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1.
J Appl Oral Sci ; 28: e20190215, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31939521

RESUMO

OBJECTIVE: This study evaluated the angiogenesis-enhancing potential of a tricalcium silicate-based mineral trioxide aggregate (ProRoot MTA), Biodentine, and a novel bioceramic root canal sealer (Well-Root ST) in human dental pulp stem cells (hDPSCs), human periodontal ligament stem cells (hPLSCs), and human tooth germ stem cells (hTGSCs). METHODOLOGY: Dulbecco's modified Eagle's medium was conditioned for 24 h by exposure to ProRoot MTA, Biodentine, or Well-Root ST specimens (prepared according to the manufacturers' instructions). The cells were cultured in these conditioned media and their viability was assessed with 3-(4,5-dimethyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H tetrazolium (MTS) on days 1, 3, 7, 10, and 14. Angiogenic growth factors [platelet-derived growth factor (PDGF), basic fibroblast growth factor (FGF-2), and vascular endothelial growth factor (VEGF)] were assayed by sandwich enzyme-linked immunosorbent assay (ELISA) on days 1, 7, and 14. Human umbilical vein endothelial cell (HUVEC) migration assays were used to evaluate the vascular effects of the tested materials at 6-8 h. Statistical analyses included Kruskal-Wallis, Mann-Whitney U, and Friedman and Wilcoxon signed rank tests. RESULTS: None of tricalcium silicate-based materials were cytotoxic and all induced a similar release of angiogenic growth factors (PDGF, FGF-2, and VEGF) (p>0.05). The best cell viability was observed for hDPSCs (p<0.05) with all tricalcium silicate-based materials at day 14. Tube formation by HUVECs showed a significant increase with all tested materials (p<0.05). CONCLUSION: The tricalcium silicate-based materials showed potential for angiogenic stimulation of all stem cell types and significantly enhanced tube formation by HUVECs.


Assuntos
Indutores da Angiogênese/farmacologia , Compostos de Cálcio/farmacologia , Cerâmica/farmacologia , Materiais Restauradores do Canal Radicular/farmacologia , Silicatos/farmacologia , Células-Tronco/efeitos dos fármacos , Materiais Biocompatíveis/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Fator 2 de Crescimento de Fibroblastos/análise , Fator 2 de Crescimento de Fibroblastos/efeitos dos fármacos , Citometria de Fluxo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Teste de Materiais , Neovascularização Fisiológica/efeitos dos fármacos , Ligamento Periodontal/citologia , Ligamento Periodontal/efeitos dos fármacos , Fator de Crescimento Derivado de Plaquetas/análise , Fator de Crescimento Derivado de Plaquetas/efeitos dos fármacos , Reprodutibilidade dos Testes , Estatísticas não Paramétricas , Germe de Dente/citologia , Germe de Dente/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/análise , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacos
2.
Support Care Cancer ; 28(1): 177-184, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31001696

RESUMO

PURPOSE: Crevicular fluid was used to assess interleukin-17 (IL-17) and vascular endothelial growth factor (VEGF) in cancer patients receiving zoledronic acid and/or bevacizumab. The markers were also assessed in the serum. METHODS: Twenty-five patients were included and comprised three groups: patients who received zoledronic acid (n = 9), patients who received bevacizumab (n = 9), and patients who received zoledronic acid combined with bevacizumab (n = 5). One patient received zoledronic acid and everolimus and another received zoledronic acid, bevacizumab, and temsirolimus. IL-17 and VEGF were measured by standard quantitative ELISA kits and assessed in two study points. RESULTS: Twenty-four patients maintained good periodontal health; one had asymptomatic osteonecrosis of the jaw. First assessment: 44 samples were collected; 21 from serum and 23 from crevicular fluid. Second assessment, 6 months later: 11 samples were collected; 6 from serum and 5 from crevicular fluid. IL-17 was detected in all samples, in serum and crevicular fluid, and remained unchanged at both time points. Serum VEGF in patients with bevacizumab alone or combined with zoledronic acid was significantly lower compared with that of patients who received zoledronic acid alone. VEGF was not detected in the crevicular fluid. CONCLUSIONS: Crevicular fluid might be an easy, non-invasive means to assess IL-17. The stable values of IL-17 in crevicular fluid and serum and the lack of VEGF in the crevicular fluid could be related to the good periodontal health of our patients. Further studies are needed to assess IL-17 and VEGF in the crevicular fluid in patients with and without periodontal disease.


Assuntos
Bevacizumab/administração & dosagem , Líquido do Sulco Gengival/química , Interleucina-17/análise , Neoplasias/tratamento farmacológico , Doenças Periodontais/diagnóstico , Fator A de Crescimento do Endotélio Vascular/análise , Ácido Zoledrônico/administração & dosagem , Idoso , Idoso de 80 Anos ou mais , Bevacizumab/efeitos adversos , Biomarcadores/análise , Biomarcadores/metabolismo , Quimioterapia Combinada/efeitos adversos , Feminino , Líquido do Sulco Gengival/metabolismo , Humanos , Inflamação/diagnóstico , Inflamação/metabolismo , Interleucina-17/metabolismo , Masculino , Pessoa de Meia-Idade , Neoplasias/irrigação sanguínea , Neoplasias/metabolismo , Neoplasias/patologia , Neovascularização Patológica/diagnóstico , Neovascularização Patológica/metabolismo , Osteonecrose/induzido quimicamente , Osteonecrose/diagnóstico , Osteonecrose/metabolismo , Doenças Periodontais/induzido quimicamente , Doenças Periodontais/etiologia , Bolsa Periodontal/induzido quimicamente , Bolsa Periodontal/diagnóstico , Bolsa Periodontal/metabolismo , Valor Preditivo dos Testes , Fator A de Crescimento do Endotélio Vascular/metabolismo , Ácido Zoledrônico/efeitos adversos
3.
BMC Complement Altern Med ; 19(1): 324, 2019 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-31752829

