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1.
J Nutr Biochem ; 101: 108925, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-34843933

RESUMO

Maternal high-fat diet (HFD) often results in intrauterine and feto-placental inflammation, and increases the risks of fetal programming of metabolic diseases. Intake of prebiotic is reported beneficial. However, its effects on HFD during pregnancy and lactation is not known. We evaluated the maternal intake of fructooligosaccharide (FOS) and its impact on placental inflammation, offspring's adiposity, glucose, and lipid metabolism in their later life. Female Golden Syrian hamsters were fed with a control diet (CD, 26.4 % energy from fat) or HFD (60.7% energy from fat) in the presence or absence of FOS from preconception until lactation. All pups were switched over to CD after lactation and continued until the end. Placental inflammation was upregulated in HFD-fed dam, as measured by a high concentration of hsCRP in the serum and amniotic fluid. Neutrophil infiltration was significantly increased in the decidua through the chorionic layer of the placenta. The expression of pro-inflammatory cytokines such as COX2, NFκß, IL-8, TGFß mRNA was increased in the chorioamniotic membrane (P <.05). The HFD/CD hamsters had more adiposity, higher triglyceride, and low HDL at 12 months of age compared to CD/CD (P <.05). However, HFD+FOS/CD-fed hamsters prevented adverse effects such as placental inflammation, neutrophil infiltration, glucose, and lipid profiles in the offspring (P <.05). Anti-inflammatory and lipid-lowering effects of FOS may reduce placental inflammation by lowering neutrophil infiltration and decreasing the production of pro-inflammatory cytokines. Intake of FOS during pregnancy may be beneficial in maintaining lipid metabolism and preventing excess adiposity for mother and their offspring.


Assuntos
Dieta Hiperlipídica/efeitos adversos , Inflamação/prevenção & controle , Lipídeos/sangue , Fenômenos Fisiológicos da Nutrição Materna , Oligossacarídeos , Prebióticos , Adiposidade , Animais , Glicemia/análise , Peso Corporal , Membrana Corioalantoide/imunologia , Citocinas/metabolismo , Feminino , Metabolismo dos Lipídeos , Lipoproteínas HDL/sangue , Mesocricetus , Infiltração de Neutrófilos , Placenta/imunologia , Gravidez , Triglicerídeos/sangue
2.
Front Immunol ; 12: 791081, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34868080

RESUMO

Dysregulation of the immune system is associated with many pathologies, including cardiovascular diseases, diabetes, and cancer. To date, the most commonly used models in biomedical research are rodents, and despite the various advantages they offer, their use also raises numerous drawbacks. Recently, another in vivo model, the chicken embryo and its chorioallantoic membrane, has re-emerged for various applications. This model has many benefits compared to other classical models, as it is cost-effective, time-efficient, and easier to use. In this review, we explain how the chicken embryo can be used as a model for immune-based studies, as it gradually develops an embryonic immune system, yet which is functionally similar to humans'. We mainly aim to describe the avian immune system, highlighting the differences and similarities with the human immune system, including the repertoire of lymphoid tissues, immune cells, and other key features. We also describe the general in ovo immune ontogeny. In conclusion, we expect that this review will help future studies better tailor their use of the chicken embryo model for testing specific experimental hypotheses or performing preclinical testing.


Assuntos
Embrião de Galinha/imunologia , Membrana Corioalantoide/imunologia , Sistema Imunitário/imunologia , Animais , Embrião de Galinha/metabolismo , Membrana Corioalantoide/metabolismo , Citocinas/genética , Citocinas/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Sistema Imunitário/crescimento & desenvolvimento , Sistema Imunitário/metabolismo , Mediadores da Inflamação/metabolismo , Modelos Animais , Receptores Imunológicos/genética , Receptores Imunológicos/metabolismo , Transdução de Sinais , Especificidade da Espécie
3.
Infect Immun ; 89(5)2021 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-33558326

