RESUMO
Health interventions often depend on a complex system of human and capital infrastructure that is shared with other interventions, in the form of service delivery platforms, such as healthcare facilities, hospitals, or community services. Most forms of health system strengthening seek to improve the efficiency or effectiveness of such delivery platforms. This paper presents a typology of ways in which health system strengthening can improve the economic efficiency of health services. Three types of health system strengthening are identified and modelled: (1) investment in the efficiency of an existing shared platform that generates positive benefits across a range of existing interventions; (2) relaxing a capacity constraint of an existing shared platform that inhibits the optimization of existing interventions; (3) providing an entirely new shared platform that supports a number of existing or new interventions. Theoretical models are illustrated with examples, and illustrate the importance of considering the portfolio of interventions using a platform, and not just piecemeal individual analysis of those interventions. They show how it is possible to extend principles of conventional cost-effectiveness analysis to identify an optimal balance between investing in health system strengthening and expenditure on specific interventions. The models developed in this paper provide a conceptual framework for evaluating the cost-effectiveness of investments in strengthening healthcare systems and, more broadly, shed light on the role that platforms play in promoting the cost-effectiveness of different interventions.
Assuntos
Análise Custo-Benefício , Atenção à Saúde , Programas Governamentais , Humanos , Modelos TeóricosRESUMO
Myofibroblasts represent specific subpopulations of cells with important roles in tissue remodeling in both health and disease. They are not usually found in resting healthy tissues. However, they increase in number during the proliferative phase of wound healing. In these conditions, myofibroblasts secrete and organize different molecular components of the extracellular matrix that with time will reconstitute and hopefully regenerate the damaged tissue. Importantly, these cell populations must be eliminated after wound healing has been completed. However, deficiencies in their differentiation or the persistence of this cell population has been associated with the development of delayed wound healing and fibrosis, respectively. In the present review, we analyze the involvement of myofibroblasts in periodontal wound healing and their potential contribution to tissue homeostasis and disease.
Assuntos
Crescimento Excessivo da Gengiva/fisiopatologia , Miofibroblastos/fisiologia , Cicatrização , Homeostase , Humanos , Miofibroblastos/citologiaRESUMO
BACKGROUND AND OBJECTIVES: An important goal of periodontal therapy is the modulation of the inflammatory response. To this end, several pharmacological agents have been evaluated. Triclosan corresponds to an antibacterial and anti-inflammatory agent currently used in periodontal therapy. Chitosan is a natural polymer that may act as a drug delivery agent and exerts antibacterial and anti-inflammatory activities. Therefore, an association between both molecules might be useful to prevent inflammation and tissue destruction in periodontal tissues. MATERIAL AND METHODS: In the present study, we have generated chitosan-triclosan particles and evaluated their morphology, charge, biocompatibility and gene expression analysis in human gingival fibroblasts. RESULTS: The chitosan-triclosan particles size and Z potential were 129 ± 47 nm and 51 ± 17 mV respectively. Human gingival fibroblast viability was not affected by chitosan-triclosan. A total of 1533 genes were upregulated by interleukin (IL)-1ß. On the other hand, 943 were downregulated in fibroblasts stimulated with IL-1ß plus chitosan-triclosan particles. Fifty-one genes were identified as molecular targets upregulated by IL-1 ß and downregulated by the chitosan-triclosan particles. The gene ontology analysis revealed that these genes were enriched in categories related to biological processes, molecular function and cellular components. Furthermore, using real-time reverse transcription-polymerase chain reaction beta-actin, fibronectin, interleukin-6 and IL-1b genes were confirmed as targets upregulated by IL-1ß and downregulated by chitosan-triclosan particles. CONCLUSION: Our results show that chitosan-triclosan particles are able to modulate the inflammatory response in gingival fibroblasts. This effect might be useful in the prevention and/or treatment of inflammation in periodontal diseases.
