RESUMO
BACKGROUND: Periodontal disease is a chronic inflammatory disease. Given its high prevalence, especially in aging population, the detailed mechanisms about pathogenesis of periodontal disease are important issues for study. Neutrophil firstly infiltrates to periodontal disease-associated pathogen loci and amplifies the inflammatory response for host defense. However, excessive neutrophil-secreted neutrophil elastase (NE) damages the affected gingival. In lung and esophageal epithelium, NE had been proved to upregulate several growth factors including placenta growth factor (PGF). PGF is an angiogenic factor with proinflammatory properties, which mediates the progression of inflammatory disease. Therefore, we hypothesize excessive NE upregulates PGF and participates in the pathogenesis and progression of periodontal disease. METHODS: In gingival epithelial cells (GEC), growth factors array demonstrated NE-increased growth factors and further be corroborated by Western blot assay and ELISA. The GEC inflammation was evaluated by ELISA. In mice, the immunohistochemistry results demonstrated ligature implantation-induced neutrophil infiltration and growth factor upregulation. By multiplex assay, the ligature-induced proinflammatory cytokines level in gingival crevicular fluid (GCF) were evaluated. Finally, alveolar bone absorption was analyzed by micro-CT images and H & E staining. RESULTS: NE upregulated PGF expression and secretion in GEC. PGF promoted GEC to secret IL-1ß, IL-6, and TNF-α in GCF In periodontal disease animal model, ligature implantation triggered NE infiltration and PGF expression. Blockade of PGF attenuated the ligature implantation-induced IL-1ß, IL-6, TNF-α and MIP-2 secretion and ameliorated the alveolar bone loss in mice. CONCLUSION: In conclusion, the NE-induced PGF triggers gingival epithelium inflammation and promotes the pathogenesis and progression of periodontal disease.
Assuntos
Gengivite , Doenças Periodontais , Animais , Camundongos , Indutores da Angiogênese/análise , Citocinas , Líquido do Sulco Gengival/química , Inflamação , Interleucina-6/análise , Elastase de Leucócito/análise , Fator de Crescimento Placentário/análise , Fator de Necrose Tumoral alfa/análiseRESUMO
Phospholipids not only have high nutritional value, but also have a positive effect on cardiovascular disease, cancer and nervous system diseases. However, the activity of individual phospholipid classes of shrimp phospholipids is rarely studied. This paper researched phospholipids in the by-products of Penaeus vannamei processing. The phospholipid classes of the head from P. vannamei (PV) were separated by column chromatography, analyzed with UHPLC-Q-Exactive HF/MS, and quantified using ammonium ferrothiocyarate spectrophometry. In addition, their cardiovascular activities in zebrafish models were evaluated. A total of 5 phospholipid classes were obtained, including PV-PC, PV-PE, PV-PI, PV-PS and PV-SM, and identified as phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS) and sphingomyelin (SM), respectively. In the phospholipid profiling analysis, PV-PC (308 molecules) had the highest proportion with 85.24%, followed by PV-PE (139 types) with 9.32%, PV-SM (41 structures) with 4.75%, PV-PS (24 types) with 0.16%, and PV-PI (6 molecules) with 0.03%. In the quantitative analysis, the content of PV was 45.7%, and the purity of phospholipid classes was 75.5-88.1%. In the cardiovascular activity assays, the effects of different phospholipid classes were different. For example, PV-PC groups had strong angiogenesis activity, but PV-PE groups showed the opposite property. Our comprehensive profiling analysis and in vivo bioactivity evaluation of phospholipids from the head of P. vannamei can provide evidence for their targeted applications in the future.
Assuntos
Fármacos Cardiovasculares , Penaeidae/química , Fosfolipídeos , Indutores da Angiogênese/análise , Indutores da Angiogênese/química , Indutores da Angiogênese/isolamento & purificação , Indutores da Angiogênese/farmacologia , Animais , Fármacos Cardiovasculares/análise , Fármacos Cardiovasculares/química , Fármacos Cardiovasculares/isolamento & purificação , Fármacos Cardiovasculares/farmacologia , Coração/efeitos dos fármacos , Fosfolipídeos/análise , Fosfolipídeos/química , Fosfolipídeos/isolamento & purificação , Fosfolipídeos/farmacologia , Peixe-ZebraRESUMO
OBJECTIVE: This study aimed to evaluate whether state and trait anxiety among pregnant women were associated with fetoplacental Doppler findings, abnormal placental pathology, and placental angiogenic factors. MATERIALS AND METHODS: A total of 102 pregnant women at 32-35 gestational weeks were recruited and examined prospectively. State and trait anxiety were measured using the State-Trait Anxiety Inventory. Using Doppler ultrasound, pulsatility index (PI) of the umbilical artery (UA), middle cerebral artery (MCA), and uterine artery (UtA) and cerebroplacental ratio (CPR) were determined. Doppler parameters were converted into multiples of the median (MoM). Abnormal placental pathology was classified into 2 groups: vascular underperfusion (VU) and histological chorioamnionitis (HCA). Immunohistochemical analysis was performed to examine placental cells staining positive for placental growth factor (PLGF) and hypoxia-inducible factor-1-α (HIF-1α), which are markers for angiogenesis and hypoxic status, respectively. RESULTS: Women with high state anxiety scores had low MCA-PI MoM and CPR MoM, while those with high trait anxiety scores had low MCA-PI MoM. VU was associated with a higher incidence of high trait anxiety scores, and HCA was associated with a higher incidence of high state and trait anxiety scores. Regression analysis showed a relationship between maternal state anxiety on MCA-PI MoM and HCA after controlling for covariates. Maternal trait anxiety exhibited relationships with VU and HCA after adjustment. CONCLUSION: Our results demonstrated that maternal anxiety is associated with altered fetal cerebral blood flow and abnormal placental pathology but is not associated with uteroplacental insufficiency and placental angiogenic factors.