RESUMO

Preterm labor precedes premature birth, the leading cause of neonatal morbidity and mortality worldwide. Preterm labor can occur in the context of either microbe-associated intra-amniotic inflammation (i.e., intra-amniotic infection) or intra-amniotic inflammation in the absence of detectable microorganisms (i.e., sterile intra-amniotic inflammation). Both intra-amniotic infection and sterile intra-amniotic inflammation trigger local immune responses that have deleterious effects on fetal life. Yet, the extent of such immune responses in the fetal tissues surrounding the amniotic cavity (i.e., the chorioamniotic membranes) is poorly understood. By using RNA sequencing (RNA seq) as a discovery approach, we found that there were significant transcriptomic differences involving host response to pathogens in the chorioamniotic membranes of women with intra-amniotic infection compared to those from women without inflammation. In addition, the sterile or microbial nature of intra-amniotic inflammation was associated with distinct transcriptomic profiles in the chorioamniotic membranes. Moreover, the immune response in the chorioamniotic membranes of women with sterile intra-amniotic inflammation was milder in nature than that induced by microbes and involved the upregulation of alarmins and inflammasome-related molecules. Lastly, the presence of maternal and fetal inflammatory responses in the placenta was associated with the upregulation of immune processes in the chorioamniotic membranes. Collectively, these findings provide insight into the immune responses against microbes or alarmins that take place in the fetal tissues surrounding the amniotic cavity, shedding light on the immunobiology of preterm labor and birth.


Assuntos
Membrana Corioalantoide/imunologia , Membrana Corioalantoide/microbiologia , Inflamação/etiologia , Trabalho de Parto Prematuro/etiologia , Feminino , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Inflamassomos/metabolismo , Inflamação/metabolismo , Trabalho de Parto Prematuro/metabolismo , Gravidez , Análise de Sequência de RNA , Transcriptoma
4.
Int J Mol Sci ; 21(20)2020 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-33066403

RESUMO

The effective management of tissue integration and immunological responses to transplants decisively co-determines the success of soft and hard tissue reconstruction. The aim of this in vivo study was to evaluate the eligibility of extracorporeal shock wave therapy (ESWT) with respect to its ability to modulate angiogenesis and immune response to a collagen matrix (CM) for tissue engineering in the chorioallantoic membrane (CAM) assay, which is performed with fertilized chicken eggs. CM were placed on the CAM on embryonic development day (EDD) 7; at EDD-10, ESWT was conducted at 0.12 mJ/mm2 with 500 impulses each. One and four days later, angiogenesis represented by vascularized area, vessel density, and vessel junctions as well as HIF-1α and VEGF gene expression were evaluated. Furthermore, immune response (iNOS2, MMP-9, and MMP-13 via qPCR) was assessed and compared between ESWT- and non-ESWT-groups. At EDD-14, the vascularized area (+115% vs. +26%) and the increase in vessel junctions (+751% vs. +363%) were significantly higher in the ESWT-group. ESWT significantly increased MMP-9 gene expression at EDD-11 and significantly decreased MMP-13 gene expression at EDD-14 as compared to the controls. Using the CAM assay, an enhanced angiogenesis and neovascularization in CM after ESWT were observed. Furthermore, ESWT could reduce the inflammatory activity after a latency of four days.


Assuntos
Colágeno/imunologia , Tratamento por Ondas de Choque Extracorpóreas/métodos , Neovascularização Fisiológica , Engenharia Tecidual/métodos , Animais , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Embrião de Galinha , Membrana Corioalantoide/irrigação sanguínea , Membrana Corioalantoide/imunologia , Colágeno/química , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Metaloproteinases da Matriz/genética , Metaloproteinases da Matriz/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Alicerces Teciduais/efeitos adversos , Alicerces Teciduais/química , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo
5.
Front Immunol ; 11: 581288, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33117388