RESUMO

BACKGROUND: The present study was designed to investigate the effects of Berberis vulgaris (BV) juice consumption on plasma levels of insulin-like growth factor (IGF-1), IGF-binding proteins (IGFBPs), and the expression of PPAR-γ, VEGF and HIF in women with benign breast disease. METHODS: This parallel design randomized, double-blind controlled clinical trial was conducted on 85 eligible patients diagnosed with benign breast disease. They were assigned randomly into either BV juice group (n = 44, BV juice: 480 ml/day) or placebo group (n = 41, BV placebo juice: 480 ml/day) for 8 weeks intervention. Participants, caregivers and those who assessed laboratory analyses were blinded to the assignments. Plasma levels of biomarkers were measured at baseline and after 8 weeks by ELISA. Quantitative real-time PCR was used to measure the fold change in the expression of each interested gene. RESULTS: The compliance of participants was 95.2% and 40 available subjects analyzed in each group at last. Relative treatment (RT) effects for BV juice caused 16% fall in IGF-1 concentration and 37% reduction in the ratio of IGF-1/1GFBP1. Absolute treatment effect expressed 111 ng/ml increased mean differences of IGFBP-3 between BV group and placebo. Plasma level of PPAR-γ increased in both groups but it was not significant. Fold changes in the expressions of PPAR-γ, VEGF and HIF showed down-regulation in the intervention group compared to placebos (P < 0.05). CONCLUSIONS: The BV juice intervention over 8 weeks was accompanied by acceptable efficacy and decreased plasma IGF-1, and IGF-1/IGFBP-1 ratio partly could be assigned to enhanced IGFBP-1 level in women with BBD. The intervention caused reductions in the expression levels of PPAR, VEGF, and HIF which are remarkable genomic changes to potentially prevent breast tumorigenesis. TRIAL REGISTRATION: IRCT2012110511335N2. Registered 10 July 2013 (retrospectively registered).


Assuntos
Berberis , Neoplasias da Mama , Sucos de Frutas e Vegetais , Adulto , Mama/química , Mama/patologia , Neoplasias da Mama/química , Neoplasias da Mama/dietoterapia , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Feminino , Humanos , Fator 1 Induzível por Hipóxia/análise , Fator 1 Induzível por Hipóxia/metabolismo , Fator de Crescimento Insulin-Like I/análise , Fator de Crescimento Insulin-Like I/metabolismo , Pessoa de Meia-Idade , PPAR gama/análise , PPAR gama/metabolismo , Fator A de Crescimento do Endotélio Vascular/análise , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adulto Jovem
4.
Klin Lab Diagn ; 64(11): 663-668, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31747494

RESUMO

To study the concentration of vasculoendothelial growth factor (VEGF) in mixed saliva and serum of patients in normal conditions and with generalized periodontitis. The main group (n = 42) was represented by patients with generalized periodontitis. The comparison group (n = 36) consisted of patients without periodontal tissue diseases. The concentration of VEFR was determined by the method of enzyme-linked immunosorbent assay (ELISA) using a commercial test-system "VEGF - IFA - BEST" (A-8784) ("Vector - Best", Russia). The median values VEFR in saliva were 5.49 times higher than the values for serum in the main group (p = 0.000000) and 7.01 times in the comparison group (p = 0.000000). The concentration of VEFR in the saliva of the examined main group exceeded the similar values of the comparison group (p = 0,014857); the median and interquartile range for the main group was 1098.45 (925.5; 1291) pg/ml, and for the comparison group 1360.5 (998.9; 2062) pg/ml. There were no differences in the serum VEFR concentration (p = 0.775124). No significant correlation was found between the serum VEFR content and the mixed saliva. The Spearman's rank correlation coefficient for the main group was R = 0,0184358, and for the comparison group, respectively, R = 0.188932. The source of VEFR in saliva are the glands and cells of the oral mucosa, and not the process of exudation from blood serum. The high content of VEFR in the saliva of healthy people and a decrease in its level during periodontitis indicates the important role of this protein in the processes of maintaining the normal state of periodontal tissues and reparation of tissues of the oral mucosa.


Assuntos
Periodontite/diagnóstico , Saliva/química , Soro/química , Fator A de Crescimento do Endotélio Vascular/análise , Humanos , Federação Russa
5.
Anticancer Res ; 39(11): 6403-6412, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31704874

RESUMO

BACKGROUND: Cytokines, metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) take part in many processes involved in tumor progression and invasion such as degradation of the extracellular matrix, influence on immune cells associated with tumor tissue, and angiogenesis. Thus, the aim of this study was to compare the concentration of plasma levels and tissue expression of macrophage colony-stimulating factor (M-CSF), vascular endothelial growth factor (VEGF), matrix metalloproteinases (MMP)-2 and MMP9, and their tissue inhibitors TIMP1 and TIMP2 in patients with cervical cancer, patients with high-grade cervical intraepithelial dysplasia (CIN3) and patients with ectropion. PATIENTS AND METHODS: Concentration and expression of all tested parameters was measured in serum with enzyme-linked immunosorbent assay (ELISA) and in tissue with immunohistochemistry method. RESULTS: The epithelial expression of M-CSF and TIMP1 in cancer tissue was much stronger as compared to that in ectropion and CIN3. In the case of MMP2, lack of or weak expression in epithelial cells was observed in all tested groups. Our studies showed statistical differences of tested parameters in tissue expression and in plasma concentrations in patients with cervical cancer, patients with CIN3 and patients with ectropion. Moreover, data revealed positive correlation between plasma level and cervical cancer cell expression of VEGF. CONCLUSION: Our findings indicate a potential role of all the proteins tested here in cervical cancer diagnosis, especially VEGF. However, further studies will show whether they play a role in the progression of cancerous changes in epithelial tissue of the cervix.