Assuntos
Ansiedade/diagnóstico por imagem , Feto/irrigação sanguínea , Placenta/diagnóstico por imagem , Complicações na Gravidez/diagnóstico por imagem , Ultrassonografia Doppler , Ultrassonografia Pré-Natal/métodos , Adulto , Indutores da Angiogênese/análise , Ansiedade/patologia , Biomarcadores/análise , Circulação Cerebrovascular , Corioamnionite/diagnóstico por imagem , Corioamnionite/psicologia , Feminino , Hipóxia Fetal/diagnóstico por imagem , Hipóxia Fetal/embriologia , Hipóxia Fetal/psicologia , Feto/diagnóstico por imagem , Idade Gestacional , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/análise , Artéria Cerebral Média/diagnóstico por imagem , Placenta/patologia , Fator de Crescimento Placentário/análise , Gravidez , Complicações na Gravidez/patologia , Complicações na Gravidez/psicologia , Estudos Prospectivos , Fluxo Pulsátil , Artérias Umbilicais/diagnóstico por imagem , Artéria Uterina/diagnóstico por imagemRESUMO
Dragon blood has been used in wound treatment for many years and can be obtained from several distinct plant species. Dracorhodin, the active substituent of dragon blood, is a characteristic compound of the palm tree, Daemonorops draco. At present, the only method to evaluate the quality of commercial dragon blood samples is a HPLC method which determines the amount of dracorhodin in a dragon blood sample. In this study, we used zebrafish embryos as a platform to demonstrate the in vivo pro-angiogenic activity of dracorhodin perchlorate, the chemically synthesized analog of dracorhodin. By using this platform, three different commercial dragon blood samples were also examined. Our results clearly show that even though the commercial dragon blood samples had similar amounts of dracorhodin, they showed highly variable biological activity, such as pro-angiogenic effects and toxicity. In short, an in vivo activity assay platform for rapidly examining the biological activity of commercial dragon blood samples was successfully established here, which complements the current HPLC-based assay method.
Assuntos
Indutores da Angiogênese/farmacologia , Benzopiranos/farmacologia , Avaliação Pré-Clínica de Medicamentos/métodos , Extratos Vegetais/farmacologia , Indutores da Angiogênese/análise , Animais , Benzopiranos/análise , Linhagem Celular Tumoral , Cromatografia Líquida de Alta Pressão , Extratos Vegetais/análise , Peixe-Zebra/embriologiaRESUMO
Evaluation of cytokines in patients with diabetic retinopathy (DR) is important for the identification of future additive or alternative treatment options. Therefore, vitreous samples were obtained from patients with DR and patients with macular hole or macular pucker (control group) during 23-gauge-vitrectomy (n = 17/group). The levels of three pro-inflammatory (IL-1ß, IL-6, IFN-γ) and pleiotropic cytokines (IL-2, IL-4, IL-13) as well as VEGF, VEGF-A, and PGF were measured using an enzyme linked immunosorbent assay (ELISA). IL-1ß (p = 0.02) and IFN-γ (p = 0.04), two of the three tested pro-inflammatory cytokines, were elevated in the DR patients, while IL-6 (p = 0.51) level was comparable in both groups. Moreover, in DR samples, a trend towards an IL-13 down-regulation (p = 0.36) was observable. The IL-2 (p = 0.62) and IL-4 (p = 0.78) levels were comparable in both groups. All analyzed angiogenetic factors were up-regulated in DR patients (VEGF: p<0.001; VEGF-A: p = 0.002; PGF: p<0.001). The up-regulation of angiogenetic factors underline their importance in DR development. However, the interaction of the other cytokines showed an interesting pattern. Pro-inflammatory cytokines were also up-regulated, which could be evidence for inflammation processes in the diabetic retina. Furthermore, it seems that a counter response of immunomodulatory cytokines is in an initial process, but not strong enough to regulate the processes. Therefore, to support these anti-inflammatory mechanisms might be additive treatment option in the future.
Assuntos
Indutores da Angiogênese/análise , Citocinas/análise , Retinopatia Diabética/patologia , Corpo Vítreo/metabolismo , Idoso , Idoso de 80 Anos ou mais , Indutores da Angiogênese/metabolismo , Estudos de Casos e Controles , Citocinas/metabolismo , Retinopatia Diabética/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Interferon gama/análise , Interferon gama/metabolismo , Interleucina-1beta/análise , Interleucina-1beta/metabolismo , Interleucina-6/análise , Interleucina-6/metabolismo , Masculino , Pessoa de Meia-Idade , Fator de Crescimento Placentário/análise , Retina/metabolismo , Regulação para Cima , Fator A de Crescimento do Endotélio Vascular/análise , Fator A de Crescimento do Endotélio Vascular/metabolismo , VitrectomiaRESUMO
OBJECTIVE: Bone marrow cell extract (BMCE) was previously reported to restore salivary gland hypofunction caused by irradiation injury. Proteins were shown to be the main active factors in BMCE. However, BMCE therapy requires multiple injections and protein denaturation is a concern during BMCE storage. This study aimed to preserve, by lyophilization (freeze-drying), the bioactive factors in BMCE. METHODS: We developed a method to freeze-dry BMCE and then to analyze its ingredients and functions in vivo. Freeze-dried (FD) BMCE, freshly prepared BMCE (positive control), or saline (vehicle control) was injected into the tail vein of mice that had received irradiation to damage their salivary glands. RESULTS: Results demonstrated that the presence of angiogenesis-related factors and cytokines in FD-BMCE remained comparable to those found in fresh BMCE. Both fresh and FD-BMCE restored comparably saliva secretion, increased cell proliferation, upregulated regenerative/repair genes, protected salivary acinar cells, parasympathetic nerves, and blood vessels from irradiation-damaged salivary glands. CONCLUSION: Lyophilization of BMCE maintained its bioactivity and therapeutic effect on irradiation-injured salivary glands. The advantages of freeze-drying BMCE are its storage and transport at ambient temperature.