RESUMO

Pathological angiogenesis of the retina is a key component of irreversible causes of blindness, as observed in proliferative diabetic retinopathy (PDR). The pathogenesis of PDR is complex and involves vascular, inflammatory, and neuronal mechanisms. Several structural and molecular alterations associated to PDR are related to the presence of inflammation that appears to play a non-redundant role in the neovascular response that characterizes the retina of PDR patients. Vascular endothelial growth factor (VEGF) blockers have evolved over time for the treatment of retinal neovascularization. However, several limitations to anti-VEGF interventions exist. Indeed, the production of other angiogenic factors and pro-inflammatory mediators may nullify and/or cause resistance to anti-VEGF therapies. Thus, appropriate experimental models are crucial for dissecting the mechanisms leading to retinal neovascularization and for the discovery of more efficacious anti-angiogenic/anti-inflammatory therapies for PDR patients. This review focuses on the tight cross talk between angiogenesis and inflammation during PDR and describe how the chick embryo chorioallantoic membrane (CAM) assay may represent a cost-effective and rapid in vivo tool for the study of the relationship between neovascular and inflammatory responses elicited by the vitreous humor of PDR patients and for the screening of novel therapeutic agents.


Assuntos
Retinopatia Diabética/etiologia , Neovascularização Retiniana/etiologia , Inibidores da Angiogênese/uso terapêutico , Animais , Embrião de Galinha , Membrana Corioalantoide/irrigação sanguínea , Membrana Corioalantoide/imunologia , Membrana Corioalantoide/patologia , Retinopatia Diabética/imunologia , Retinopatia Diabética/patologia , Humanos , Inflamação/etiologia , Inflamação/imunologia , Inflamação/patologia , Modelos Animais , Modelos Biológicos , Neovascularização Retiniana/imunologia , Neovascularização Retiniana/patologia , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Fator A de Crescimento do Endotélio Vascular/fisiologia , Corpo Vítreo/imunologia , Corpo Vítreo/patologia
6.
Int J Mol Sci ; 20(15)2019 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-31357391

RESUMO

During pregnancy, the placenta, the mother and the fetus exploit several mechanisms in order to avoid fetal rejection and to maintain an immunotolerant environment throughout nine months. During this time, immune cells from the fetal and maternal compartments interact to provide an adequate defense in case of an infection and to promote a tolerogenic milieu for the fetus to develop peacefully. Trophoblasts and decidual cells, together with resident natural killer cells, dendritic cells, Hofbauer cells and other macrophages, among other cell types, contribute to the modulation of the uterine environment to sustain a successful pregnancy. In this review, the authors outlined some of the various roles that the innate immune system plays at the maternal-fetal interface. First, the cell populations that are recruited into gestational tissues and their immune mechanisms were examined. In the second part, the Toll-like receptor (TLR)-dependent immune responses at the maternal-fetal interface was summarized, in terms of their specific cytokine/chemokine/antimicrobial peptide expression profiles throughout pregnancy.


Assuntos
Imunidade Inata , Imunidade , Troca Materno-Fetal , Receptores Toll-Like/metabolismo , Animais , Biomarcadores , Membrana Corioalantoide/imunologia , Citocinas/metabolismo , Feminino , Humanos , Imunomodulação , Placenta/imunologia , Placenta/metabolismo , Gravidez
7.
J Vet Sci ; 20(1): 51-57, 2019 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-30481981

RESUMO

Monoclonal antibodies (MAbs) are widely applied in disease diagnoses. Herein, we report a MAb, WF-4, against Influenza A virus nucleoprotein (NP), its broad response with Influenza A virus, and its application in an immunohistochemistry (IHC) assay. WF-4 was screened by immunofluorescence assay (IFA). The results showed that its reactivity with baculovirus-expressed full-length recombinant NP (rNP) in Western blot (WB), indicating its IHC applicability. Fifteen Influenza A virus (reference subtypes H1 to H15) infected chicken embryonated chorioallantoic membranes (CAM), fixed by formalin, were all detectable in the WF-4-based IHC assay. Also, the reactivity of the IHC test with NP from experimentally inoculated H6N1 and from all recent outbreaks of H5 subtype avian Influenza A virus (AIV) field cases in Taiwan showed positive results. Our data indicate that CAM, a by-product of Influenza A virus preparation, is helpful for Influenza A virus-specific MAb characterization, and that the WF-4 MAb recognizes conserved and linear epitopes of Influenza A virus NP. Therefore, WF-4 is capable of detecting NP antigens via IHC and may be suitable for developing various tests for diagnosis of Influenza A virus and, especially, AIV infection.