Assuntos
Fator Estimulador de Colônias de Macrófagos/análise , Metaloproteases/análise , Inibidores Teciduais de Metaloproteinases/análise , Displasia do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/química , Fator A de Crescimento do Endotélio Vascular/análise , Adenocarcinoma/sangue , Adenocarcinoma/química , Adulto , Carcinoma de Células Escamosas/sangue , Carcinoma de Células Escamosas/química , Citocinas/análise , Citocinas/sangue , Feminino , Humanos , Fator Estimulador de Colônias de Macrófagos/sangue , Metaloproteinase 2 da Matriz/análise , Metaloproteinase 2 da Matriz/sangue , Metaloproteinase 9 da Matriz/análise , Metaloproteinase 9 da Matriz/sangue , Metaloproteases/sangue , Pessoa de Meia-Idade , Inibidor Tecidual de Metaloproteinase-1/análise , Inibidor Tecidual de Metaloproteinase-1/sangue , Inibidor Tecidual de Metaloproteinase-2/análise , Inibidor Tecidual de Metaloproteinase-2/sangue , Inibidores Teciduais de Metaloproteinases/sangue , Displasia do Colo do Útero/sangue , Neoplasias do Colo do Útero/sangue , Fator A de Crescimento do Endotélio Vascular/sangue , Adulto Jovem
6.
Chem Commun (Camb) ; 55(91): 13729-13732, 2019 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-31661085

RESUMO

A "signal-on" photoelectrochemical (PEC) biosensor was developed for ultrasensitive detection of vascular endothelial growth factor 165 (VEGF165) based on a Ti3C2/BiVO4 Schottky junction, which was a novel sensitized structure with excellent photovoltaic performance, prominent film-forming ability and stability. The generation of a built-in electric field by the Ti3C2/BiVO4 Schottky junction efficiently suppressed the combination of photogenerated electron-hole pairs and enhanced the photocurrent, providing a high initial photocurrent signal for the construction of a photoelectrochemical (PEC) biosensor.


Assuntos
Técnicas Biossensoriais/métodos , Bismuto/química , Titânio/química , Vanadatos/química , Fator A de Crescimento do Endotélio Vascular/análise , Aptâmeros de Nucleotídeos/química , Aptâmeros de Nucleotídeos/metabolismo , Técnicas Eletroquímicas , Limite de Detecção , Fator A de Crescimento do Endotélio Vascular/genética
7.
Braz Oral Res ; 33: e059, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31664357

RESUMO

We recently demonstrated that a co-culture system of human umbilical vein endothelial cells (HUVECs) and human dental pulp stem cells (hDPSCs) could enhance angiogenesis ability in vitro. However, whether tumor necrosis factor α (TNF-α) could promote blood vessel formation during pulp regeneration remained unknown. The aim of this study was to investigate the effects of TNF-α on the formation of endothelial tubules and vascular networks in a co-culture system of hDPSCs and HUVECs. hDPSCs were co-cultured with HUVECs at a ratio of 1:5. The Matrigel assay was performed to detect the total tubule branching lengths and numbers of branches, and the Cell-Counting Kit 8 assay was performed to examine the effect of TNF-α on cell proliferation. Real-time polymerase chain reactions and western blot were used to detect vascular endothelial growth factor (VEGF) mRNA and protein expression. The Matrigel assay showed significantly greater total branching lengths and numbers of branches formed in the experimental groups treated with different concentrations of TNF-α compared with the control group. The decomposition times of the tubule structures were also significantly prolonged (P < 0.05). Treatment with 50 ng/ml TNF-α did not significantly change the proliferation of co-cultured cells, but it significantly increased the VEGF mRNA and protein expression levels (p < 0.05). In addition, the migration abilities of HUVECs and hDPSCs increased after co-culture with TNF-α (p < 0.05). TNF-α enhanced angiogenic ability in vitro in the co-culture system of hDPSCs and HUVECs.


Assuntos
Indutores da Angiogênese/farmacologia , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologia , Adolescente , Adulto , Western Blotting , Contagem de Células , Ensaios de Migração Celular , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/fisiologia , Células Cultivadas , Colágeno , Polpa Dentária/fisiologia , Combinação de Medicamentos , Células Endoteliais da Veia Umbilical Humana/fisiologia , Humanos , Laminina , Neovascularização Fisiológica/fisiologia , Proteoglicanas , Reação em Cadeia da Polimerase em Tempo Real , Valores de Referência , Reprodutibilidade dos Testes , Fatores de Tempo , Fator A de Crescimento do Endotélio Vascular/análise , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacos , Adulto Jovem
8.
J Appl Oral Sci ; 27: e20180671, 2019 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-31508795