Assuntos
Células da Medula Óssea , Extratos Celulares/farmacologia , Lesões Experimentais por Radiação/tratamento farmacológico , Glândulas Salivares/fisiologia , Salivação/efeitos dos fármacos , Células Acinares/fisiologia , Indutores da Angiogênese/análise , Animais , Extratos Celulares/química , Proliferação de Células/efeitos dos fármacos , Citocinas/análise , Feminino , Liofilização , Camundongos , Neovascularização Fisiológica/efeitos dos fármacos , Glândulas Salivares/citologiaRESUMO
Plexiform lesions are characteristic histological changes of pulmonary arteries in human patients with severe pulmonary arterial hypertension (PAH) and are regarded as angiogenic lesions. Meat-type broiler chickens are susceptible to PAH and can develop plexiform lesions spontaneously. Whether the lesion development in broilers is associated with PAH predisposition and lung angiogenic environment remains unclear. Moreover, little is known about the cellular origin of these structures. In this work, plexiform lesions were detected in both layer chickens (a strain known to be resistant to PAH) and broiler chickens aged between 1 and 6 wk with normal pulmonary arterial pressures. Within each of the sampled ages, the lesion density did not differ between strains, with an exception of wk 4 when broiler was higher than layer. In contrast to the trend of age-related decline in layers, lesion densities in broilers demonstrated bi-phasic alterations characterized by a gradual decrease during wk 1 to 3 followed by a sudden increase at wk 4. The mRNA of 6 angiogenic factors in the lung tissue, namely, vascular endothelial growth factor receptor (VEGFR)-2, angiopoietin (Ang)-1, angiopoietin receptor Tie-2, transforming growth factor (TGF)-ß1, hepatocyte growth factor (HGF), and interleukin (IL)-8, were differentially expressed between strains. However, none of them was found to be significantly correlated with the lesion density by strain and age-adjusted partial correlation analysis. An in vivo experiment revealed impaired differentiation of endothelial progenitor cells (EPC) into endothelial cells during the producing of plexiform lesions, as evidenced by increased expression of endothelial CD133, a maker of EPC, but reduced expression of CD31, a marker of mature endothelial cells, in the parent vessels of plexiform lesions compared to normal vessels. Collectively, it appears unlikely that the predisposition to PAH or intrapulmonary angiogenic environment contributes to the lesion development in broilers when compared with layers. It is suggested that the lesion development is associated with increased pulmonary arterial pressure, and that local EPC dysfunction may play a role in the process.
Assuntos
Proteínas Aviárias/genética , Galinhas , Pneumopatias/veterinária , Doenças das Aves Domésticas/genética , Doenças das Aves Domésticas/patologia , Indutores da Angiogênese/análise , Animais , Proteínas Aviárias/metabolismo , Células Progenitoras Endoteliais/fisiologia , Feminino , Perfilação da Expressão Gênica/veterinária , Pneumopatias/genética , Pneumopatias/patologiaRESUMO
This study aimed to evaluate the effects of thyroid hormone on the decidua and metrial gland of rats and to examine the expression of angiogenic factors. 72 adult, female rats were divided into hypothyroid, T4-treated2, and control groups. At 10, 14 and 19 days of gestation (DG), the decidua and metrial gland were collected for histomorphometric and immunohistochemical evaluation of the expression of VEGF, Flk-1 and Tie-2. Hypothyroidism reduced the area of the decidua at 10 and 19 DG. Furthermore, VEGF was increased at 10 and 14 DG, and Flk-1 only at 14 DG, but both was reduced at 19 DG in the metrial gland without significantly changing the area occupied by blood vessels. Rats treated with T4 showed an increase in the decidua blood vessels at 10 and 19 DG. However, at 10 DG, excess T4 resulted in increased of Flk-1 in the decidua and metrial gland. Hypothyroidism increased the Tie-2 at 10 and 19 DG in the decidua and metrial gland. In conclusion, hypothyroidism reduces the area of the decidua and increases the expression of VEGF, Tie-2 and Flk-1. The excess of T4 promotes tissue angiogenesis by increasing the number of vessels in the decidua because of the increased expression of Flk-1.(AU)
Este estudo teve como objetivo avaliar os efeitos dos hormônios tireoidianos sobre a decídua e a glândula metrial pela análise da expressão de fatores angiogênicos em ratas. 72 ratas adultas, fêmeas foram distribuídas nos grupos hipotiroideo, tratado com T4 e controle. Aos 10, 14 e 19 dias de gestação (DG), a decídua e a glândula metrial foram coletadas para avaliação histomorfométrica e imunoistoquímica da expressão de VEGF, Flk-1 e Tie-2. O hipotireoidismo reduziu a área da decídua aos 10 e 19 DG. Além disso, o VEGF aumentou aos 10 e 14 DG e o Flk-1 apenas aos 14 DG, mas ambos foram reduzidos aos 19 DG na glândula metrial sem alterar significativamente a área ocupada pelos vasos sanguíneos. As ratas tratadas com T4 apresentaram aumento do número de vasos sanguíneos na decídua aos 10 e 19 DG. Além disso, aos 10 DG, o excesso de T4 resultou no aumento de Flk-1 na decídua e na glândula metrial. O hipotireoidismo aumentou o Tie-2 em 10 e 19 DG na decídua e na glândula metrial. Desta forma, pode-se concluir que o hipotireoidismo reduz a área da decídua e aumenta a expressão de VEGF, Tie-2 e Flk-1. O excesso de T4 promove a angiogênese tecidual ao aumentar o número de vasos na decídua devido ao aumento da expressão de Flk-1.(AU)