Assuntos
Galinhas , Imuno-Histoquímica/veterinária , Vírus da Influenza A Subtipo H5N2/imunologia , Vírus da Influenza A/imunologia , Proteínas de Ligação a RNA/imunologia , Proteínas do Core Viral/imunologia , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Antivirais/imunologia , Membrana Corioalantoide/imunologia , Imunofluorescência/veterinária , Proteínas do Nucleocapsídeo , Taiwan
8.
In Vivo ; 32(4): 791-798, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29936460

RESUMO

AIM: To characterize baby hamster kidney fibroblast (BHK 21/C13) cells and test the effects of antibodies against podoplanin and disodium cromolyn on BHK 21/C13 cell line-derived tumors grown on chick embryo chorioallantoic membrane (CAM). MATERIAL AND METHODS: BHK 21/C13 cell-derived fibrosarcomas developed in hamsters were implanted on CAM and treated with anti-podoplanin antibodies and disodium cromolyn. BHK 21/C13 cell immunophenotype was assessed. RESULTS: Fibrosarcoma cells were positive for vimentin, CD117, smooth muscle actin, vascular endothelial growth factor epidermal growth factor receptor, homebox prospero gene 1 and negative for platelet-derived growth factor B, neuron-specific enolase, S100, CD34, Ewing sarcoma and podoplanin. CAM-grown fibrosarcomas were highly sensitive to disodium cromolyn and anti-podoplanin antibodies. CONCLUSION: Immunophenotyping BHK 21/C13 cells and their response to drugs represent the first step in revealing cell line utility and a reliable tool for experimental cancer research.


Assuntos
Membrana Corioalantoide/efeitos dos fármacos , Fibrossarcoma/tratamento farmacológico , Glicoproteínas de Membrana/antagonistas & inibidores , Proteínas de Neoplasias/genética , Animais , Anticorpos Anti-Idiotípicos/administração & dosagem , Linhagem Celular Tumoral , Embrião de Galinha , Membrana Corioalantoide/imunologia , Cricetinae , Cromolina Sódica/administração & dosagem , Fibroblastos/efeitos dos fármacos , Fibroblastos/imunologia , Fibrossarcoma/genética , Fibrossarcoma/imunologia , Fibrossarcoma/patologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glicoproteínas de Membrana/imunologia , Proteínas Proto-Oncogênicas c-kit/genética , Vimentina/genética
9.
Arch Immunol Ther Exp (Warsz) ; 64(6): 515-522, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27289241

RESUMO

Scattered data suggested that disodium cromolyn, well known as a mast cell stabilizer shows some effects on tumor cells and tumor-associated newly formed vascular networks. Most of these studies used tumor cell lines assessed by in vitro studies. Nor disodium cromolyn effects on melanoma cell lines were studied yet, neither its influence on recruited tumor blood vessels or angiogenic growth factors expression. We designed here a study regarding disodium cromolyn effects on A375 melanoma tumor cells implanted on chick embryo chorioallantoic membrane (CAM) and on blood vessels recruited by the experimental melanoma in the absence of mast cells, knowing that within CAM, the existence of mast cells are not certified yet. We also assessed the role of disodium cromolyn on the expression of several angiogenic growth factors. Disodium cromoglycate differentially acts on tumor cells and blood vessels. Extensive necrotic areas of experimental melanoma together with an increased number of peritumor blood vessels were observed in treated specimens as compared with untreated tumors. Disodium cromolyn inhibited VEGF and PDGF-BB expression, and had no effects on EG VEGF expression between treated and non treated specimens in a mast cells free microenvironment. Our results sustain the direct antitumor effects of sodium cromolyn and suggest the involvement of several growth factors in the recruitment of tumor vessels by A375 melanoma tumor cells. The expression of growth factors is differentially influenced by sodium cromolyn treatment.