RESUMO

OBJECTIVE: To monitor early periodontal disease progression and to investigate clinical and molecular profile of inflamed sites by means of crevicular fluid and gingival biopsy analysis. METHODOLOGY: Eighty-one samples of twenty-seven periodontitis subjects and periodontally healthy individuals were collected for the study. Measurements of clinical parameters were recorded at day -15, baseline and 2 months after basic periodontal treatment aiming at monitoring early variations ofthe clinical attachment level. Saliva, crevicular fluid and gingival biopsies were harvested from clinically inflamed and non-inflamed sites from periodontal patients and from control sites of healthy patients for the assessment of IL-10, MMP-8, VEGF, RANKL, OPG and TGF-ß1 protein and gene expression levels. RESULTS: Baseline IL-10 protein levels from inflamed sites were higher in comparison to both non-inflamed and control sites (p<0.05). Higher expression of mRNA for IL-10, RANK-L, OPG, e TGF-ß1 were also observed in inflamed sites at day -15 prior treatment (p<0.05). After the periodontal treatment and the resolution of inflammation, seventeen percent of evaluated sites still showed clinically detectable attachment loss without significant differences in the molecular profile. CONCLUSIONS: Clinical attachment loss is a negative event that may occur even after successful basic periodontal therapy, but it is small and limited to a small percentage of sites. Elevated inflammation markers of inflamed sites from disease patients reduced to the mean levels of those observed in healthy subjects after successful basic periodontal therapy. Significantly elevated both gene and protein levels of IL-10 in inflamed sites prior treatment confirms its modulatory role in the disease status.


Assuntos
Perda da Inserção Periodontal/patologia , Periodontite/patologia , Adulto , Biomarcadores/análise , Biópsia , Estudos de Casos e Controles , Citocinas/análise , Feminino , Gengiva/patologia , Líquido do Sulco Gengival/química , Humanos , Masculino , Metaloproteinase 8 da Matriz/análise , Pessoa de Meia-Idade , Osteoprotegerina/análise , Periodontite/terapia , Reação em Cadeia da Polimerase em Tempo Real , Saliva/química , Estatísticas não Paramétricas , Fatores de Tempo , Fator A de Crescimento do Endotélio Vascular/análise
9.
Biochim Biophys Acta Mol Cell Biol Lipids ; 1864(12): 158526, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31520775

RESUMO

Although human adipose tissue-derived stromal vascular fraction (SVF) has been considered a promising source of stem cells, its characteristics relevant to treatment of a damaged liver have not been fully elucidated. In the present study, we sought to characterize the property of human SVF and determine the therapeutic utility of SVF in the liver cirrhosis model. We performed microarray, quantitative (q)-PCR experiments, and in vivo therapeutic assays using a liver cirrhotic mouse model. q-PCR results revealed that hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF)-A, Interleukin (IL)-10 and microRNA (miR)-146 were more highly upregulated in SVF than in human adipose-derived mesenchymal stem cells (ASCs). The SVF culture medium (CM) inhibited the activation of hepatic stellate cells in vitro. Injection of SVF significantly suppressed TAA-induced liver fibrosis and repaired liver function by inhibition of infiltrating inflammatory cells and induction of capillary/hepatocyte regeneration in vivo. Injection of IL-10 siRNA treated SVF cells decreased anti-inflammation and anti-fibrotic effects in TAA-induced mice liver. Our data indicate that SVF show a high anti-inflammatory property for treating fibrotic liver diseases through IL-10 secretion. Therefore, SVF might be a novel therapeutic alternative for the treatment of liver cirrhosis in clinical settings.


Assuntos
Inflamação/terapia , Cirrose Hepática/terapia , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/imunologia , Animais , Linhagem Celular , Células Cultivadas , Modelos Animais de Doenças , Fator de Crescimento de Hepatócito/análise , Fator de Crescimento de Hepatócito/imunologia , Humanos , Inflamação/imunologia , Inflamação/fisiopatologia , Interleucina-10/análise , Interleucina-10/imunologia , Fígado/imunologia , Fígado/fisiopatologia , Cirrose Hepática/imunologia , Cirrose Hepática/fisiopatologia , Masculino , Células-Tronco Mesenquimais/citologia , Camundongos Endogâmicos NOD , Camundongos SCID , Fator A de Crescimento do Endotélio Vascular/análise , Fator A de Crescimento do Endotélio Vascular/imunologia
10.
Bull Hosp Jt Dis (2013) ; 77(3): 189-193, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31487484

RESUMO

BACKGROUND: Synovial fluid biomarkers can highlight the molecular milieu associated with knee pathology and have been shown to be significantly different in patients with anterior cruciate ligament (ACL) injuries compared to uninjured controls. The purpose of the current study was to establish how synovial fluid biomarker concentrations change in patients undergoing ACL reconstruction between the immediate preoperative period to the acute postoperative period. METHODS: Patients were prospectively enrolled at the time of surgery from September 2016 to March 2017. Patients who had an operative knee synovial fluid sample obtained at the time of ACL reconstruction and provided a synovial fluid sample at their first postoperative appointment were included. The concentrations of 10 biomarkers were determined using a multiplex magnetic bead immunoassay. Biomarker concentrations before and after surgery were compared using a paired sample t-test. RESULTS: Eight patients with mean age of 33.4 years who underwent isolated ACL reconstruction using a bonepatellar tendon-bone autograft were included. The mean time between surgery and postoperative office visit was 10.4 days. There was a statistically significant increase in the concentrations of interleukin-6 (IL-6, p = 0.014), monocyte chemoattractant protein-1 (MCP-1, p = 0.024), human matrix metalloproteinase 3 (MMP-3, p = 0.00002), macrophage inflammatory protein-1 beta (MIP-1ß, p = 0.006), human interleukin-1 receptor antagonist (IL-1Ra, p = 0.017), and vascular endothelial growth factor (VEGF, p = 0.023) between the time of surgery and the first postoperative visit and a decrease in the concentration of tissue inhibitor of metalloproteinase-2 (p = 0.050). CONCLUSION: The molecular profile of the synovial fluid changes in the early postoperative period following arthroscopic ACL reconstruction. The concentration of proinflammatory markers (such as IL-6, MCP-1, MMP-3, and MIP-1ß) and growth factors including VEGF increases. The concentration of the anti-inflammatory marker tissue inhibitor of metalloproteinase-2 (TIMP-2) appears to decrease postoperatively.