Assuntos
Anti-Inflamatórios/farmacologia , Quitosana/farmacologia , Fibroblastos/efeitos dos fármacos , Gengiva/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Triclosan/farmacologia , Adolescente , Adulto , Antibacterianos/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Chile , Quitosana/química , Ciclo-Oxigenase 2/genética , Regulação para Baixo/efeitos dos fármacos , Expressão Gênica , Ontologia Genética , Humanos , Inflamação/tratamento farmacológico , Inflamação/prevenção & controle , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Masculino , Teste de Materiais , Dente Molar , Tamanho da Partícula , Doenças Periodontais/tratamento farmacológico , Doenças Periodontais/prevenção & controle , Periodonto/efeitos dos fármacos , RNA/análise , RNA/isolamento & purificação , Triclosan/química , Regulação para Cima/efeitos dos fármacos , Adulto JovemRESUMO
The WHO Commission on the Social Determinants of Health set out an impressive collection of policy proposals on the social determinants of health. However, a serious weakness for securing implementation is the difficulty for policymakers in identifying priorities for action. The objective of this study is to determine a small set of the most influential determinants using existing data and an empirical approach. 45 Indicators from the World Bank's World Development Indicators are selected to measure attainment for the determinants proposed by the Commission. Panel data models of life expectancy at birth for 54 low-income countries over the years 1990-2012 (1188 country-years) are estimated. Each determinant is subjected to a robustness test using Extreme Bound Analysis, to determine the stability of its estimated impact on life expectancy. For 20 robust and significant determinants the magnitude of association with life expectancy is determined. The largest average increases in life expectancy at 14.5 months per capita is associated with a one standard deviation reduction in HIV prevalence among children, followed by advances in gender equality at 9.4 months. Improvements in life expectancy between 6 and 9 months are associated with agricultural production, political stability, access to clean water and sanitation, good governance, and primary school enrolment. Improvements below 6 months are associated with increases in private health expenditure and overseas development assistance, and control of armed conflict and HIV prevalence among men. There is no evidence that national income, public spending on healthcare and education, secondary schooling, terms of international trade, employment, debt service and relief, out-of-pocket expenditures, agricultural ex- or imports, lifestock production, foreign investment, urbanization or environmental degradation are robustly associated with population health. Results provide support for the relevance of some proposed policies. The findings can inform priorities for future research and policy action on the social determinants of health.
Assuntos
Países em Desenvolvimento/estatística & dados numéricos , Política de Saúde , Prioridades em Saúde/tendências , Expectativa de Vida/tendências , Determinantes Sociais da Saúde , Escolaridade , Gastos em Saúde/estatística & dados numéricos , Humanos , Pobreza/estatística & dados numéricos , Pobreza/tendências , Análise de Regressão , Fatores SocioeconômicosRESUMO
BACKGROUND AND OBJECTIVES: Methylglyoxal is a toxic product derived from glucose metabolism that plays a role in inflammation, diabetes and aging. In addition, the periodontal pathogen Tannerella forsythensis may also generate this compound. However, the effects of methylglyoxal on gingival cells are still poorly understood. In the present study, we have explored whether methylglyoxal or methylglyoxal-treated collagen may modulate cell viability, death and proliferation in gingival connective tissue cells. In addition, we have searched for inflammatory mediators secreted by cells upon exposure to these conditions. MATERIAL AND METHODS: Primary cultures of human gingival fibroblasts were stimulated with soluble methylglyoxal or cultured over a collagen matrix glycated by this agent. Cell viability was evaluated through the MTS assay. Cell death was assessed through DAPI nuclear staining, annexin V and propidium iodide assays. Cell proliferation was evaluated through double immunofluorescence for DAPI and Ki67. Protein levels of matrix metalloproteinases and cytokines were assessed through antibody arrays, enzyme-linked immunosorbent assay, real-time reverse transcription polymerase chain reaction and immunofluorescence. Statistical analysis was performed using the Kruskall-Wallis and Mann-Whitney tests. RESULTS: Soluble methylglyoxal, but not culture of gingival fibroblasts over a methylglyoxal-modified collagen matrix, induced a reduction on cell viability. Moreover, soluble methylglyoxal induced apoptotic cell death as indicated by DAPI nuclear staining, annexin V and propidium iodide assays. Neither soluble methylglyoxal, nor methylglyoxal-modified collagen modified cell proliferation. Using an antibody array, enzyme-linked immunosorbent assay and immunofluorescence assays, we determined that both, soluble methylglyoxal and methylglyoxal-modified collagen stimulated an increase in tissue inhibitor of metalloproteinase (TIMP)-1 protein levels. CONCLUSIONS: Soluble methylglyoxal is a highly cytotoxic compound that induces cell death through apoptosis in gingival fibroblasts. TIMP-1 is induced in these cells upon direct exposure to methylglyoxal or after culture of gingival fibroblasts over methylglyoxal-treated collagen. As TIMP-1 has been implicated in cell survival and matrix remodeling, we propose that increased TIMP-1 protein levels may be part of a protective response of gingival connective tissue cells upon exposure to methylglyoxal or after the interaction with the collagen matrix that has been modified by this agent.