Assuntos
Animais , Feminino , Ratos , Feniltioureia/análise , Hormônios Tireóideos/análise , Decídua , Indutores da Angiogênese/análise , Glândula MetrialRESUMO
This study aimed to evaluate Himatanthus drasticus latex in a mice wound healing experimental model. Animals were divided into four groups (n=7) according to the treatments: GI - saline 0.9% (control), GII - mineral oil (vehicle), GIII - H. drasticus commercial latex (HdCL) and GIV - H. drasticus mixed isolated fraction (MIF, 1 mg/mL). The treatments were applied topically once daily, 50 µL for 14 consecutive days. Macroscopic lesions were evaluated, considering parameters such as swelling, redness, granulation tissue and reepithelialization. VEGF+, CD68+ expressions and mast cells (Toluidin blue stain) were evaluated. HdCL induced higher contraction and exuberant granulation tissue (P > 0.05). HdCL showed a mild inflammatory process while MIF induced intense infiltrate inflammatory predominantly by lymphocytes, vascular congestion, bleeding and did not presented full reepithelialization. Reorganization of collagen fibers (red picrosirius stain) was observed. CD68+ expression and mast cells were presented as moderate, intense and mild in GI, GIII and GIV, respectively. Neovascularization occurred in all groups, while VEGF+ expression was intense in MIF in relation to HdCL. We concluded that HdCL presents wound healing potential, through modulation of mast cells, CD68+ and VEGF+ expressions that can be associated to triterpenes presence according MIF isolated from HdCL.(AU)
Objetivou-se avaliar o látex de Himatanthus drasticus em feridas induzidas experimentalmente em camundongos. Os animais foram divididos em quatro grupos (n=7): GI - salina 0,9% (controle), GII - óleo mineral (veículo), GIII - látex comercial de H. drasticus (HdCL) e GIV - fração isolada mista de H. drasticus (MIF, 1mg/mL). Os tratamentos foram aplicados topicamente uma vez ao dia (50µL), durante 14 dias consecutivos. Lesões macroscópicas, as expressões de VEGF+, CD68+ e a participação dos mastócitos (coloração azul de toluidina) foram avaliadas. HdCL induziu maior contração e tecido de granulação exuberante (P >0,05). HdCL induziu leve processo inflamatório enquanto MIF promoveu intenso infiltrado inflamatório predominantemente linfocítico, congestão vascular, hemorragia e reepitelização parcial. Observou-se reorganização das fibras colágenas (coloração picrosírius). A expressão de CD68+ e os mastócitos apresentaram-se moderados, intensos e leves em GI, GIII e GIV, respectivamente. A neovascularização foi observada em todos os grupos, enquanto a expressão de VEGF+ foi mais intensa em MIF em relação a HdCL. Conclui-se que HdCL apresenta potencial de cicatrização por meio da modulação dos mastócitos e das expressões de CD68+ e VEGF+, o que pode estar associado à presença de triterpenos de acordo com MIF isolada de HdCL.(AU)
Assuntos
Animais , Camundongos , Indutores da Angiogênese/análise , Apocynaceae/química , Glicoproteínas , Mastócitos , Fator A de Crescimento do Endotélio Vascular/análise , Cicatrização/efeitos dos fármacos , Látex/químicaRESUMO
OBJECTIVE: We investigated whether the level of vascular endothelial growth factor (VEGF), placental growth factor (PlGF), and soluble VEGF receptor-1 (sFlt-1) in midtrimester amniotic fluid of preterm birth have different values compared with term delivery. MATERIALS AND METHODS: Our participants were 86 pregnant women who had undergone amniocentesis from 16 to 19 weeks of gestation. Forty-three cases were women with preterm delivery, and the other 43 cases were matched women with full-term delivery. Stored amniotic fluid was investigated after the delivery. The levels of VEGF, PlGF, and sFlt-1 were measured by enzyme-linked immunosorbent assay and Western blot. RESULTS: The levels of VEGF and PlGF in the preterm group were significantly higher than in the control group (30.48 ± 8.57 pg/mL vs. 26.06 ± 8.24 pg/mL and 28.83 ± 7.83 pg/mL vs. 25.35 ± 8.26 pg/mL, respectively) (p = 0.017 and 0.048, respectively). In terms of sFlt-1, the levels were decreased in the preterm group (10,478.51 ± 4012.56 pg/mL vs. 12,544.05 ± 4140.96 pg/mL) (p = 0.021). CONCLUSION: This study explains that elevated levels of VEGF and PlGF, suggestive of angiogenesis and tendency of inflammation at midtrimester, are predictive of preterm delivery, and their availability is maximized by downregulation of sFlt-1.