Assuntos
Lesões do Ligamento Cruzado Anterior/cirurgia , Biomarcadores/análise , Líquido Sinovial , Adulto , Reconstrução do Ligamento Cruzado Anterior/efeitos adversos , Reconstrução do Ligamento Cruzado Anterior/métodos , Artroscopia/métodos , Quimiocina CCL2/análise , Quimiocina CCL4/análise , Feminino , Humanos , Proteína Antagonista do Receptor de Interleucina 1/análise , Interleucina-6/análise , Articulação do Joelho/imunologia , Articulação do Joelho/metabolismo , Articulação do Joelho/cirurgia , Masculino , Metaloproteinase 3 da Matriz/análise , Período Perioperatório , Líquido Sinovial/imunologia , Líquido Sinovial/metabolismo , Fator A de Crescimento do Endotélio Vascular/análise
11.
Nucleic Acids Res ; 47(16): 8362-8374, 2019 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-31392985

RESUMO

Many nucleic acid aptamers that bind to target molecules have been reported as antibody alternatives. However, while the affinities of aptamers vary widely, little is known about the relationship between the affinities and their applicabilities for practical use. Here, we developed molecular affinity rulers: a series of DNA aptamers with different affinities that bind to the same area of target molecules, to measure the aptamer and its device applicabilities. For the ruler preparation, we used high-affinity DNA aptamers containing a hydrophobic unnatural base (Ds) as the fifth base. By replacing Ds bases with A bases in Ds-DNA aptamers targeting VEGF165 and interferon-γ, we prepared two sets of DNA aptamers with dissociation constants (KD) ranging from 10-12 to 10-8 M. Using these molecular affinity rulers, we evaluated the sensitivity of DNA aptamers in ELISA (enzyme-linked immunosorbent assay), which showed the clear relationship between aptamer affinities and their detection sensitivities. In sandwich-type ELISA using combinations of aptamers and antibodies, aptamers with KD values lower than ∼10-9 M were required for sufficient sensitivities (limit of detection (LOD) < 10 pM) and signal intensities, but optimizations improved the lower-affinity aptamers' applicabilities. These aptamer affinity rulers could be useful for evaluating and improving aptamer applicabilities.


Assuntos
Adenina/química , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais , Ensaio de Imunoadsorção Enzimática/métodos , Interferon gama/análise , Fator A de Crescimento do Endotélio Vascular/análise , Animais , Anticorpos Monoclonais/química , Aptâmeros de Nucleotídeos/síntese química , Pareamento de Bases , Sequência de Bases , Biotina/química , Ensaio de Imunoadsorção Enzimática/normas , Humanos , Interações Hidrofóbicas e Hidrofílicas , Interferon gama/química , Cinética , Limite de Detecção , Conformação de Ácido Nucleico , Ligação Proteica , Padrões de Referência , Técnica de Seleção de Aptâmeros , Estreptavidina/química , Fator A de Crescimento do Endotélio Vascular/química
12.
Arterioscler Thromb Vasc Biol ; 39(8): 1652-1666, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31294623

RESUMO

OBJECTIVE: We examined the pathogenic significance of VEGF (vascular endothelial growth factor)-A in experimental abdominal aortic aneurysms (AAAs) and the translational value of pharmacological VEGF-A or its receptor inhibition in aneurysm suppression. Approaches and Results: AAAs were created in male C57BL/6J mice via intra-aortic elastase infusion. Soluble VEGFR (VEGF receptor)-2 extracellular ligand-binding domain (delivered in Ad [adenovirus]-VEGFR-2), anti-VEGF-A mAb (monoclonal antibody), and sunitinib were used to sequester VEGF-A, neutralize VEGF-A, and inhibit receptor tyrosine kinase activity, respectively. Influences on AAAs were assessed using ultrasonography and histopathology. In vitro transwell migration and quantitative reverse transcription polymerase chain reaction assays were used to assess myeloid cell chemotaxis and mRNA expression, respectively. Abundant VEGF-A mRNA and VEGF-A-positive cells were present in aneurysmal aortae. Sequestration of VEGF-A by Ad-VEGFR-2 prevented AAA formation, with attenuation of medial elastolysis and smooth muscle depletion, mural angiogenesis and monocyte/macrophage infiltration. Treatment with anti-VEGF-A mAb prevented AAA formation without affecting further progression of established AAAs. Sunitinib therapy substantially mitigated both AAA formation and further progression of established AAAs, attenuated aneurysmal aortic MMP2 (matrix metalloproteinase) and MMP9 protein expression, inhibited inflammatory monocyte and neutrophil chemotaxis to VEGF-A, and reduced MMP2, MMP9, and VEGF-A mRNA expression in macrophages and smooth muscle cells in vitro. Additionally, sunitinib treatment reduced circulating monocytes in aneurysmal mice. CONCLUSIONS: VEGF-A and its receptors contribute to experimental AAA formation by suppressing mural angiogenesis, MMP and VEGF-A production, myeloid cell chemotaxis, and circulating monocytes. Pharmacological inhibition of receptor tyrosine kinases by sunitinib or related compounds may provide novel opportunities for clinical aneurysm suppression.