Assuntos
Gengiva/efeitos dos fármacos , Aldeído Pirúvico/efeitos adversos , Adolescente , Adulto , Apoptose/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Colágeno/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Fibroblastos/efeitos dos fármacos , Imunofluorescência , Gengiva/citologia , Humanos , Masculino , Aldeído Pirúvico/metabolismo , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Adulto JovemRESUMO
There is a growing awareness amongst critical care practitioners that the impact of intensive care medicine extends beyond the patient to include the psychological impact on close family members. Several studies have addressed the needs of relatives within the intensive care context but the psychobiological impact of the experience has largely been ignored. Such impact is important in respect to health and well-being of the relative, with potential to influence patient recovery. The current feasibility study aimed to examine the acute psychobiological impact of the intensive care experience on relatives. Using a mixed methods approach, quantitative and qualitative data were collected simultaneously. Six relatives of patients admitted to the intensive care unit (ICU) of a District General Hospital, were assessed within 48 h of admission. Qualitative data were provided from semi-structured interviews analysed using interpretative phenomenological analysis. Quantitative data were collected using a range of standardised self-report questionnaires measuring coping responses, emotion, trauma symptoms and social support, and through sampling of diurnal salivary cortisol as a biomarker of stress. Four themes were identified from interview: the ICU environment, emotional responses, family relationships and support. Questionnaires identified high levels of anxiety, depression and trauma symptoms; the most commonly utilised coping techniques were acceptance, seeking support through advice and information, and substance use. Social support emerged as a key factor with focused inner circle support relating to family and ICU staff. Depressed mood and avoidance were linked to greater mean cortisol levels across the day. Greater social network and coping via self-distraction were related to lower evening cortisol, indicating them as protective factors in the ICU context. The experience of ICU has a psychological and physiological impact on relatives, suggesting the importance of identifying cost-effective interventions with evaluations of health benefits to both relatives and patients.
Assuntos
Ansiedade/epidemiologia , Cuidados Críticos/psicologia , Depressão/epidemiologia , Família/psicologia , Trauma Psicológico/epidemiologia , Estresse Psicológico/metabolismo , Estresse Psicológico/psicologia , Adaptação Psicológica , Adulto , Biomarcadores/metabolismo , Estudos de Viabilidade , Feminino , Humanos , Hidrocortisona/metabolismo , Unidades de Terapia Intensiva , Masculino , Pessoa de Meia-Idade , Pesquisa Qualitativa , Saliva/química , Apoio Social , Inquéritos e QuestionáriosRESUMO
Gingival wound healing comprises a series of sequential responses that allow the closure of breaches in the masticatory mucosa. This process is of critical importance to prevent the invasion of microbes or other agents into tissues, avoiding the establishment of a chronic infection. Wound healing may also play an important role during cell and tissue reaction to long-term injury, as it may occur during inflammatory responses and cancer. Recent experimental data have shown that gingival wound healing is severely affected by the aging process. These defects may alter distinct phases of the wound-healing process, including epithelial migration, granulation tissue formation, and tissue remodeling. The cellular and molecular defects that may explain these deficiencies include several biological responses such as an increased inflammatory response, altered integrin signaling, reduced growth factor activity, decreased cell proliferation, diminished angiogenesis, reduced collagen synthesis, augmented collagen remodeling, and deterioration of the proliferative and differentiation potential of stem cells. In this review, we explore the cellular and molecular basis of these defects and their possible clinical implications.
Assuntos
Envelhecimento/fisiologia , Gengiva/fisiologia , Cicatrização/fisiologia , Tecido Conjuntivo/fisiologia , Epitélio/fisiologia , Humanos , Inflamação/fisiopatologia , Peptídeos e Proteínas de Sinalização Intercelular/fisiologia , Neovascularização Fisiológica/fisiologiaRESUMO
BACKGROUND: Granulation tissue remodeling and myofibroblastic differentiation are critically important events during wound healing. Tobacco smoking has a detrimental effect in gingival tissue repair. However, studies evaluating the effects of cigarette smoke on these events are lacking. MATERIAL AND METHODS: We used gingival fibroblasts cultured within free-floating and restrained collagen gels to simulate the initial and final steps of the granulation tissue phase during tissue repair. Collagen gel contraction was stimulated with serum or transforming growth factor-ß1. Cigarette smoke condensate (CSC) was used to evaluate the effects of tobacco smoke on gel contraction. Protein levels of alpha-smooth muscle actin, ß1 integrin, matrix metalloproteinase-3 and connective tissue growth factor were evaluated through Western blot. Prostaglandin E(2) (PGE(2)) levels were determined through ELISA. Actin organization was evaluated through confocal microscopy. RESULTS: CSC reduced collagen gel contraction induced by serum and transforming growth factor-ß1 in restrained collagen gels. CSC also altered the development of actin stress fibers in fibroblasts cultured within restrained collagen gels. PGE(2) levels were strongly diminished by CSC in three-dimensional cell cultures. However, other proteins involved in granulation tissue remodeling and myofibroblastic differentiation such as alpha-smooth muscle actin, ß1 integrin, matrix metalloproteinase-3 and connective tissue growth factor, were unmodified by CSC. CONCLUSIONS: CSC may alter the capacity of gingival fibroblasts to remodel and contract a collagen matrix. Inhibition of PGE(2) production and alterations of actin stress fibers in these cells may impair proper tissue maturation during wound healing in smokers.