Assuntos
Líquido Amniótico/química , Indutores da Angiogênese/análise , Segundo Trimestre da Gravidez , Nascimento Prematuro/etiologia , Adulto , Amniocentese , Biomarcadores/análise , Estudos de Casos e Controles , Feminino , Idade Gestacional , Humanos , Fator de Crescimento Placentário/análise , Valor Preditivo dos Testes , Gravidez , Estudos Prospectivos , Fatores de Risco , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/análise , Fatores de Crescimento do Endotélio Vascular/análiseRESUMO
The T-helper cell type 2-promoting cytokine interleukin-33 (IL-33) has been implicated in asthma pathogenesis. Angiogenesis is a feature of airways remodelling in asthma. We hypothesized that IL-33 induces airways angiogenesis and expression of angiogenic factors in an established murine surrogate of asthma. In the present study, BALB/c mice were subjected to serial intranasal challenge with IL-33 alone for up to 70 days. In parallel, ovalbumin (OVA) -sensitized mice were subjected to serial intranasal challenge with OVA or normal saline to serve as positive and negative controls, respectively. Immunohistochemical analysis of expression of von Willebrand factor and erythroblast transformation-specific-related gene, both blood vessel markers, and angiogenic factors angiogenin, insulin-like growth factor-1, endothelin-1, epidermal growth factor and amphiregulin was performed in lung sections ex vivo. An established in-house assay was used to test whether IL-33 was able to induce microvessel formation by human vascular endothelial cells. Results showed that serial intranasal challenge of mice with IL-33 or OVA resulted in proliferation of peribronchial von Willebrand factor-positive blood vessels to a degree closely related to the total expression of the angiogenic factors amphiregulin, angiogenin, endothelin-1, epidermal growth factor and insulin-like growth factor-1. IL-33 also induced microvessel formation by human endothelial cells in a concentration-dependent fashion in vitro. Our data are consistent with the hypothesis that IL-33 has the capacity to induce angiogenesis at least partly by increasing local expression of multiple angiogenic factors in an allergen-independent murine asthma surrogate, and consequently that IL-33 or its receptor is a potential novel molecular target for asthma therapy.
Assuntos
Indutores da Angiogênese/análise , Asma/tratamento farmacológico , Interleucina-33/administração & dosagem , Pulmão/irrigação sanguínea , Neovascularização Fisiológica/efeitos dos fármacos , Administração Intranasal , Animais , Asma/fisiopatologia , Fator de Crescimento Epidérmico/análise , Feminino , Interleucina-33/farmacologia , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Artificial generated buccal mucosa equivalents are a promising approach for the reconstruction of urethral defects. Limiting in this approach is a poor blood vessel supply after transplantation, resulting in increased morbidity and necrosis. We generated a pre-vascularized buccal mucosa equivalent in a tri-culture of primary buccal epithelial cells, fibroblasts and microvascular endothelial cells, using a native collagen membrane as a scaffold. A successful pre-vascularization and dense formation of capillary-like structures at superficial areas was demonstrated. The lumen size of pre-formed blood vessels corresponded to the capillary size in vivo (10-30 µm). Comparing native with a highly cross-linked collagen membrane we found a distinct higher formation of capillary-like structures on the native membrane, apparently caused by higher secretion of angiogenic factors such as PDGF, IL-8 and angiopoietin by the cells. These capillary-like structures became functional blood vessels through anastomosis with the host vasculature after implantation in nude mice. This in vitro method should result in an accelerated blood supply to the biomaterial with cells after transplantation and increase the succes rates of the implant material.
Assuntos
Células Endoteliais/citologia , Células Epiteliais/citologia , Fibroblastos/citologia , Mucosa Bucal , Organoides/irrigação sanguínea , Engenharia Tecidual/métodos , Transplantes/irrigação sanguínea , Indutores da Angiogênese/análise , Animais , Capilares/citologia , Capilares/crescimento & desenvolvimento , Células Cultivadas , Técnicas de Cocultura , Colágeno , Prepúcio do Pênis/citologia , Gengiva/citologia , Xenoenxertos , Humanos , Masculino , Membranas Artificiais , Camundongos , Camundongos Nus , Organoides/citologia , Molécula-1 de Adesão Celular Endotelial a Plaquetas/análise , Alicerces TeciduaisRESUMO
Ticks are blood-feeding arthropods with an outstanding ability to remain attached to its host for considerable periods while blood-feeding and remaining unnoticed. Their success results from the ability to modulate hemostatic and host immune responses. The ability to "bypass" a host's defenses, prevent blood clotting and wound healing makes ticks utterly interesting animals for the development of new drugs. Studies worldwide on various tick species have shown that tick saliva possesses a wide array of lipidic and proteic biomolecules with useful properties. These include not only immunomodulatory, anti-inflammatory, anti-platelet and anti-clotting properties, but also cytotoxic and cytolitic properties that act against various cell types, and anti-angiogenic properties, which have gained increasing prominence. We searched PubMed, Science Direct, Elsevier and other sites for publications regarding tick saliva and its effects on cancer cells and angiogenesis. Our aim was to compile a list of molecules with potential for host adaptation and for the development of new cancer treatment drugs.
