controls. In the abdominal contortion test, LaEO at 25 mg/kg, 50 mg/kg doses, and 100 mg/kg reduced the number of contortions, representing a percentage reduction of 84.64%, 81.23%, and 66.21% respectively. In the hot plate test, LaEO and geraniol increased the latency time at doses of 25, 50, and 100 mg/kg in all test periods; there was no statistical difference between LaEO and geraniol. In the first phase of the formalin test, only doses of 25 mg/kg and 100 mg/kg of LaEO showed significant activity, reducing the latency time by 53.40% and 58.90%. LaEO at doses of 25 mg/kg and 100 mg/kg reduced the size of the edema, demonstrating an anti-inflammatory activity of 59.38% (25 mg/kg) and 50% (100 mg/kg). CONCLUSION: Lippia alba essential oil and geraniol showed central/peripheral analgesic and anti-inflammatory potential and can be used as an alternative or complementary treatment to conventional drugs. More studies are needed to evaluate its action mechanisms and its analgesic effects.
Assuntos
Monoterpenos Acíclicos , Analgésicos , Anti-Inflamatórios , Edema , Lippia , Óleos Voláteis , Folhas de Planta , Animais , Lippia/química , Óleos Voláteis/farmacologia , Óleos Voláteis/química , Brasil , Analgésicos/farmacologia , Analgésicos/isolamento & purificação , Camundongos , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/isolamento & purificação , Masculino , Folhas de Planta/química , Edema/tratamento farmacológico , Edema/induzido quimicamente , Monoterpenos Acíclicos/farmacologia , Plantas Medicinais/química , Dor/tratamento farmacológico , Comportamento Animal/efeitos dos fármacos , Relação Dose-Resposta a Droga , Medição da Dor/efeitos dos fármacosRESUMO
Cellular senescence is recognized as a dynamic process in which cells evolve and adapt in a context dependent manner; consequently, senescent cells can exert both beneficial and deleterious effects on their surroundings. Specifically, senescent mesenchymal stromal cells (MSC) in the bone marrow (BM) have been linked to the generation of a supporting microenvironment that enhances malignant cell survival. However, the study of MSC's senescence role in leukemia development has been straitened not only by the availability of suitable models that faithfully reflect the structural complexity and biological diversity of the events triggered in the BM, but also by the lack of a universal, standardized method to measure senescence. Despite these constraints, two- and three dimensional in vitro models have been continuously improved in terms of cell culture techniques, support materials and analysis methods; in addition, research on animal models tends to focus on the development of techniques that allow tracking leukemic and senescent cells in the living organism, as well as to modify the available mice strains to generate individuals that mimic human BM characteristics. Here, we present the main advances in leukemic niche modeling, discussing advantages and limitations of the different systems, focusing on the contribution of senescent MSC to leukemia progression.
Assuntos
Leucemia , Células-Tronco Mesenquimais , Animais , Medula Óssea/patologia , Senescência Celular , Leucemia/patologia , Camundongos , Microambiente TumoralRESUMO
Antioxidants have drawn the attention of the scientific community due to being related to the prevention of various degenerative diseases. The antioxidant capacity has been extensively studied in vitro, and different methods have been used to assess its activity. However, the main issues related to studying natural antioxidants are evaluating whether these antioxidants demonstrate a key role in the biological system and assessing their bioavailability in the organism. The majority of outcomes in the literature are controversial due to a lack of method standardization and their proper application. Therefore, this study aims to compile the main issues concerning the natural antioxidant field of study, comparing the most common in vitro methods to evaluate the antioxidant activity of natural compounds, demonstrating the antioxidant activity in biological systems and the role of the main antioxidant enzymes of redox cellular signaling and explaining how the bioavailability of bioactive compounds is evaluated in animal models and human clinical trials.
Assuntos
Antioxidantes , Animais , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Disponibilidade Biológica , OxirreduçãoRESUMO
The CRISPR/Cas9 system enables a versatile set of genomes editing and genetic-based disease modeling tools due to its high specificity, efficiency, and accessible design and implementation. In cancer, the CRISPR/Cas9 system has been used to characterize genes and explore different mechanisms implicated in tumorigenesis. Different experimental strategies have been proposed in recent years, showing dependency on various intrinsic factors such as cancer type, gene function, mutation type, and technical approaches such as cell line, Cas9 expression, and transfection options. However, the successful methodological approaches, genes, and other experimental factors have not been analyzed. We, therefore, initially considered more than 1,300 research articles related to CRISPR/Cas9 in cancer to finally examine more than 400 full-text research publications. We summarize findings regarding target genes, RNA guide designs, cloning, Cas9 delivery systems, cell enrichment, and experimental validations. This analysis provides valuable information and guidance for future cancer gene validation experiments.