Assuntos
Dinoprostona/biossíntese , Fibroblastos/metabolismo , Gengiva/citologia , Nicotiana , Fumaça/efeitos adversos , Actinas/análise , Sangue , Sobrevivência Celular/fisiologia , Células Cultivadas , Colágeno Tipo I/efeitos dos fármacos , Colágeno Tipo I/metabolismo , Fator de Crescimento do Tecido Conjuntivo/análise , Citocalasina D/farmacologia , Dinoprostona/análise , Fibroblastos/efeitos dos fármacos , Géis , Gengiva/efeitos dos fármacos , Humanos , Integrina beta1/análise , Masculino , Metaloproteinase 3 da Matriz/análise , Nicotina/efeitos adversos , Técnicas de Cultura de Tecidos , Fator de Crescimento Transformador beta1/farmacologiaRESUMO
Aging may negatively affect gingival wound-healing. However, little is known about the mechanisms underlying this phenomenon. The present study examined the cellular responses associated with gingival wound-healing in aging. Primary cultures of human gingival fibroblasts were obtained from healthy young and aged donors for the analysis of cell proliferation, cell invasion, myofibroblastic differentiation, and collagen gel remodeling. Serum from young and old rats was used to stimulate cell migration. Gingival repair was evaluated in Sprague-Dawley rats of different ages. Data were analyzed by the Mann-Whitney and Kruskal-Wallis tests, with a p value of .05. Fibroblasts from aged donors showed a significant decrease in cell proliferation, migration, Rac activation, and collagen remodeling when compared with young fibroblasts. Serum from young rats induced higher cell migration when compared with serum from old rats. After TGF-beta1 stimulation, both young and old fibroblasts demonstrated increased levels of alpha-SMA. However, alpha-SMA was incorporated into actin stress fibers in young but not in old fibroblasts. After 7 days of repair, a significant delay in gingival wound-healing was observed in old rats. The present study suggests that cell migration, myofibroblastic differentiation, collagen gel remodeling, and proliferation are decreased in aged fibroblasts. In addition, altered cell migration in wound-healing may be attributable not only to cellular defects but also to changes in serum factors associated with the senescence process.
Assuntos
Envelhecimento/fisiologia , Gengiva/fisiologia , Actinas/efeitos dos fármacos , Envelhecimento/patologia , Animais , Diferenciação Celular/fisiologia , Movimento Celular/fisiologia , Proliferação de Células , Sobrevivência Celular/fisiologia , Células Cultivadas , Colágeno/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Fibroblastos/patologia , Fibroblastos/fisiologia , Gengiva/patologia , Gengivectomia/métodos , Humanos , Miofibroblastos/patologia , Miofibroblastos/fisiologia , Ratos , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta1/farmacologia , Cicatrização/fisiologia , Proteínas rac de Ligação ao GTP/análiseRESUMO
Chitosan is a naturally derived polymer with antimicrobial and anti-inflammatory properties. However, studies evaluating the role of chitosan in the control of periodontal pathogens and the responses of fibroblasts to inflammatory stimuli are lacking. In the present study, we analyzed whether chitosan particles may inhibit the growth of periodontal pathogens and modulate the inflammatory response in human gingival fibroblasts. Chitosan particles were generated through ionic gelation. They inhibited the growth of Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans at 5 mg/mL. Conversely, IL-1ß strongly stimulated PGE2 protein levels in gingival fibroblasts, and chitosan inhibited this response at 50 µg/mL. IL-1ß-stimulated PGE2 production was dependent on the JNK pathway, and chitosan strongly inhibited this response. IL-1ß stimulated NF-κB activation, another signaling pathway involved in PGE2 production. However, chitosan particles were unable to modify NF-κB signaling. The present study shows that chitosan exerts a predominantly anti-inflammatory activity by modulating PGE2 levels through the JNK pathway, which may be useful in the prevention or treatment of periodontal inflammation.
Assuntos
Aggregatibacter actinomycetemcomitans/efeitos dos fármacos , Antibacterianos/farmacologia , Anti-Inflamatórios/farmacologia , Quitosana/farmacologia , Gengiva/efeitos dos fármacos , Porphyromonas gingivalis/efeitos dos fármacos , Adulto , Antracenos/farmacologia , Técnicas Bacteriológicas , Sobrevivência Celular , Células Cultivadas , Dinoprostona/antagonistas & inibidores , Fibroblastos/efeitos dos fármacos , Gengiva/citologia , Humanos , Indicadores e Reagentes , Interleucina-1beta/farmacologia , Proteínas Quinases JNK Ativadas por Mitógeno/antagonistas & inibidores , L-Lactato Desidrogenase/análise , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , NF-kappa B/efeitos dos fármacos , Nanopartículas , Doenças Periodontais/microbiologia , Fosforilação , Transdução de Sinais/efeitos dos fármacos , Sais de Tetrazólio , Tiazóis , Fator de Transcrição RelA/efeitos dos fármacosRESUMO
The objective of the study was to review the literature reporting visual disturbance (VD)following sclerotherapy for varicose veins. Underlying mechanisms will be discussed. A literature search of the databases Medline and Google Scholar was performed. Original articles including randomized trials, case series and case reports reporting VD in humans following sclerotherapy for varicose veins were included. Additional references were also obtained if they had been referenced in related publications. The search yielded 4948 results of which 25 reports were found to meet the inclusion criteria. In larger series with at least 500 included patients the prevalence of VD following sclerotherapy ranges from 0.09% to 2%. In most reports foam sclerotherapy was associated with VD (19); exclusive use of liquid sclerosant was reported in two cases, some reports included foam and liquid sclerosant (4). There were no persistent visual disorders reported. VD occurred with polidocanol and sodium tetradecyl sulphate in different concentrations (0.253%). Various forms of foam preparation including various ways of foam production and the liquid air ratio (1 or 2 parts of liquid mixed with 3, 4 or 5 parts of air) were reported in association with the occurrence of VD. VDs following sclerotherapy for varicose veins are rare and all reported events were transient. Bubble embolism or any kind of embolism seems unlikely to be the only underlying mechanism. A systemic inflammatory response following sclerotherapy has been suggested. Further research to clarify the mechanism of action of sclerosants is required.