Assuntos
Indutores da Angiogênese/química , Antineoplásicos/imunologia , Membrana Corioalantoide/imunologia , Cromolina Sódica/química , Mastócitos/efeitos dos fármacos , Mastócitos/imunologia , Neovascularização Patológica , Animais , Antiasmáticos/química , Becaplermina , Vasos Sanguíneos/imunologia , Linhagem Celular Tumoral , Embrião de Galinha , Feminino , Humanos , Mastócitos/citologia , Melanoma/imunologia , Pessoa de Meia-Idade , Necrose , Proteínas Proto-Oncogênicas c-sis/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo
12.
J Biomater Sci Polym Ed ; 26(13): 828-40, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26063500

RESUMO

Poly(lactic-co-glycolic acid) (PLGA) microspheres have been widely used as drug carriers for minimally invasive, local, and sustained drug delivery. However, their use is often plagued by limited controllability of encapsulation efficiency, initial burst, and release rate of drug molecules, which cause unsatisfactory outcomes and several side effects including inflammation. This study presents a new strategy of tuning the encapsulation efficiency and the release rate of protein drugs from a PLGA microsphere by filling the hollow core of the microsphere with poly(ethylene glycol) (PEG) hydrogels of varying cross-linking density. The PEG gel cores were prepared by inducing in situ cross-linking reactions of PEG monoacrylate solution within the PLGA microspheres. The resulting PEG-PLGA core-shell microspheres exhibited (1) increased encapsulation efficiency, (2) decreased initial burst, and (3) a more sustained release of protein drugs, as the cross-linking density of the PEG gel core was increased. In addition, implantation of PEG-PLGA core-shell microspheres encapsulated with vascular endothelial growth factor (VEGF) onto a chicken chorioallantoic membrane resulted in a significant increase in the number of new blood vessels at an implantation site, while minimizing inflammation. Overall, this strategy of introducing PEG gel into PLGA microspheres will be highly useful in tuning release rates and ultimately in improving the therapeutic efficacy of a wide array of protein drugs.


Assuntos
Sistemas de Liberação de Medicamentos/métodos , Microesferas , Poliésteres , Polietilenoglicóis , Indutores da Angiogênese/administração & dosagem , Animais , Galinhas , Membrana Corioalantoide/irrigação sanguínea , Membrana Corioalantoide/imunologia , Membrana Corioalantoide/cirurgia , Implantes de Medicamento/química , Liberação Controlada de Fármacos , Hidrogéis/química , Poliésteres/química , Polietilenoglicóis/química , Proteínas/administração & dosagem , Soroalbumina Bovina/metabolismo , Fator A de Crescimento do Endotélio Vascular/administração & dosagem
13.
Viral Immunol ; 28(4): 192-200, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25874816

RESUMO

Early responses against viruses, such as avian influenza virus (AIV), may be induced by Toll-like receptor (TLR) pathways. In the present study, an in ovo model was employed to study the antiviral activities of TLR ligands. It was hypothesized that administration of TLR ligands in ovo at the appropriate dose and time can reduce AIV titer in embryonated chicken eggs. Moreover, the study aimed to determine the mechanisms involved in the TLR-mediated antiviral responses in the chorioallantoic membrane (CAM). Embryonated eggs (10-14 day old) were treated with TLR2, 4, 7, and 21 ligands using different doses and times pre- and post-AIV infection. The results revealed that treatment of embryonated chicken eggs with TLR ligands reduced AIV replication. Further analysis showed that TLR ligands induced interferon (IFN)-γ and IFN stimulatory genes in the CAM, which may have played a role in the reduction of the AIV titer. The timing and dose of TLR ligands administration had significant impacts on the outcome of the treated eggs. In conclusion, the present study demonstrated that the in ovo route may be employed to determine the antiviral characteristics of TLR ligands against AIV.