Assuntos
Sistemas CRISPR-Cas , Neoplasias , Edição de Genes , Humanos , Mutação , Neoplasias/genética , Oncogenes , RNA Guia de Cinetoplastídeos/genéticaRESUMO
Salivary gland carcinomas (SGC) are aggressive cancers that arise in minor and major salivary glands. Given the complexity and the multiple subtypes of this class of tumors, diagnosis and, treatment may be challenging for clinicians. Recently the tumor microenvironment, composed mainly of immune and stromal cells are been a target for treatment. Accumulating evidence indicates that cancer immunotherapies have made a significant impact on oncologic patients, however immunotherapeutic attempts in SGC have been shown limited improvement. Advances in the models that best translate aggressive SGC are needed for the development of clinical protocols grouping immunotherapies and other classes of drugs that will promote better responses in patients with advanced SGC stages. In this review, we introduced different experimental models for SGC with a focus on tumor microenvironment highlighting potential therapy applications for each model.
Assuntos
Carcinoma , Neoplasias das Glândulas Salivares , Humanos , Imunoterapia , Neoplasias das Glândulas Salivares/patologia , Neoplasias das Glândulas Salivares/terapia , Glândulas Salivares/patologia , Microambiente TumoralRESUMO
En la actualidad, la investigación biomédica se ha centrado en el estudio de enfermedades como el cáncer, que causan un elevado índice de mortalidad. Existen diferentes modelos animales, empleados para generar diversos tipos de carcinogénesis; el daño directo al ADN es uno de los mecanismos más utilizados. Sin embargo, en la normatividad nacional e internacional vigente, no se señalan los aspectos bioéticos que se deben seguir para desarrollar un modelo experimental de daño al ADN. Además, no se realiza una correcta semejanza de la enfermedad. Debido a lo anterior, esta revisión analiza los avances en cuanto a normatividad que se han generado en diferentes países, comparando los estudios encontrados en Estados Unidos, México y España. La perspectiva a futuro es poder contar con guías de experimentación actualizadas, que permitan pautar las normas necesarias para el adecuado desarrollo de los modelos de investigación animal de daño al ADN y que cumplan con la regla de las 3R en la experimentación animal. Esta iniciativa se debe de realizar en conjunto entre la Organización Mundial de la Salud y los organismos especializados en manejo y cuidado de animales de laboratorio en los ámbitos nacional e internacional.
Currently, biomedical research has focused on the study of diseases such as cancer that causes a high mortality rate. Different animal models are used to generate different types of carcinogenesis; direct DNA damage is one of the most used mechanisms. However, current national and international regulations do not indicate the bioethical aspects that must be followed to develop an experimental model of DNA damage. In addition, they do not perform a correct resemblance of the disease. Due to the above, this review analyzes the advances in regulations generated in different countries, comparing the studies found in the United States, Mexico, and Spain. The future perspective is to be able to count on updated experimentation guidelines, which allow the establishment of the necessary norms for the adequate development of animal research models of DNA damage that comply with the 3R rule in animal experimentation. This initiative should be carried out jointly by the World Health Organization and organizations specialized in managing and caring laboratory animals at the national and international levels.
Na atualidade, a pesquisa biomédica vem se focando no estudo de doenças que causam um elevado índice de mortalidade, como o câncer. Existem diferentes modelos animais utilizados para gerar diversos tipos de carcinogêneses; o dano direto ao DNA é um dos mecanismos mais utilizados. Contudo, na legislação nacional e internacional vigente, não são sinalizados os aspectos bioéticos que devem ser seguidos para desenvolver um modelo experimental de dano ao DNA, além de não ser realizada uma correta semelhança da doença. Devido a isso, esta revisão analisa o avanço quanto à legislação que vem sendo gerada em diferentes países, comparando os estudos encontrados nos Estados Unidos, no México e na Espanha. A perspectiva para o futuro é poder contar com atualizadas, que permitam estabelecer as normas necessárias para desenvolver os modelos de pesquisa animal de dano ao DNA e que cumpram com a regra das 3R na experimentação animal. Essa iniciativa se deve realizar em conjunto entre a Organização Mundial da Saúde e as organizações especializadas na gestão e cuidado de animais de laboratório nos contextos nacional e internacional.