Assuntos
Polietilenoglicóis/efeitos adversos , Soluções Esclerosantes/efeitos adversos , Escleroterapia/efeitos adversos , Tetradecilsulfato de Sódio/efeitos adversos , Telangiectasia/terapia , Varizes/terapia , Transtornos da Visão/induzido quimicamente , Embolia/induzido quimicamente , Embolia/epidemiologia , Feminino , Humanos , MEDLINE , Masculino , Polidocanol , Polietilenoglicóis/uso terapêutico , Soluções Esclerosantes/uso terapêutico , Escleroterapia/métodos , Tetradecilsulfato de Sódio/uso terapêutico , Telangiectasia/epidemiologia , Varizes/epidemiologia , Transtornos da Visão/epidemiologiaRESUMO
BACKGROUND AND OBJECTIVE: Chitosan is a naturally derived polymer that may be applied in periodontal therapy for tissue-reconstruction purposes. Previous studies have shown that chitosan may stimulate tissue healing. However, reports exploring the cellular responses stimulated by chitosan are lacking. In the present study we analyzed whether chitosan may promote cell proliferation in primary cultures of human gingival fibroblasts. MATERIAL AND METHODS: Chitosan particles were generated, and their size, zeta potential and morphology were characterized using transmission and scanning electron microscopy and zetasizer analysis. The biocompatibility of chitosan particles was analyzed using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) cell-viability assay and by detecting the release of lactate dehydrogenase into the cell-culture medium. The total number of cells was estimated by staining with crystal violet followed by measurement of the absorbance at 560 nm on a microplate reader. Cell proliferation was studied by detecting proliferating cell nuclear antigen protein levels, immunofluorescence for Ki67 and incorporation of 5'-bromo-2'-deoxyuridine. RESULTS: The sizes of the chitosan particles generated were in the micrometer and nanometer ranges. Cell viability was increased in the presence of chitosan. Moreover, the combination of chitosan and platelet-derived growth factor (PDGF-BB) potently stimulated cell viability, cell proliferation and activation of the ERK1/2 pathway involved in cell proliferation. CONCLUSIONS: The present study shows that chitosan is well tolerated by gingival fibroblasts and is able to stimulate cell proliferation through the ERK1/2 signaling pathway. A synergistic response between chitosan and growth factors (such as PDGF-BB) may stimulate cell proliferation in gingival fibroblasts exposed to this biomaterial.
Assuntos
Indutores da Angiogênese/farmacologia , Materiais Biocompatíveis/farmacologia , Quitosana/farmacologia , Fibroblastos/efeitos dos fármacos , Gengiva/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-sis/farmacologia , Becaplermina , Materiais Biocompatíveis/química , Bromodesoxiuridina , Contagem de Células , Técnicas de Cultura de Células , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Quitosana/química , Corantes , Sinergismo Farmacológico , Violeta Genciana , Gengiva/citologia , Humanos , Antígeno Ki-67/análise , Antígeno Ki-67/efeitos dos fármacos , L-Lactato Desidrogenase/análise , L-Lactato Desidrogenase/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Nanopartículas/química , Tamanho da Partícula , Antígeno Nuclear de Célula em Proliferação/análise , Antígeno Nuclear de Célula em Proliferação/efeitos dos fármacos , Sais de Tetrazólio , TiazóisRESUMO
BACKGROUND AND OBJECTIVE: Several studies have analysed the role of nicotine as a prominent agent affecting wound repair in smokers. However, tobacco smoke contains several components that may alter gingival wound healing. The present study aimed to analyse the roles of cigarette smoke condensate (CSC) and nicotine on cell viability, cell migration/invasion and myofibroblastic differentiation using primary cultures of human gingival fibroblasts. MATERIAL AND METHODS: To compare the effects of CSC and nicotine, gingival fibroblasts were stimulated with CSC (0.4500 lg/mL) and the corresponding nicotine concentrations (0.02532 lg/mL) present in research cigarettes (1R3F). Cell viability was evaluated through the MTS assay. Cell migration and invasion were assessed through scratch wound assays, collagen nested matrices and trans well migration. a-Smooth muscle actin production was evaluated by western blotting. RESULTS: Cigarette smoke condensate at 50 lg/mL induced a moderate increase in cell viability, whereas the corresponding nicotine concentration (3.2 lg/mL) did not produce this response. Cigarette smoke condensate at 250 lg/mL, but not nicotine at 16 lg/mL (the corresponding nicotine concentration), induced cell death. Both nicotine and CSC stimulated cell migration (50 lg/mL CSC; 3.2 lg/mL nicotine). At 150 lg/mL, CSC inhibited cell migration; however, the corresponding concentration of nicotine (9.6 lg/mL), did not have this effect. Although both nicotine and CSC inhibited a-smooth muscle actin production, only the latter induced a statistically significant effect on this response. CONCLUSION: Cigarette smoke condensate may stimulate cell survival and migration at low concentrations and inhibit these cell responses at higher levels of exposure. Moreover, CSC may interfere in myofibroblastic differentiation.These results show that cigarette smoke, but not nicotine, may significantly alter cell viability, cell migration and myofibroblastic differentiation in gingival mesenchymal cells.
