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1.
Cell Biochem Biophys ; 2024 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-38498099

RESUMO

In vitro cellular models provide valuable insights into the adaptive biochemical mechanisms triggered by cells to cope with the stress situation induced by hypoxia and reoxygenation cycles. The first biological data generated in studies based on this micrometric life-scale has the potential to provide us a global overview about the main biochemical phenomena presented in some reported preconditioning therapies in life-scale of higher dimensions. Thus, in this study, a cell incubator was designed and manufactured to produce a cellular model of heart hypoxia followed by reoxygenation (HfR) through consecutive repetitions of hypoxia-normoxia gas exchange. Samples of cellular extracts and culture media were obtained from non-proliferative cardiomyocytes (CMs) cultivated under challenging HfR (stressed CMs) and regular cultivation (unstressed CMs) in rounds of four days for each case. Metabolomic based on proton magnetic resonance spectroscopy (1H-MRS) was used as an analytical approach to identify and quantify the metabolomes of these samples, the endo- and exo-metabolome. Despite the stressed CMs presented over 90% higher cellular death rate compared to the unstressed CMs, the metabolic profiles indicates that the surviving cells up-regulate their amino acid metabolism either by active protein degradation or by the consumption of culture media components to increase coenzyme A-dependent metabolic pathways. This cell auto-regulation mechanism could be well characterized in the first two days when the difference smears off under once the metabolomes become similar. The metabolic adaptations of stressed CMs identified the relevance of the cyclic oxidation/reduction reactions of nicotinamide adenine dinucleotide phosphate molecules, NADP+/NADPH, and the increased tricarboxylic acid cycle activity in an environment overloaded with such a powerful antioxidant agent to survive an extreme HfR challenge. Thus, the combination of cellular models based on CMs, investigative methods, such as metabolomic and 1H-MRS, and the instrumental development of hypoxia incubator shown in this work were able to provide the first biochemical evidences behind therapies of gaseous exchanges paving the way to future assays.

2.
J Fluoresc ; 2024 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-38183590

RESUMO

The interaction between silver nanoparticles (AgNPs) and molecules producing coronas plays a key role in cytotoxicity mechanisms. Once adsorbed coronas determine the destiny of nanomaterials in vivo, their effective deployment in the biomedical field requires a comprehensive understanding of the dynamic interactions of biomolecules with nanoparticles. In this work, we characterized 40 nm AgNPs in three different nutritional cell media at different molar concentrations and incubation times to study the binding mechanism of molecules on surface nanoparticles. In addition, their cytotoxic effects have been studied in three cell lineages used as tissue regeneration models: FN1, HUV-EC-C, RAW 264.7. According to the data, when biomolecules from DMEM medium were in contact with AgNPs, agglomeration and precipitation occurred. However, FBS medium proteins indicated the formation of coronas over the nanoparticles. Nonetheless, little adsorption of molecules around the nanoparticles was observed when compared to DMEM supplemented with 10% FBS. These findings indicate that when nanoparticles and bioproteins from supplemented media interact, inorganic salts from DMEM contribute to produce large bio-coronas, the size of which varies with the concentration and time. The static quenching mechanism was shown to be responsible for the fluorescence quenching of the bioprotein aggregates on the AgNPs surface. The calculated bioprotein-nanoparticle surface binding constants were on the order of 105 M-1 at 37 °C, with hydrophobic interactions driven by enthalpy and entropy playing a role, as confirmed by thermodynamic analysis. Cytotoxicity data showed a systematic degrowth in the viable cell population as the number of nanoparticles increased and the diameter of coronas decreased. Cytotoxic intervals associated with half decrease of cell population were established for AgNPs molar concentration of 75 µM for 24 h and 50 µM for 48 h. In summary, through the cytotoxicity mechanism of bio-coronas we are able to manipulate cells' expansion rates to promote specific processes, such inflammatory mechanisms, at different time instants.