Assuntos
Membrana Corioalantoide/imunologia , Fatores Imunológicos/metabolismo , Orthomyxoviridae/crescimento & desenvolvimento , Orthomyxoviridae/imunologia , Receptores Toll-Like/agonistas , Receptores Toll-Like/metabolismo , Animais , Embrião de Galinha , Interferons/metabolismo , Carga Viral , Replicação Viral
14.
PLoS One ; 6(5): e19741, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21603634

RESUMO

Alternative models of microbial infections are increasingly used to screen virulence determinants of pathogens. In this study, we investigated the pathogenesis of Candida albicans and C. glabrata infections in chicken embryos infected via the chorio-allantoic membrane (CAM) and analyzed the virulence of deletion mutants. The developing immune system of the host significantly influenced susceptibility: With increasing age, embryos became more resistant and mounted a more balanced immune response, characterized by lower induction of proinflammatory cytokines and increased transcription of regulatory cytokines, suggesting that immunopathology contributes to pathogenesis. While many aspects of the chicken embryo response resembled murine infections, we also observed significant differences: In contrast to systemic infections in mice, IL-10 had a beneficial effect in chicken embryos. IL-22 and IL-17A were only upregulated after the peak mortality in the chicken embryo model occurred; thus, the role of the Th17 response in this model remains unclear. Abscess formation occurs frequently in murine models, whereas the avian response was dominated by granuloma formation. Pathogenicity of the majority of 15 tested C. albicans deletion strains was comparable to the virulence in mouse models and reduced virulence was associated with significantly lower transcription of proinflammatory cytokines. However, fungal burden did not correlate with virulence and for few mutants like bcr1Δ and tec1Δ different outcomes in survival compared to murine infections were observed. C. albicans strains locked in the yeast stage disseminated significantly more often from the CAM into the embryo, supporting the hypothesis that the yeast morphology is responsible for dissemination in systemic infections. These data suggest that the pathogenesis of C. albicans infections in the chicken embryo model resembles systemic murine infections but also differs in some aspects. Despite its limitations, it presents a useful alternative tool to pre-screen C. albicans strains to select strains for subsequent testing in murine models.


Assuntos
Candidíase/imunologia , Membrana Corioalantoide/microbiologia , Modelos Animais de Doenças , Animais , Candida albicans/patogenicidade , Candida glabrata/patogenicidade , Embrião de Galinha , Membrana Corioalantoide/imunologia , Citocinas/biossíntese , Citocinas/genética , Regulação da Expressão Gênica , Inflamação , Camundongos
15.
Exp Dermatol ; 18(4): 409-13, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19054059

RESUMO

The current standard method for predicting contact allergenicity is the murine local lymph node assay (LLNA). Public objection to the use of animals in testing of cosmetics makes the development of a system that does not use sentient animals highly desirable. The chorioallantoic membrane (CAM) of the chick egg has been extensively used for the growth of normal and transformed mammalian tissues. The CAM is not innervated, and embryos are sacrificed before the development of pain perception. The aim of this study was to determine whether the sensitization phase of contact dermatitis to known cosmetic allergens can be quantified using CAM-engrafted human skin and how these results compare with published EC3 data obtained with the LLNA. We studied six common molecules used in allergen testing and quantified migration of epidermal Langerhans cells (LC) as a measure of their allergic potency. All agents with known allergic potential induced statistically significant migration of LC. The data obtained correlated well with published data for these allergens generated using the LLNA test. The human-skin CAM model therefore has great potential as an inexpensive, non-radioactive, in vivo alternative to the LLNA, which does not require the use of sentient animals. In addition, this system has the advantage of testing the allergic response of human, rather than animal skin.