RESUMO
The emergence and rapid worldwide spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has prompted the scientific community to rapidly develop in vitro and in vivo models that could be applied in COVID-19 research. In vitro models include two-dimensional (2D) cultures of immortalized cell lines or primary cells and three-dimensional (3D) cultures derived from lung, alveoli, bronchi, and other organs. Although cell-based systems are economic and allow strict control of experimental variables, they do not always resemble physiological conditions. Thus, several in vivo models are being developed, including different strains of mice, hamsters, ferrets, dogs, cats, and non-human primates. In this review, we summarize the main models of SARS-CoV-2 infection developed so far and discuss their advantages, drawbacks and main uses.
Assuntos
COVID-19/virologia , Modelos Animais de Doenças , Técnicas In Vitro/métodos , SARS-CoV-2/fisiologia , Animais , Linhagem Celular , Humanos , Pandemias , SARS-CoV-2/patogenicidade , Replicação ViralRESUMO
In the last two decades, alginate scaffolds have been variously studied as extracellular matrix analogs for tissue engineering. However, relevant evidence is still lacking concerning their ability to mimic the microenvironment of hierarchical tissues such as bone. Hence, an increasing amount of attention has recently been devoted to the fabrication of macro/microporous sponges with pore anisotropy able to more accurately replicate the cell niche structure as a trigger for bioactive functionalities. This paper presents an in vivo study of alginate sponges with anisotropic microporous domains (MAS) formed by ionic crosslinking in the presence of different fractions (30 or 50% v) of hydroxyapatite (HA). In comparison with unloaded sponges (MAS0), we demonstrated that HA confers peculiar physical and biological properties to the sponge, depending upon the inorganic fraction used, enabling the sponge to bio-mimetically support the regeneration of newly formed bone. Scanning electron microscopy analysis showed a preferential orientation of pores, ascribable to the physical constraints exerted by HA particles during the pore network formation. Energy dispersive spectroscopy (EDS) and X-Ray diffraction (XRD) confirmed a chemical affinity of HA with the native mineral phase of the bone. In vitro studies via WST-1 assay showed good adhesion and proliferation of human Dental Pulp-Mesenchymal Stem Cells (hDP-MSC) that increased in the presence of the bioactive HA signals. Moreover, in vivo studies via micro-CT and histological analyses of a bone model (e.g., a rat calvaria defect) confirmed that the maximum osteogenic response after 90 days was achieved with MAS30, which supported good regeneration of the calvaria defect without any evidence of inflammatory reaction. Hence, all of the results suggested that MAS is a promising scaffold for supporting the regeneration of hard tissues in different body compartments.
RESUMO
In recent years, ever-increasing scientific knowledge and modern high-tech advancements in micro- and nano-scales fabrication technologies have impacted significantly on various scientific fields. A micro-level approach so-called "microfluidic technology" has rapidly evolved as a powerful tool for numerous applications with special reference to bioengineering and biomedical engineering research. Therefore, a transformative effect has been felt, for instance, in biological sample handling, analyte sensing cell-based assay, tissue engineering, molecular diagnostics, and drug screening, etc. Besides such huge multi-functional potentialities, microfluidic technology also offers the opportunity to mimic different organs to address the complexity of animal-based testing models effectively. The combination of fluid physics along with three-dimensional (3-D) cell compartmentalization has sustained popularity as organ-on-a-chip. In this context, simple humanoid model systems which are important for a wide range of research fields rely on the development of a microfluidic system. The basic idea is to provide an artificial testing subject that resembles the human body in every aspect. For instance, drug testing in the pharma industry is crucial to assure proper function. Development of microfluidic-based technology bridges the gap between in vitro and in vivo models offering new approaches to research in medicine, biology, and pharmacology, among others. This is also because microfluidic-based 3-D niche has enormous potential to accommodate cells/tissues to create a physiologically relevant environment, thus, bridge/fill in the gap between extensively studied animal models and human-based clinical trials. This review highlights principles, fabrication techniques, and recent progress of organs-on-chip research. Herein, we also point out some opportunities for microfluidic technology in the future research which is still infancy to accurately design, address and mimic the in vivo niche.
RESUMO
Paracoccidioidomycosis (PCM) is a systemic mycosis endemic in Latin America, caused by Paracoccidioides spp. A limited number of antifungal agents are available and the search for new compounds has increased. Additionally, nanostructured lipid system (NLS) has emmerged as an interesting strategy to carrier compounds for the treatment of mycosis. In this work, the antifungal efficacy and toxicity of dodecyl gallate (DOD) associated with a NLS was evaluated through in vitro and in vivo tests. DOD showed good in vitro antifungal activity and low toxicity in lung fibroblasts and zebrafish embryos, but no antifungal efficacy in infected mice, which may have been a result of low bioavailability. On the other hand, the association of DOD + NLS was beneficial and resulted in lower toxicity in lung fibroblasts and zebrafish embryos. In addition, NLSâ¯+â¯DOD promoted a significant reduction in the fungal burden of mice lungs and could be a potential therapeutic option against PCM.