Assuntos
Gengiva/efeitos dos fármacos , Miofibroblastos/efeitos dos fármacos , Nicotiana , Nicotina/farmacologia , Agonistas Nicotínicos/farmacologia , Fumaça , Actinas/efeitos dos fármacos , Técnicas de Cultura de Células , Morte Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Corantes , Fibroblastos/efeitos dos fármacos , Gengiva/citologia , Humanos , Fumaça/efeitos adversos , Sais de Tetrazólio , Tiazóis , Nicotiana/efeitos adversosRESUMO
OBJECTIVES: Duplex ultrasound has become the reference standard in assessing the morphology and haemodynamics of the lower limb veins. The project described in this article was an initiative of the Union Internationale de Phlébologie (UIP). The aim was to obtain a consensus of international experts on the methodology and terminology to be used for assessment after treatment of incompetent superficial and perforating veins in the lower limb by ultrasound imaging. DESIGN: The study design was consensus meetings leading to a consensus document. METHODS: The UIP invited group submitted relevant literature references and written contributions concerning the methodology, terminology and value of duplex imaging after treatment. The authors prepared a draft document that was circulated to a larger group of experts and revised according to the comments received. Eventually, all participants agreed upon the final version of the article. RESULTS: Formal analysis of the results of interventions for varicose veins relies on adequate preoperative assessment and a careful description of the procedure employed. The timing of investigations of outcome should be classified as immediate (1-4 weeks), short-term (1 year), midterm (2-3 years) and long-term (5 years or more). The examination should employ standard methodology and formally described variables, which can be tailored to the intervention that was undertaken. The experts have made detailed recommendations concerning the methods to be used for duplex ultrasound examination and reporting after various treatments for varicose veins, including novel treatments under scientific study. CONCLUSIONS: Duplex ultrasonography is a fundamental component of the investigation of the lower limb venous system after treatment for varicose veins.
Assuntos
Extremidade Inferior/irrigação sanguínea , Ultrassonografia Doppler Dupla/normas , Varizes/terapia , Conferências de Consenso como Assunto , Medicina Baseada em Evidências , Hemodinâmica , Humanos , Valor Preditivo dos Testes , Fatores de Tempo , Resultado do Tratamento , Varizes/diagnóstico por imagem , Varizes/fisiopatologia , Veias/diagnóstico por imagem , Veias/fisiopatologiaRESUMO
Regulation of cell renewal in the periodontium is a critical cell function that has not been clarified. Sonic hedgehog (Shh) is a secreted signaling molecule that plays a key role during development and adult tissue homeostasis. In the present study, we have analyzed the role played by Shh in human periodontal ligament stem cell (HPLSC) proliferation. HPLSC were isolated with anti-STRO-1 antibodies. Shh increased the expression of GLI1 and PTC-1 and selectively stimulated cell proliferation in STRO-1(+) derived from adult periodontal ligament. Shh components were localized to primary cilia in STRO-1(+) cells after Shh stimulation. STRO-1(+) also expressed Shh, suggesting an autocrine-regulated phenomenon. Thus, we propose that Shh plays a critical role in the regulation of cell proliferation in STRO-1(+)/HPLSC.