Assuntos
Alérgenos/imunologia , Membrana Corioalantoide/imunologia , Cosméticos/efeitos adversos , Dermatite Alérgica de Contato/imunologia , Modelos Biológicos , Pele/imunologia , Animais , Movimento Celular/efeitos dos fármacos , Embrião de Galinha , Cosméticos/farmacologia , Dermatite Alérgica de Contato/etiologia , Humanos , Células de Langerhans/patologia , Modelos Animais , Valor Preditivo dos Testes , Testes Cutâneos , Transplantes
16.
Biomaterials ; 30(4): 622-31, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18996588

RESUMO

In this study, we examined the in vivo pharmacological effects of the sequential delivery of dexamethasone (DX) followed by vascular endothelial growth factor (VEGF) on the immune response and localized vascular network formation around a hydrogel-coated, micro-capillary implant for single-walled carbon nanotube based fluorescence sensors. We demonstrate, for the first time, imaging of an SWNT fluorescence device implanted subcutaneously in a rat. For tissue response studies, the chick embryo chorioallantoic membrane (CAM) was used as a tissue-model for an 8-day implantation period. The average vascular density of the tissue surrounding a hydrogel-coated microdialysis capillary sensor with simultaneous, sequential, or no delivery of DX and VEGF was 1.24+/-0.35x10(-3)vessels/microm(2), 1.15+/-0.30x10(-3)vessels/microm(2) and 0.71+/-0.20x10(-3)vessels/microm(2), respectively. Calculation of the therapeutic index (vasculature/inflammation ratio), which reflects promotion of angiogenesis versus the host immune response, demonstrates that sequential DX/VEGF delivery was 60.3% and 139.3% higher than that of VEGF and DX release alone, respectively, and was also 32.1% higher when compared to simultaneous administration, proving to be a more effective strategy in utilizing the pharmacological impact of DX and VEGF around the biosensor-model implant.


Assuntos
Técnicas Biossensoriais , Prótese Vascular , Capilares/metabolismo , Membrana Corioalantoide/irrigação sanguínea , Dexametasona/administração & dosagem , Sistemas de Liberação de Medicamentos/métodos , Nanotubos de Carbono , Fator A de Crescimento do Endotélio Vascular/administração & dosagem , Actinas/metabolismo , Animais , Capilares/citologia , Capilares/efeitos dos fármacos , Embrião de Galinha , Membrana Corioalantoide/citologia , Membrana Corioalantoide/efeitos dos fármacos , Membrana Corioalantoide/imunologia , DNA/metabolismo , Dexametasona/farmacologia , Fluorescência , Imunidade/efeitos dos fármacos , Inflamação , Neovascularização Fisiológica/efeitos dos fármacos , Polietilenoglicóis , Ratos , Fator A de Crescimento do Endotélio Vascular/farmacologia
17.
Histochem Cell Biol ; 130(6): 1119-30, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19005674

RESUMO

Since their introduction almost a century ago, chick embryo model systems involving the technique of chorioallantoic grafting have proved invaluable in the in vivo studies of tumor development and angiogenesis and tumor cell dissemination. The ability of the chick embryo's chorioallantoic membrane (CAM) to efficiently support the growth of inoculated xenogenic tumor cells greatly facilitates analysis of human tumor cell metastasis. During spontaneous metastasis, the highly vascularized CAM sustains rapid tumor formation within several days following cell grafting. The dense capillary network of the CAM also serves as a repository of aggressive tumor cells that escaped from the primary tumor and intravasated into the host vasculature. This spontaneous metastasis setting provides a unique experimental model to study in vivo the intravasation step of the metastatic cascade. During experimental metastasis when tumor cells are inoculated intravenously, the CAM capillary system serves as a place for initial arrest and then, for tumor cell extravasation and colonization. The tissue composition and accessibility of the CAM for experimental interventions makes chick embryo CAM systems attractive models to follow the fate and visualize microscopically the behavior of grafted tumor cells in both spontaneous and experimental metastasis settings.