Assuntos
Aneurisma da Aorta Abdominal/etiologia , Elastase Pancreática/farmacologia , Receptores de Fatores de Crescimento do Endotélio Vascular/fisiologia , Fator A de Crescimento do Endotélio Vascular/fisiologia , Animais , Aneurisma da Aorta Abdominal/tratamento farmacológico , Aneurisma da Aorta Abdominal/metabolismo , Quimiotaxia/efeitos dos fármacos , Modelos Animais de Doenças , Progressão da Doença , Metaloproteinase 2 da Matriz/análise , Metaloproteinase 9 da Matriz/análise , Camundongos , Receptores de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Sunitinibe/uso terapêutico , Fator A de Crescimento do Endotélio Vascular/análise , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores
13.
Chem Commun (Camb) ; 55(56): 8076-8078, 2019 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-31225560

RESUMO

We developed a novel "signal-on" photoelectrochemical (PEC) aptasensor with a near-zero background signal by using AgVO3 as a single photoactive material for the sensitive detection of vascular endothelial growth factor (VEGF165).


Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas/métodos , Óxidos/química , Processos Fotoquímicos , Prata/química , Compostos de Vanádio/química , Fator A de Crescimento do Endotélio Vascular/análise , Humanos , Limite de Detecção
14.
Folia Med (Plovdiv) ; 61(1): 61-68, 2019 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31237859

RESUMO

BACKGROUND: Thyroid carcinomas are the most common endocrine tumors - they account for 95% of all endocrine tumors. Thyroid carcinomas are more vascular than normal thyroid tissue. For the tumor to grow and subsequently metastasize it is crucial that it induces angiogenesis. This requires a change in the balance between certain angiogenic factors such as the vascular endothelial growth factor-A (VEGF-A) and the inhibitors of angiogenesis. The aim of this study was to evaluate the role of VEGF-A expression in thyroid carcinomas. MATERIALS AND METHODS: The present prospective study included 80 cases, of which 60 were patients with thyroid cancer including papillary, follicular, medullary and anaplastic carcinoma, and 20 patients (controls) with benign thyroid tissue (thyroid goiter). All cases were examined using the immunohistochemical staining for VEGF-A. RESULTS: VEGF-A expression in thyroid carcinoma was significantly higher than in benign thyroid tissues (p<0.001). VEGF-A expression values in thyroid carcinoma did not associate with tumor necrosis degree (p=1.000). Furthermore, VEGF-A expression values in thyroid carcinoma were not associated with other prognostic factors such as tumor hemorrhage, angioinvasion, atypical mitosis, and lymphatic invasion. CONCLUSION: Our data showed that the VEGF-A expression is upregulated in thyroid cancers compared with benign thyroid tissue. Therefore, it would be useful to perform IHC staining for VEGF-A expression as a valuable diagnostic tool in TC.


Assuntos
Neoplasias da Glândula Tireoide/patologia , Fator A de Crescimento do Endotélio Vascular/fisiologia , Adulto , Idoso , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Necrose , Prognóstico , Estudos Prospectivos , Neoplasias da Glândula Tireoide/química , Fator A de Crescimento do Endotélio Vascular/análise
15.
Anticancer Res ; 39(6): 3147-3157, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31177161

RESUMO

BACKGROUND/AIM: Regional lymph node recurrence (RLNR) is the most common pattern of recurrence within 2 years from the diagnosis of patients with non-metastatic malignant cutaneous melanoma. However, isolated RLNR without distant metastasis has been rarely studied. PATIENTS AND METHODS: Forty patients with isolated RLNR as a first recurrence were analyzed retrospectively. The clinical outcomes and prognostic impact of clinicopathologic parameters were analyzed. Immunostaining for FOXP3, VEGF, pAKT, and pS6 was also performed. RESULTS: The median disease-free interval from first diagnosis to isolated RLNR and post-recurrence recurrence-free survival (pRFS) were 12 months and 7.2 months, respectively. Distant failure was the most common pattern of failure after isolated RLNR (67.5%). The number of initially harvested lymph nodes (LN) >7 and LN ratio >22.2% at the time of recurrence were prognosticators for pRFS in multivariate analysis. None of the tested biomarkers were significantly related to prognosis. The 5-year post-recurrence overall survival rate was 84.9%. CONCLUSION: Most patients with isolated RLNR will experience a second failure within months, especially distantly. The number of initially harvested LNs and LN ratio at the time of recurrence could predict pRFS.


Assuntos
Melanoma/secundário , Neoplasias Cutâneas/patologia , Adolescente , Adulto , Idoso , Biomarcadores Tumorais/análise , Bases de Dados Factuais , Progressão da Doença , Feminino , Fatores de Transcrição Forkhead/análise , Humanos , Excisão de Linfonodo , Metástase Linfática , Masculino , Melanoma/química , Melanoma/mortalidade , Melanoma/cirurgia , Pessoa de Meia-Idade , Fosforilação , Intervalo Livre de Progressão , Proteínas Proto-Oncogênicas c-akt/análise , Estudos Retrospectivos , Proteína S6 Ribossômica/análise , Fatores de Risco , Neoplasias Cutâneas/química , Neoplasias Cutâneas/mortalidade , Neoplasias Cutâneas/cirurgia , Fatores de Tempo , Fator A de Crescimento do Endotélio Vascular/análise , Adulto Jovem
16.
BMC Ophthalmol ; 19(1): 128, 2019 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-31185956