Assuntos
Proteínas Hedgehog/farmacologia , Ligamento Periodontal/citologia , Células-Tronco/efeitos dos fármacos , Adolescente , Adulto , Antígenos de Superfície/análise , Comunicação Autócrina/fisiologia , Contagem de Células , Técnicas de Cultura de Células , Diferenciação Celular/fisiologia , Proliferação de Células/efeitos dos fármacos , Separação Celular , Cílios/ultraestrutura , Meios de Cultivo Condicionados , Humanos , Fatores de Transcrição Kruppel-Like/análise , Morfolinas/farmacologia , Proteínas Nucleares/análise , Receptores Patched , Ligamento Periodontal/efeitos dos fármacos , Purinas/farmacologia , Receptores de Superfície Celular/análise , Receptores Acoplados a Proteínas G/agonistas , Receptores Acoplados a Proteínas G/fisiologia , Receptor Smoothened , Fatores de Transcrição/análise , Adulto Jovem , Proteína GLI1 em Dedos de Zinco , Proteína Gli2 com Dedos de ZincoRESUMO
BACKGROUND AND OBJECTIVE: Statins have been used to control hypercholesterolemia. However, these drugs also exert pleiotropic effects that include the modulation of inflammation and cell signaling. The present study has analyzed the effects of simvastatin on several cell responses involved in tissue repair, including cell adhesion, cell migration and invasion, actin cytoskeleton remodeling and cell viability. MATERIAL AND METHODS: Primary cultures of gingival fibroblasts were stimulated with simvastatin. Cell adhesion was evaluated using a colorimetric assay. Cell spreading was evaluated microscopically. Cell migration and invasion were assessed using a scratch wound-healing assay and a bicameral cell culture system, respectively. Changes in actin cytoskeleton and focal adhesion assembly were evaluated through immunofluorescence for actin, vinculin and active ß1 integrin. Rac activation was evaluated by means of a pull-down assay. Cell viability was assessed using a colorimetric assay that determines mitochondrial functionality. Data analysis was performed using the Mann-Whitney U-test. RESULTS: Simvastatin diminished cell adhesion and spreading over a fibronectin matrix. It also altered the closure of scratch wounds induced on cell monolayers and cell invasion through a Transwell system. Simvastatin-treated cells displayed an altered lamellipodia with poorly developed focal adhesion contacts and reduced levels of ß1 integrin activation. During cell spreading, simvastatin diminished Rac activation. CONCLUSION: The present study shows that simvastatin may alter cell migration by disrupting the cell signaling networks that regulate the actin cytoskeleton dynamics. This mechanism may affect the response of gingival mesenchymal cells during wound healing.
Assuntos
Anticolesterolemiantes/farmacologia , Fibroblastos/efeitos dos fármacos , Gengiva/efeitos dos fármacos , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Sinvastatina/farmacologia , Actinas/análise , Adolescente , Adulto , Adesão Celular/efeitos dos fármacos , Técnicas de Cultura de Células , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Citoesqueleto/efeitos dos fármacos , Ativação Enzimática , Feminino , Imunofluorescência , Gengiva/citologia , Humanos , Integrina beta1/análise , Masculino , Pseudópodes/efeitos dos fármacos , Vinculina/análise , Adulto Jovem , Proteínas rac de Ligação ao GTP/análiseRESUMO
The lip vermillion constitutes a transition tissue, between oral mucosa and skin, where oral mucosal cells from epithelial and connective tissue compartments are exposed to ultraviolet (UV) sunlight. Fibroblasts are abundant resident cells of the connective tissue which are key regulators of extracellular matrix composition, as well as, epithelial and endothelial cell function. UVB light, an inherent component of sunlight, causes several alterations in skin fibroblasts, including premature senescence and increased cyclooxygenase (COX)-2 expression. To assess if UVB irradiation had similar effects on fibroblasts derived from human oral mucosa (HOM), primary cultures of HOM fibroblasts were irradiated with a single dose of 30 or 60 mJ/cm²of UVB light or sham-irradiated. Fibroblast proliferation was assessed from 3 to 48 hrs after UVB-irradiation utilizing [³H]-thymidine incorporation and MTT assays. In addition, COX-2 mRNA expression was detected by RT-PCR, and PGE2 production was assessed using enzyme immunoassay from 0.5 to 24 hrs after UVB-irradiation. The results showed a significant decrease in proliferation of UVB-irradiated HOM fibroblasts as compared to controls as measured by both [³H]-thymidine incorporation and MTT assays (p<0.001). HOM fibroblasts had increased COX-2 mRNA expression at 0.5 and 12 hrs after irradiation, and PGE2 production was elevated at 12 and 24 hrs post-irradiation as compared to controls (p<0.05). The results showed an inhibitory effect of a single dose of UVB irradiation on HOM fibroblast proliferation with an increase in COX-2 expression and activation. Therefore, photodamaged fibroblasts may play and important role in the pathogenesis of UV-induced lesions of the lip.