Assuntos
Membrana Corioalantoide/irrigação sanguínea , Neoplasias/patologia , Animais , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Embrião de Galinha , Membrana Corioalantoide/imunologia , Humanos , Microscopia/métodos , Modelos Animais , Monócitos/imunologia , Invasividade Neoplásica , Metástase Neoplásica , Neoplasias/irrigação sanguínea , Neoplasias/imunologia , Células Neoplásicas Circulantes/patologia , Neovascularização Fisiológica
18.
Cancer Cell ; 13(3): 221-34, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18328426

RESUMO

In vivo tumor cell migration through integrin-dependent pathways is key to the metastatic behavior of malignant cells. Using quantitative in vivo assays and intravital imaging, we assessed the impact of cell migration, regulated by the integrin-associated tetraspanin CD151, on spontaneous human tumor cell metastasis. We demonstrate that promoting immobility through a CD151-specific metastasis blocking mAb prevents tumor cell dissemination by inhibiting intravasation without affecting primary tumor growth, tumor cell arrest, extravasation, or growth at the secondary site. In vivo, this loss of migration is the result of enhanced tumor cell-matrix interactions, promoted by CD151, which prevent dissociation by individual cells and leads to a subsequent inhibition of invasion and intravasation at the site of the primary tumor.


Assuntos
Antígenos CD/metabolismo , Movimento Celular , Membrana Corioalantoide/imunologia , Neoplasias Experimentais/imunologia , Animais , Anticorpos Monoclonais , Antígenos CD/genética , Antígenos CD/imunologia , Sítios de Ligação de Anticorpos , Adesão Celular , Linhagem Celular Tumoral , Proliferação de Células , Embrião de Galinha , Membrana Corioalantoide/irrigação sanguínea , Membrana Corioalantoide/metabolismo , Membrana Corioalantoide/patologia , Matriz Extracelular/metabolismo , Humanos , Camundongos , Camundongos Knockout , Camundongos SCID , Microscopia de Fluorescência , Microscopia de Vídeo , Invasividade Neoplásica , Metástase Neoplásica , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/patologia , Reação em Cadeia da Polimerase/métodos , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Tetraspanina 24 , Fatores de Tempo , Transfecção
19.
Artigo em Inglês | MEDLINE | ID: mdl-17969440

RESUMO

The collaboration between Alick Isaacs and myself started in the summer of 1956. Our initial project was to show, by electron microscopy, that interference between inactivated influenza virus and live virus involved the transfer of material from the interfering virus to the host cell. This approach failed for technical reasons. However, in the course of this work it appeared that more interfering activity remained in the system than we were entitled to expect. One possible explanation was that a substance, not identical with the initial interfering virus, was being generated. Subsequent experiments, aimed at checking this hypothesis, led to the description of interferon.


Assuntos
Interferons/história , Animais , Membrana Corioalantoide/imunologia , Membrana Corioalantoide/virologia , História do Século XX , Vírus da Influenza A/imunologia , Interferons/imunologia
20.
Cutan Ocul Toxicol ; 26(1): 25-36, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17464746

RESUMO

A large database has demonstrated a robust relationship between the chorioallantonic membrane (CAM) and the Draize rabbit eye tests. The precision, sensitivity, and direct correlation of the CAM with human eye evaluations were critically appraised in this work. A total of 24 studies, including 12 CAM assays and 12 human clinical studies were conducted. Both the CAM assay and the human tests demonstrated good interassay precision and a robust correlation between the CAM scores and the inflammatory responses of human eye bulbar conjunctiva, palpebral conjunctiva and scleral vessels. This provided sound evidence that the CAM assay could be a predictive screen for eye irritation assessment for personal wash products and a key element of the tiered scientific approach in supporting labeling claims such as "tear free".


Assuntos
Membrana Corioalantoide/efeitos dos fármacos , Túnica Conjuntiva/efeitos dos fármacos , Dermatite Irritante/diagnóstico , Irritantes/administração & dosagem , Animais , Embrião de Galinha , Galinhas , Membrana Corioalantoide/imunologia , Túnica Conjuntiva/metabolismo , Túnica Conjuntiva/patologia , Qualidade de Produtos para o Consumidor/normas , Dermatite Irritante/etiologia , Detergentes/administração & dosagem , Detergentes/química , Relação Dose-Resposta a Droga , Humanos , Irritantes/toxicidade , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Testes de Irritação da Pele/métodos , Dodecilsulfato de Sódio/administração & dosagem , Lágrimas/metabolismo , Fatores de Tempo
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