Assuntos
Indutores da Angiogênese/análise , Indutores da Angiogênese/farmacocinética , Antineoplásicos/análise , Antineoplásicos/farmacocinética , Neoplasias/tratamento farmacológico , Saliva/química , Carrapatos/química , Animais , HumanosRESUMO
BACKGROUND: A fundamental issue limiting the efficacy of surgical approaches designed to correct periodontal mucogingival defects is that new tissues rely on limited sources of blood supply from the adjacent recipient bed. Accordingly, therapies based on tissue engineering that leverage local self-healing potential may represent promising alternatives for the treatment of mucogingival defects by inducing local vascularization. The aim of this study is to evaluate the effect of commercially available living cellular sheets (LCS) on the angiogenic potential of neonatal dermal human microvascular endothelial cells (HMVEC-dNeo). METHODS: The effect of LCS on HMVEC-dNeo proliferation, migration, capillary tube formation, gene expression, and production of angiogenic factors was evaluated over time. RESULTS: LCS positively influenced HMVEC-dNeo proliferation and migration. Moreover, HMVEC-dNeo incubated with LCS showed transcriptional profiles different from those of untreated cells. Whereas increased expression of angiogenic genes predominated early on in response to LCS, late-phase responses were characterized by up- and downregulation of angiostatic and angiogenic genes. However, this trend was not confirmed at the protein level, as LCS induced increased production of most of the angiogenic factors tested (i.e., epidermal growth factor [EGF], heparin-binding EGF-like growth factor, interleukin 6, angiopoietin, platelet-derived growth factor-BB, placental growth factor, and vascular endothelial growth factor) throughout the investigational period. Finally, although LCS induced HMVEC-dNeo proliferation, migration, and expression of angiogenic factors, additional factors and environmental pressures are likely to be required to promote the development of complex, mesh-like vascular structures. CONCLUSION: LCS favor initial mechanisms that govern angiogenesis but failed to enhance or accelerate HMVEC-dNeo morphologic transition to complex vascular structures.
Assuntos
Células Endoteliais/fisiologia , Endotélio Vascular/citologia , Microvasos/citologia , Neovascularização Fisiológica/fisiologia , Alicerces Teciduais , Indutores da Angiogênese/análise , Angiopoietinas/análise , Becaplermina , Capilares/fisiologia , Movimento Celular/fisiologia , Proliferação de Células , Sobrevivência Celular/fisiologia , Colágeno Tipo I/química , Fator de Crescimento Epidérmico/análise , Fibroblastos/fisiologia , Regulação da Expressão Gênica/genética , Fator de Crescimento Semelhante a EGF de Ligação à Heparina/análise , Humanos , Interleucina-6/análise , Queratinócitos/fisiologia , Neovascularização Fisiológica/genética , Fator de Crescimento Placentário , Proteínas da Gravidez/análise , Proteínas Proto-Oncogênicas c-sis/análise , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Fator A de Crescimento do Endotélio Vascular/análiseRESUMO
BACKGROUND: Advanced glycation end products (AGE) have been found in inflamed gingival tissue and have been shown to interfere with the integrity of extracellular matrix and cell-matrix interactions. This study aims to investigate the modulatory effect of aminoguanidine (AG), an AGE inhibitor, in various stages of experimental periodontitis. METHODS: Thirty-six Sprague-Dawley rats were used. AG or normal saline (NS) was systemically administered in the induction, progression, and recovery phases of ligature-induced periodontitis. Dynamic changes of the periodontium were evaluated by microcomputed tomography, histology, and immunohistochemistry of the receptor for AGE (RAGE). Molecular mechanisms were evaluated by myeloperoxidase activity, gene expression of RAGE, and markers associated with tissue repair and homeostasis, including vascular endothelial growth factor (VEGF), type I collagen, fibronectin, and periostin. RESULTS: AG appeared to inhibit the degradation of the collagen matrix in the induction phase but promoted collagen reorganization in the progression and recovery phases of experimental periodontitis. In the induction sites, periodontal bone loss was significantly reduced (P <0.05), with significantly reduced RAGE (P <0.05) and significantly elevated fibronectin and periostin levels (P <0.01). No significant alterations in the levels of myeloperoxidase, VEGF, and collagen were noted. In the progression and recovery sites, similar trends were observed, with insignificant differences relative to NS-treated animals. CONCLUSIONS: AG reduced periodontal bone loss during the induction of experimental periodontitis, and the effects appeared to be insignificant in the progression and recovery phases. This modulation was related to the inhibition of the AGE-RAGE axis to resume cell-matrix interactions and maintain tissue integrity.