Assuntos
Antifúngicos/farmacologia , Ácido Gálico/análogos & derivados , Nanopartículas/química , Paracoccidioides/efeitos dos fármacos , Paracoccidioidomicose/tratamento farmacológico , Animais , Antifúngicos/química , Antifúngicos/uso terapêutico , Disponibilidade Biológica , Linhagem Celular , Modelos Animais de Doenças , Feminino , Fibroblastos , Ácido Gálico/química , Ácido Gálico/farmacologia , Ácido Gálico/uso terapêutico , Humanos , Concentração Inibidora 50 , Lipídeos/química , Pulmão/citologia , Pulmão/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Paracoccidioides/isolamento & purificação , Paracoccidioidomicose/microbiologia , Resultado do Tratamento , Peixe-ZebraRESUMO
INTRODUCTION AND OBJECTIVES: Achalasia is the most widely studied esophageal motility disorder. No treatment has achieved completely satisfactory results. The laparoscopic Heller esophagomyotomy is currently the most accepted technique. With the advent of minimally invasive surgery, the appearance of peroral endoscopic myotomy (POEM) has promising results. The primary aim of our study was to perform peroral endoscopic esophagomyotomy in animal experimentation models to perfect the technique and later apply it to humans. The secondary aims were to evaluate the intraoperative and postoperative complications and to describe the anatomopathologic findings. MATERIALS AND METHODS: An experimental study was conducted on 8 live porcine models that were followed for 30 days to identify postoperative complications. Necropsy was then performed to evaluate the histopathologic findings. The international requirements and regulations for animal experimentation were met. RESULTS: The technique was carried out in all the models. There was one intraoperative death. Pneumothorax was observed in 50% of the units in experimentation and subcutaneous cervical emphysema in 75%, with no significant clinical repercussions. Histologic muscle layer (myotomy) involvement was above the gastroesophageal junction in 87% of the cases and below it in 25%. CONCLUSION: Peroral endoscopic esophagomyotomy is a feasible, albeit complex, procedure that requires advanced training, and thus should be performed in highly specialized centers. Specific skills in advanced therapeutic endoscopic procedures of this type must continue to be developed through continuing education (ideally in in vivo models), to then be performed on humans.
Assuntos
Acalasia Esofágica/cirurgia , Miotomia de Heller/métodos , Cirurgia Endoscópica por Orifício Natural/métodos , Animais , Feminino , Masculino , Complicações Pós-Operatórias/diagnóstico , Suínos , Resultado do TratamentoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Ipomoea asarifolia (Desr.) Roem. and Schult.(Convolvulaceae), popularly known as salsa or salsa-brava, is a plant of which the decoction of leaves is used in folk medicine to treat various inflammatory disorders such of dermatitis, scabies, symptoms of syphilis, skin ulcers and external wounds. However, little is known about possible compounds and mechanisms of action of the plant to support the activities reported by popular use. AIM OF THE STUDY: The study aimed to identify bioactive molecules present in the crude extract of I. asarifolia leaves and investigate the anti-inflammatory potential of this extract in different experimental in vivo models to improve the understanding on that activity. MATERIAL AND METHODS: Aqueous extract of I. asarifolia leaves was prepared by decoction (1:10 m/v) and its chromatographic profile was obtained by high performance liquid chromatography coupled with diode array detector (HPLC-DAD) and liquid chromatography diode array detector coupled with mass spectrometry (LC-DAD-MS). The potential anti-inflammatory activity of the extract was assessed using the following in vivo models: xylene-induced ear edema (20, 30 and 40mg/kg), evaluating the degree of edema formation; carrageenan-induced peritonitis (10, 20 and 30mg/kg), evaluating leukocyte migration and cytokine levels (IL-1ß, IL-6, IL-12 and TNF-α) at 4h; zymosan-induced air pouch inflammation (20, 30 and 40mg/kg), evaluating the kinetics of leukocyte migration by total and differential counts at 6, 24 and 48h. The same tests were conducted using pure compounds identified in the aqueous extract from I. asarifolia leaves in different doses for each experimental model. RESULTS: The compounds identified in the aqueous extract of I. asarifolia leaves by HPLC-DAD and LC-DAD-MS were rutin, chlorogenic acid and caffeic acid. The extract significantly reduced ear edema induced by xylene (81%, 85% and 86% for doses of 20, 30 and 40mg/kg, respectively, p<0.001), as well as cell migration in experimental models of peritonitis (70%, 78% and 83% for doses of 10, 20 and 30mg/kg, respectively, p<0.001) and air pouch inflammation (58%, 67% and 53% for doses of 20, 30 and 40mg/kg, respectively, p<0.001). In addition, the extract demonstrated the ability to significantly inhibit the production of cytokines IL-1ß, IL-6, IL-12 and TNF-α (p<0.001). The secondary metabolites tested (rutin, chlorogenic acid and caffeic acid) also showed the ability to significantly (p<0.001) decrease the parameters analyzed above. CONCLUSION: This is the first study to identify and confirm these phenolic compounds in I. asarifolia leaves extract and to suggest that these compounds contribute to the anti-inflammatory activity in vivo, as reported by ethnomedicinal use of this plant. Through the different experimental models performed, we can conclude that the results obtained with the aqueous extract from I. asarifolia leaves support its popular use for the treatment of inflammatory disorders.