RESUMO

BACKGROUND: The aim of the present study was to evaluate the efficacy and safety of intravitreal conbercept combined with trabeculectomy and panretinal photocoagulation for neovascular glaucoma (NVG). METHODS: Fifty patients (54 eyes) with NVG were included in this prospective study. Fifty-two eyes initially underwent intravitreal conbercept (0.5 mg/0.05 ml) treatment followed by trabeculectomy and panretinal photocoagulation. Preoperative and postoperative best-corrected visual acuity (BCVA), intraocular pressure (IOP), the number of antiglaucoma medications, and surgical complications were recorded. The levels of VEGF-A, TGF-ß1 and PLGF in aqueous humour samples collected during surgery were measured by enzyme-linked immunosorbent assay (ELISA). Light microscopy and transmission electron microscopy were used to observe the surgically excised trabecular tissue; enucleation was performed in 2 eyes, and light microscopy was used as the histopathological control. RESULTS: The follow-up period after trabeculectomy was 1 year. Of the 52 eyes, 39 completed 1 year of follow-up, and 13 were lost to follow-up. Recurrence of iris neovascularization was observed in 5 eyes, 9 had hyphema, 16 had filter-bled scarring, and no eye had complications attributable to the drug. The mean IOP was reduced from 48.1 ± 14.2 to 23.2 ± 8.7 mmHg, and the mean number of antiglaucoma medications used decreased from 3.0 (3.0, 4.0) to 1.0 (0.0, 1.0) after 1 year (both P < 0.05). The complete success rate was 76.9, 76.9, 71.0, 51.6, and 32.3% at 1 week, 1 month, 3 months, 6 months and 12 months, respectively, when the cut-off IOP was 18 mmHg. After patients underwent intravitreal injection, the concentrations of VEGF-A and TGF-ß1 in the aqueous humour in NVG patients decreased from 168.8 ± 13.4 and 159.6 ± 15.4 pg/ml to 160.2 ± 7.6 and 151.9 ± 2.3 pg/ml, respectively (both P < 0.05). Light microscopy revealed neovascularization regression in the iris in specimens treated with intravitreal conbercept. Electron microscopy revealed trabecular endothelial cell degeneration in the conbercept-treated specimens. CONCLUSIONS: Our initial findings suggest that intravitreal conbercept is an effective treatment for managing NVG that has fewer short-term postoperative complications. TRIAL REGISTRATION: Current Controlled Trials ChiCTR1800019918 , 8 December 2018, retrospectively registered.


Assuntos
Glaucoma Neovascular/tratamento farmacológico , Proteínas Recombinantes de Fusão/administração & dosagem , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Humor Aquoso/metabolismo , Feminino , Glaucoma Neovascular/metabolismo , Glaucoma Neovascular/fisiopatologia , Humanos , Pressão Intraocular/fisiologia , Injeções Intravítreas , Fotocoagulação , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Trabeculectomia , Fator de Crescimento Transformador beta1/análise , Fator A de Crescimento do Endotélio Vascular/análise , Acuidade Visual/fisiologia , Adulto Jovem
17.
J Clin Pathol ; 72(9): 615-621, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31235542

RESUMO

AIMS: Serous cystadenomata (SCAs) are benign pancreatic cystic neoplasms that present a diagnostic challenge despite many investigational approaches. Notwithstanding the promise of molecular diagnostics, these tests have limited accessibility in day-to-day surgical pathology practices. We aim to corroborate and build on recent evidence which suggests that positive α-inhibin immunohistochemistry (IHC) is a helpful adjunct in the biopsy confirmation of pancreatic SCA. METHODS: We retrospectively reviewed 22 fine-needle aspirates/biopsies from 14 patients (mean age 65 years, 47-83 years) with pancreatic multicystic lesions radiologically suspicious for SCA (location: 6 body, 2 head, 4 tail, 1 neck, 1 uncinate; cyst size: mean 3.7 cm, 2.0-7.6 cm), as well as an additional 10 pancreatic resection specimens with confirmed SCA; α-inhibin IHC was performed on all cell blocks, biopsy slides and representative resection specimen sections. Where available, associated cyst fluid was analysed for correlative vascular endothelial growth factor A (VEGF-A) and carcinoembryonic antigen levels. RESULTS: An α-inhibin IHC sensitivity of 80% was observed in the cases with resection confirmed SCA. Of the fine-needle aspirate/biopsy specimens, 59% (13/22) contained epithelial cells strongly positive for α-inhibin. When selecting for specimens that exhibited distinct strips of epithelium, the α-inhibin strong positivity rate increased to 73% (8/11). VEGF-A values were supportive of false-negative α-inhibin IHC in three cases and true-negative α-inhibin IHC in one case. CONCLUSION: This study postulates a diagnostic algorithm to confirm pancreatic SCA which may help to decrease unnecessary follow-up endoscopy/surgical resection and would decrease the associated morbidity, mortality and financial costs in patients with this otherwise benign condition.


Assuntos
Biomarcadores Tumorais/análise , Cistadenoma Seroso/química , Cistadenoma Seroso/patologia , Imuno-Histoquímica , Inibinas/análise , Neoplasias Pancreáticas/química , Neoplasias Pancreáticas/patologia , Idoso , Idoso de 80 Anos ou mais , Algoritmos , Biópsia por Agulha Fina , Antígeno Carcinoembrionário/análise , Cistadenoma Seroso/cirurgia , Técnicas de Apoio para a Decisão , Árvores de Decisões , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Pancreáticas/cirurgia , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Estudos Retrospectivos , Fator A de Crescimento do Endotélio Vascular/análise
18.
Dalton Trans ; 48(26): 9824-9830, 2019 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-31147654