Assuntos
Produtos Finais de Glicação Avançada/efeitos dos fármacos , Guanidinas/uso terapêutico , Óxido Nítrico Sintase/antagonistas & inibidores , Periodontite/prevenção & controle , Receptores Imunológicos/efeitos dos fármacos , Perda do Osso Alveolar/patologia , Perda do Osso Alveolar/fisiopatologia , Perda do Osso Alveolar/prevenção & controle , Processo Alveolar/efeitos dos fármacos , Processo Alveolar/patologia , Indutores da Angiogênese/análise , Animais , Biomarcadores/análise , Moléculas de Adesão Celular/efeitos dos fármacos , Colágeno Tipo I/efeitos dos fármacos , Modelos Animais de Doenças , Progressão da Doença , Fibronectinas/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Processamento de Imagem Assistida por Computador/métodos , Masculino , Periodontite/patologia , Periodontite/fisiopatologia , Periodonto/efeitos dos fármacos , Periodonto/patologia , Peroxidase/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Receptor para Produtos Finais de Glicação Avançada , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Cicatrização/fisiologia , Microtomografia por Raio-X/métodosRESUMO
BACKGROUND: To investigate whether the immunohistochemical expression of GLUT-1 and HIF-1α is related to the diagnosis and pathogenesis of oral vascular lesions. STUDY METHODS: Thirty cases each of pyogenic granuloma (PG) and hemangioma were studied. Antibodies against GLUT-1 and HIF-1α were detected by immunoperoxidase staining in 3-µm histological sections, and the results were analyzed qualitatively and quantitatively, respectively. Positive and negative cells were counted, and the mean number of positive cells was calculated for each case. RESULTS: The initial diagnosis of hemangioma was maintained in only 7 (23%) of the 30 cases studied, which were positive for GLUT-1. The remaining 23 cases were reclassified as vascular malformation (VM) (n = 13) and PG (n = 10) due to the absence of staining. The endothelium of blood vessels was negative for GLUT-1 in all cases initially diagnosed as PG (n = 30). The percentage of HIF-1α-positive cells was higher in cases of PG, followed by hemangiomas and VMs (P = 0.005). CONCLUSIONS: Histological features are not sufficient to establish the correct diagnosis of oral hemangiomas, and an accurate anamnesis is essential in these cases. In addition, these findings demonstrate that vascular lesions express mediators of angiogenesis, HIF-1α, and suggest that his process may play a role in the pathogenesis of vascular.
Assuntos
Transportador de Glucose Tipo 1/análise , Granuloma Piogênico/diagnóstico , Hemangioma/diagnóstico , Subunidade alfa do Fator 1 Induzível por Hipóxia/análise , Doenças da Boca/diagnóstico , Neoplasias Bucais/diagnóstico , Adulto , Indutores da Angiogênese/análise , Biomarcadores/análise , Biomarcadores Tumorais/análise , Diagnóstico Diferencial , Endotélio Vascular/patologia , Granuloma Piogênico/etiologia , Granuloma Piogênico/patologia , Hemangioma/etiologia , Hemangioma/patologia , Humanos , Técnicas Imunoenzimáticas , Doenças da Boca/etiologia , Doenças da Boca/patologia , Neoplasias Bucais/etiologia , Neoplasias Bucais/patologia , Estudos Retrospectivos , Malformações Vasculares/diagnóstico , Malformações Vasculares/etiologia , Malformações Vasculares/patologiaRESUMO
PURPOSE: Oral pregnancy tumors (OPTs) arise on the inflamed gingiva of women after the first trimester of pregnancy. The expression of angiogenic markers and female hormone receptors was assessed. MATERIALS AND METHODS: Immunohistochemistry was used to analyze the expression of estrogen and progesterone receptors and the expression of angiogenic factors, such as vascular endothelial growth factor (VEGF) and its receptor, fibroblast growth factor (FGF), and hypoxia inducible factors 1α and 3α (HIF1α and HIF3α). Experimental groups included 9 OPTs, 10 oral pyogenic granulomas from nonpregnant women of the same age, and 9 oral pyogenic granulomas from postmenopausal women. RESULTS: VEGF expression in stromal histiocytes and endothelial cells of small vessels was positively correlated in the OPT group (P < .05 by χ(2) test). VEGF receptor also was overexpressed in stromal histiocytes and endothelial cells of OPTs compared with oral pyogenic granulomas from nonpregnant and postmenopausal women (P < .005 by χ(2) test). No correlation was detected among estrogen and progesterone receptors, FGF and HIF1α and HIF3α (ER and PgR respectively) in the 3 experimental groups. CONCLUSIONS: VEGF-associated angiogenesis is most likely involved in the pathogenesis of the lesion. These results imply that local inhibition of VEGF activity could be an adjuvant therapeutic approach for OPTs to control hemorrhage, which can be massive at the surgical excision of such lesions during pregnancy.
Assuntos
Indutores da Angiogênese/análise , Neoplasias Gengivais/metabolismo , Fator 1 Induzível por Hipóxia/análise , Neovascularização Patológica/metabolismo , Complicações Neoplásicas na Gravidez/metabolismo , Receptores de Progesterona/biossíntese , Receptores de Fatores de Crescimento do Endotélio Vascular/biossíntese , Fator A de Crescimento do Endotélio Vascular/biossíntese , Adulto , Estudos de Casos e Controles , Distribuição de Qui-Quadrado , Feminino , Fatores de Crescimento de Fibroblastos/análise , Neoplasias Gengivais/complicações , Granuloma Piogênico/metabolismo , Humanos , Pessoa de Meia-Idade , Neovascularização Patológica/complicações , Pós-Menopausa , Gravidez , Receptores de Estrogênio/biossíntese , Adulto JovemRESUMO
BACKGROUND: Angiogenin (ANG) is a member of the ribonuclease superfamily and of medical interest largely because it supports the growth of primary and metastatic malignancies. This study is the first to investigate the potential role of ANG in tongue carcinoma neo-angiogenesis and cancer cell proliferation. METHODS: Angiogenin expression (in carcinoma cells and endothelial intratumor vessel cells), CD105-assessed micro-vessel density (MVD), and MIB-1 expression were correlated with prognostic parameters in 28 primarily consecutively operated pT1-T2 tongue carcinomas (squamous cell carcinoma [SCC]). Whenever feasible, a computer-based image analysis system was used for the immunohistochemical reaction analysis. RESULTS: No significant correlations emerged between ANG expression in the tongue carcinoma cells or endothelial intratumor vessel cells and tongue SCC recurrence rate or disease-free survival (DFS). ANG expression was also unrelated to CD105-assessed MVD or MIB-1 expression. Conversely, CD105-assessed MVD correlated directly with recurrence rate (P = 0.02) and DFS was significantly shorter in cases with CD105-assessed MVD >167 micro-vessels/mm(2) than in those with CD105-assessed MVD ≤167 micro-vessels/mm(2) (P = 0.042). CONCLUSIONS: Our results support the hypothesis that CD105-assessed MVD would be a valuable parameter for predicting which patients with tongue SCC are at greatest risk of disease recurrence. Despite our study results, the role of ANG in tongue carcinoma warrants further investigation in larger series.