Assuntos
Anti-Inflamatórios/farmacologia , Edema/prevenção & controle , Inflamação/prevenção & controle , Ipomoea/química , Peritonite/prevenção & controle , Fenóis/farmacologia , Extratos Vegetais/farmacologia , Folhas de Planta/química , Animais , Anti-Inflamatórios/isolamento & purificação , Carragenina , Quimiotaxia de Leucócito/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Citocinas/metabolismo , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Edema/induzido quimicamente , Edema/metabolismo , Feminino , Inflamação/induzido quimicamente , Inflamação/metabolismo , Mediadores da Inflamação/metabolismo , Contagem de Leucócitos , Leucócitos/efeitos dos fármacos , Leucócitos/metabolismo , Masculino , Espectrometria de Massas , Camundongos Endogâmicos BALB C , Peritonite/induzido quimicamente , Peritonite/metabolismo , Fenóis/isolamento & purificação , Fitoterapia , Extratos Vegetais/isolamento & purificação , Plantas Medicinais , Fatores de Tempo , Xilenos , ZimosanRESUMO
Chemokines are essential mediators of leukocyte movement in vivo. In vitro assays of leukocyte migration cannot mimic the complex interactions with other cell types and matrix needed for cells to extravasate and migrate into tissues. Therefore, in vivo strategies to study the effects and potential relevance of chemokines for the migration of particular leukocyte subsets are necessary. Here, we describe methods to study the effects and endogenous role of chemokine in mice. Advantages and pitfalls of particular models are discussed and we focus on description in model's joint and pleural cavity inflammation and the effects and relevance of CXCR2 and CCR2 ligands on cell migration.
Assuntos
Artrite Experimental/metabolismo , Quimiocinas/metabolismo , Quimiotaxia de Leucócito , Animais , Movimento Celular , Quimiocina CCL2/metabolismo , Quimiocina CXCL1/metabolismo , Articulações/patologia , Camundongos , Microscopia Confocal/métodos , Neutrófilos/metabolismo , Neutrófilos/patologia , Cavidade Pleural/metabolismo , Cavidade Pleural/patologia , Receptores CCR2/metabolismo , Receptores de Interleucina-8B/administração & dosagem , Receptores de Interleucina-8B/metabolismoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Cancer cases numbers are increasing worldwide positioning this disease as the second cause of mortality for both sexes. Medicinal plants have been used in the fight against cancer as the basis for drug discovery and nowadays more than 70% of anticancer drugs have a natural origin. Mexico is regarded for its cultural and biological diversity, which is reflected in the vast traditional knowledge of herbal remedies. In this review we examined herbal remedies employed in colorectal cancer treatment (CRC). AIM OF THE STUDY: The goal of this work was to gather scientific reports of plants used in Mexican traditional medicine for CRC treatment. MATERIALS AND METHODS: We performed a search on scientific literature databases using as keywords: "colon cancer", "gastric cancer", "cytotoxicity", studies "in vitro and in vivo", in combination with "Mexican medicinal plants" or "Mexican herbal remedies". The selection criteria of cytotoxic activity for extracts or pure compounds was based on the National Cancer Institute of USA recommendations of effective dose 50 (ED50) of ≤20µg/mL and ≤4µg/mL, respectively. RESULTS: In this review we report 25 botanic families and 39 species of plants used for the treatment of colon cancer in Mexico with evidence in studies in vitro and in vivo. CONCLUSIONS: Medicinal plants are still a great source of novel chemical structures with antineoplastic potential as it is proven in this work. The selection criteria and activity was narrowed for methodological purposes, nevertheless, drug discovery of natural origin continues to be a highly attractive R&D strategy.