RESUMO

Vascular endothelial growth factor (VEGF165), an important glycosylated protein from the VEGF family, is a type of signal protein highly associated with the development and progression of cancers. In this work, we designed a G-quadruplex-based aptasensing platform for the sensitive and selective detection of VEGF165 in aqueous solution and red blood cell solution. A long-lived phosphorescence iridium(iii) complex (1) with promising photophysical properties and a large Stokes shift was chosen as a selective G-quadruplex probe. The platform could achieve a limit of detection (LOD) down to the picomolar level using a conventional fluorometer. Furthermore, we successfully applied the platform to a three-step suspended droplet (SD)-based microfluidic device for the monitoring of VEGF165. In contrast to the channel-based and digital microfluidic chips, SD-based chips allow easy introduction of liquid samples, valve-free manipulation of multiple reaction steps and flexible volume range. Importantly, polypropylene (PP), a hydrophobic and thermally stable material, was chosen as a substrate to fabricate the chip for the SD-based microfluidic device. The PP-based chip allows the combination of superhydrophobic force, gravity and surface tension for effective driving of the suspended droplet throughout the channel without reverse migration. After assembling all the major components, including a UV lamp, a rotatable chip holder, a filter and a camera into the portable device, we successfully demonstrated the applicability of the device to detect VEGF165 in aqueous solution with a LOD of 0.33 nM at a signal-to-noise ratio (S/N) of 3 and a linear range of 1-100 nM.


Assuntos
Técnicas Analíticas Microfluídicas , Oligonucleotídeos/química , Fator A de Crescimento do Endotélio Vascular/análise , Humanos , Estrutura Molecular , Tamanho da Partícula
19.
Korean J Parasitol ; 57(2): 117-125, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31104403

RESUMO

Malarial infection induces tissue hypoxia in the host through destruction of red blood cells. Tissue hypoxia in malarial infection may increase the activity of HIF1α through an intracellular oxygen-sensing pathway. Activation of HIF1α may also induce vascular endothelial growth factor (VEGF) to trigger angiogenesis. To investigate whether malarial infection actually generates hypoxia-induced angiogenesis, we analyzed severity of hypoxia, the expression of hypoxia-related angiogenic factors, and numbers of blood vessels in various tissues infected with Plasmodium berghei. Infection in mice was performed by intraperitoneal injection of 2×106 parasitized red blood cells. After infection, we studied parasitemia and survival. We analyzed hypoxia, numbers of blood vessels, and expression of hypoxia-related angiogenic factors including VEGF and HIF1α. We used Western blot, immunofluorescence, and immunohistochemistry to analyze various tissues from Plasmodium berghei-infected mice. In malaria-infected mice, parasitemia was increased over the duration of infection and directly associated with mortality rate. Expression of VEGF and HIF1α increased with the parasitemia in various tissues. Additionally, numbers of blood vessels significantly increased in each tissue type of the malaria-infected group compared to the uninfected control group. These results suggest that malarial infection in mice activates hypoxia-induced angiogenesis by stimulation of HIF1α and VEGF in various tissues.


Assuntos
Células Endoteliais/patologia , Hipóxia , Malária/patologia , Neovascularização Patológica , Plasmodium berghei/crescimento & desenvolvimento , Animais , Western Blotting , Modelos Animais de Doenças , Feminino , Subunidade alfa do Fator 1 Induzível por Hipóxia/análise , Imuno-Histoquímica , Camundongos Endogâmicos C57BL , Microscopia de Fluorescência , Parasitemia/parasitologia , Análise de Sobrevida , Fator A de Crescimento do Endotélio Vascular/análise
20.
Life Sci ; 230: 111-120, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31129141

RESUMO

AIMS: In women, uterine alterations have been associated with sex steroid hormones. Sex hormones regulate the expression of thyroid hormone receptors (TRs) in the uterus, but an inverse link is unknown. We analyzed the impact of hypothyroidism on histological characteristics, vascular endothelial growth factor (VEGF-A), progesterone receptors (PR), estrogen receptors (ER), thyroid hormone receptors (TRs), perilipin (PLIN-A), and lipid content in the uterus of virgin rabbits. MAIN METHODS: Twelve Chinchilla-breed adult female rabbits were grouped into control (n = 6) and hypothyroid (n = 6; 0.02% of methimazole for 30 days). The thickness of endometrium and myometrium, number of uterine glands, and infiltration of immune cells were analyzed. The expression of VEGF-A, PR, ERα, and PLIN-A was determined by RT-PCR and western blot. The uterine content of triglycerides (TAG), total cholesterol (TC), and malondialdehyde (MDA) was quantified. KEY FINDINGS: Hypothyroidism promoted uterine hyperplasia and a high infiltration of immune cells into the endometrium, including macrophages CD163+. It also increased the expression of VEGF-A, TRA, and ERα-66 but reduced that of PR and ERα-46. The uterine content of PLIN-A, TAG, and TC was reduced, but that of MDA was augmented in hypothyroid rabbits. SIGNIFICANCE: Our results suggest that uterine hyperplasia and inflammation promoted by hypothyroidism should be related to changes in the VEGF-A, PR, ER, and TRs expression, as well as to modifications in the PLIN-A expression, lipid content, and oxidative status. These results suggest that hypothyroidism should affect the fertility of females.


Assuntos
Hormônios Esteroides Gonadais/metabolismo , Hiperplasia/etiologia , Hiperplasia/fisiopatologia , Hipotireoidismo/complicações , Animais , Endométrio/metabolismo , Feminino , Expressão Gênica/efeitos dos fármacos , Hormônios Esteroides Gonadais/análise , Hipotireoidismo/fisiopatologia , Inflamação , Lipídeos/análise , Miométrio/metabolismo , Perilipina-1/análise , Perilipina-1/metabolismo , Progesterona/farmacologia , Coelhos , Receptores Estrogênicos/análise , Receptores Estrogênicos/metabolismo , Receptores de Progesterona/análise , Receptores de Progesterona/metabolismo , Receptores dos Hormônios Tireóideos/análise , Receptores dos Hormônios Tireóideos/metabolismo , Útero/metabolismo , Útero/fisiologia , Fator A de Crescimento do Endotélio Vascular/análise
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