Assuntos
Indutores da Angiogênese/farmacologia , Carcinoma de Células Escamosas/irrigação sanguínea , Neovascularização Patológica/patologia , Ribonuclease Pancreático/fisiologia , Neoplasias da Língua/irrigação sanguínea , Indutores da Angiogênese/análise , Antígenos CD/análise , Proliferação de Células/efeitos dos fármacos , Intervalo Livre de Doença , Endoglina , Células Endoteliais/patologia , Endotélio Vascular/patologia , Seguimentos , Humanos , Processamento de Imagem Assistida por Computador/métodos , Antígeno Ki-67/análise , Microvasos/patologia , Recidiva Local de Neoplasia/patologia , Estadiamento de Neoplasias , Prognóstico , Receptores de Superfície Celular/análise , Ribonuclease Pancreático/análise , Resultado do TratamentoRESUMO
BACKGROUND: Interleukin 8 (IL-8) is a pro-angiogenic, pro-inflammatory mediator that belongs to the family of chemokines. Due to its pro-angiogenic characteristic, it may play a vital role in tumour angiogenesis and progression. OBJECTIVES: This study was designed to estimate the levels of salivary IL-8 in oral precancer and oral squamous cell carcinoma (OSCC) patients and compare them with healthy controls. The aim was to evaluate its efficacy as a potential biomarker for these diseases. MATERIALS AND METHODS: Each group comprised 25 individuals. The salivary IL-8 levels were determined by enzyme-linked immunosorbent assay. RESULTS: The levels of salivary IL-8 were found to be significantly elevated in patients with OSCC as compared to the precancer group (p < 0.0001) and healthy controls (p < 0.0001). However, the difference in salivary IL-8 concentrations among the precancer group and controls was statistically non-significant (p = 0.738). CONCLUSIONS: Our results suggested that salivary IL-8 can be utilised as a potential biomarker for OSCC. Salivary IL-8 was found to be non-conclusive for oral premalignancy in this preliminary study. Hence, its possible role in transition from premalignancy to malignancy needs further research with larger sample sizes. CLINICAL RELEVANCE: Saliva as a diagnostic biofluid offers a number of advantages over blood-based testing. The role of IL-8 in oral cancer if validated further by future research can provide an easy diagnostic test as well as a prognostic indicator for patients undergoing treatment. Therefore, if it's role in tumourigenesis can be sufficiently assessed, it could open up new avenues to find out novel treatment modalities for oral cancer.
Assuntos
Carcinoma de Células Escamosas/imunologia , Interleucina-8/análise , Neoplasias Bucais/imunologia , Lesões Pré-Cancerosas/imunologia , Saliva/imunologia , Proteínas e Peptídeos Salivares/análise , Adolescente , Adulto , Idoso , Indutores da Angiogênese/análise , Areca , Biomarcadores Tumorais/análise , Estudos de Casos e Controles , Feminino , Humanos , Mediadores da Inflamação/análise , Leucoplasia Oral/imunologia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Fibrose Oral Submucosa/imunologia , Fumar , Uso de Tabaco , Adulto JovemRESUMO
We determined the levels of the endogenous angiogenesis inhibitors soluble vascular endothelial growth factor receptor-1 (sVEGFR-1), thrombospondin (TSP)-1 and TSP-2 in the vitreous fluid from patients with proliferative diabetic retinopathy (PDR) and correlated their levels with clinical disease activity and the levels of vascular endothelial growth factor (VEGF). Vitreous samples from 30 PDR and 25 nondiabetic patients were studied by enzyme-linked immunosorbent assay. TSP-1 was not detected. VEGF and TSP-2 levels were significantly higher in PDR with active neovascularization compared with inactive PDR and nondiabetic patients (P < 0.001 for both comparisons). VEGF, sVEGFR-1 and TSP-2 levels were significantly higher in PDR with hemorrhage compared with PDR without hemorrhage and nondiabetic patients (P = 0.0063; 0.0144; <0.001, respectively). VEGF and sVEGFR-1 levels were significantly higher in PDR without traction retinal detachment (TRD) compared with PDR with TRD and nondiabetic patients (P = 0.038; 0.022, respectively). TSP-2 levels were significantly higher in PDR with TRD compared with PDR without TRD and nondiabetic patients (P < 0.001). There was a significant correlation between levels of VEGF and sVEGFR-1 (r = 0.427, P = 0.038). Our findings suggest that upregulation of sVEGFR-1 and TSP-2 may be a protective mechanism against progression of angiogenesis associated with PDR. TSP-2 might be associated with TRD.