Assuntos
Humanos , /efeitos da radiação , Fibroblastos/efeitos da radiação , Mucosa Bucal/citologia , Raios Ultravioleta , Fibroblastos/enzimologia , Mucosa Bucal/efeitos da radiação , Mucosa Bucal/enzimologia , Proliferação de Células , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
BACKGROUND: The influence of the stromal microenvironment on the progression of epithelial cancers has been demonstrated. Unravelling the mechanisms by which stromal cells affect epithelial behaviour will contribute in understanding cellular malignancy. It has been proposed that redox environment has a role in the acquisition of malignancy. In this work, we studied the influence of epithelial cells on the stromal redox status and the consequence of this phenomenon on MCF-7 cell motility. METHODS: We analysed in a co-culture system, the effect of RMF-EG mammary stromal cells on the migratory capacity of MCF-7 cell line. To test whether the NOX-dependent stromal redox environment influences the epithelial migratory behaviour, we knocked down the expression of NOX4 using siRNA strategy. The effect of TGF-ß1 on NOX4 expression and activity was analysed by qPCR, and intracellular ROS production was measured by a fluorescent method. RESULTS: Migration of MCF-7 breast epithelial cells was stimulated when co-cultured with RMF-EG cells. This effect depends on stromal NOX4 expression that, in turn, is enhanced by epithelial soluble factors. Pre-treatment of stromal cells with TGF-ß1 enhanced this migratory stimulus by elevating NOX4 expression and intracellular ROS production. TGF-ß1 seems to be a major component of the epithelial soluble factors that stimulate NOX4 expression. CONCLUSIONS: Our results have identified that an increased stromal oxidative status, mainly provided by an elevated NOX4 expression, is a permissive element in the acquisition of epithelial migratory properties. The capacity of stromal cells to modify their intracellular ROS production, and accordingly, to increase epithelial motility, seems to depend on epithelial soluble factors among which TGF-ß1 have a decisive role.
Assuntos
Neoplasias da Mama/patologia , Mama/citologia , Movimento Celular , Células Epiteliais/fisiologia , NADPH Oxidases/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Células Estromais/metabolismo , Linhagem Celular , Linhagem Celular Tumoral , Técnicas de Cocultura , Fibroblastos , Humanos , NADPH Oxidase 4 , Células Estromais/fisiologia , Fator de Crescimento Transformador beta1/farmacologiaRESUMO
OBJECTIVES: Periodontal disease is characterized by an increased collagen metabolism. Although membrane type-1 matrix metalloproteinase (MT1-MMP) plays a critical role in collagen degradation, its involvement in human periodontitis remains to be determined. METHODS: MT1-MMP and TIMP-2 expression and distribution were evaluated in gingival tissue samples derived from 10 healthy and 12 periodontitis-affected human subjects. MT1-MMP and TIMP-2 expression were assessed through Western-blot of tissue homogenates. The main cell types involved in MT1-MMP and TIMP-2 production were evaluated by means of immunohistochemistry. RESULTS: Both MT1-MMP and TIMP-2 were significantly increased in periodontitis-affected gingival tissues when compared to healthy gingiva. Moreover, the balance between MT1-MMP and its inhibitor TIMP-2 was altered in periodontitis-affected tissues, suggesting an imbalance in this proteolytic axis. Immunohistochemistry demonstrated the expression of MT1-MMP in fibroblasts and macrophages in gingival tissues. MT1-MMP was detected in cells in close association with the gingival collagen matrix. TIMP-2 expression was identified in fibroblasts, macrophages and epithelial cells. CONCLUSIONS: Our observations show an increased expression of MT1-MMP and TIMP-2 in periodontitis-affected gingival tissues. The altered balance between these two molecular mediators of collagen remodeling suggests their involvement in human periodontal disease.
Assuntos
Gengiva/enzimologia , Metaloproteinase 14 da Matriz/metabolismo , Periodontite/enzimologia , Inibidor Tecidual de Metaloproteinase-2/metabolismo , Estudos de Casos e Controles , Gengiva/patologia , Humanos , Imuno-Histoquímica , Periodontite/patologia , Valores de ReferênciaRESUMO
BACKGROUND AND OBJECTIVES: Destruction of the supporting periodontal tissues is mediated by the action of several proteolytic enzymes. Urokinase is a serine protease that plays a key role in connective tissue destruction through conversion of plasminogen into plasmin. The present study was conducted to evaluate the effect of triclosan on the production and activity of urokinase in cultured gingival fibroblasts. MATERIAL AND METHODS: Urokinase production was studied in primary cultures of human gingival fibroblasts stimulated with tumor necrosis factor-alpha. Urokinase activity and production were evaluated using casein zymography and western blotting, respectively. Urokinase mRNA expression was evaluated using the reverse transcription-polymerase chain reaction. Triclosan was used to interfere with this stimulatory effect. The roles of different cell-signaling cascades involved in urokinase production were assessed through western blotting and immunofluorescence using several cell-signaling inhibitors. RESULTS: Tumor necrosis factor-alpha was found to be a strong stimulus for urokinase production and triclosan was able to inhibit this response at the protein and mRNA levels. Triclosan was also able to inhibit conversion of plasminogen into plasmin. Tumor necrosis factor-alpha-stimulated urokinase production was shown to be dependent on the nuclear factor-kappaB and c-Jun N-terminal kinase signaling pathways. Triclosan inhibited c-Jun N-terminal kinase phosphorylation and c-Jun production. CONCLUSIONS: Within the limits of this study, these results show that triclosan may inhibit urokinase production and plasminogen activation in gingival fibroblasts through modulation of the c-Jun N-terminal kinase signaling pathway.