3.
In Vitro Cell Dev Biol Anim ; 56(8): 604-613, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32914385

RESUMO

Knockout of multifunction gene cysteine- and glycine-rich protein 3 (CSRP3) in cardiomyocytes (CMs) of mice leads to heart dilation, severely affecting its functions. In humans, CSRP3 mutations are associated with hypertrophic (HCM) and dilated cardiomyopathy (DCM). The absence of the CSRP3 expression produces unknown effects on in vitro neonatal CMs' metabolism. The metabolome changes in culture media conditioned by CSRP3 knockout (KO-CSRP3), and wild type (WT) neonatal cardiomyocytes were investigated under untreated or after metabolic challenging conditions produced by isoproterenol (ISO) stimulation, by in vitro high-resolution proton magnetic resonance spectroscopy (1H-MRS)-based metabolomics. Metabolic differences between neonatal KO-CSRP3 and WT rats' CMs were identified. After 72 h of culture, ISO administration was associated with increased CMs' energy requirements and increased levels of threonine, alanine, and 3-hydroxybutyrate in both neonatal KO-CSRP3 and WT CMs conditioned media. When compared with KO-CSRP3, culture media derived from WT cells presented higher lactate concentrations either under basal or ISO-stimulated conditions. The higher activity of ketogenic biochemical pathways met the elevated energy requirements of the contractile cells. Both cells are considered phenotypically indistinguishable in the neonatal period of animal lives, but the observed metabolic stress responses of KO-CSRP3 and WT CMs to ISO were different. KO-CSRP3 CMs produced less lactate than WT CMs in both basal and stimulated conditions. Mainly, ISO-stimulated conditions produced evidence for lactate overload within KO-CSRP3 CMs, while WT CMs succeeded to manage the metabolic stress. Thus, 1H-MRS-based metabolomics was suitable to identify early inefficient energetic metabolism in neonatal KO-CSRP3 CMs. These results may reflect an apparent lower lactate transport and consumption, in association with protein catabolism.


Assuntos
Meios de Cultura/química , Proteínas com Domínio LIM/metabolismo , Proteínas Musculares/metabolismo , Miócitos Cardíacos/metabolismo , Espectroscopia de Prótons por Ressonância Magnética , Animais , Animais Recém-Nascidos , Forma Celular , Análise Discriminante , Isoproterenol/farmacologia , Proteínas com Domínio LIM/deficiência , Análise dos Mínimos Quadrados , Proteínas Musculares/deficiência , Miócitos Cardíacos/citologia , Miócitos Cardíacos/efeitos dos fármacos , Ratos , Estatística como Assunto
4.
J Phys Chem B ; 124(36): 7842-7848, 2020 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-32790314

RESUMO

We present a new Monte Carlo method to simulate ionic liquids in slab geometry at constant potential. The algorithm is built upon two previous methods while retaining the advantages of each of them. The method is tested against a Poisson-Boltzmann theory and the constant surface charge ensemble, achieving consistency among all of them. We then analyze the computational time of the developed algorithm, showing substantial speedup in relation to the method of Kiyohara and Asaka [J. Chem. Phys., 2007, 126, 214704]. As an application of our method, we investigate crowding and overscreening in confined room-temperature ionic liquids. We show that we can switch between two behaviors of the double layer by changing the Bjerrum length alone.

5.
Auris Nasus Larynx ; 47(1): 98-104, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31272842

RESUMO

OBJECTIVE: Chronic Rhinosinusitis with Nasal Polyps (CRSwNP) is a disease that features a mechanical dysfunction involving chronic inflammation and altered tissue remodeling. In this study, we aim to evaluate the fibroblast morphology and its cellular traction force in primary fibroblasts cell cultures obtained from both healthy individuals (n=7) and patients with CRSwNP (n=8). METHODS: Using a Traction-force Microscopy we analyzed parameters of Force/Tension in fibroblasts cultures in both experimental groups. RESULTS: The analysis of the Projected Area of Cell revealed that fibroblasts derived from nasal mucosa of healthy individuals have an area on average 39.24% larger than the fibroblasts obtained from the nasal polyp tissue. We also observed that the parameters directly related to the force of the cell, Max Cumulative Force and Net Contractile Moment, presented a high Force/Tension per unit of area in the fibroblasts derived from the healthy nasal mucosa (on average 41% and 52.54% higher than the fibroblasts of the nasal polyp respectively). CONCLUSION: Our results demonstrate a cellular mechanism that may be associated with the mechanical dysfunction found in the Nasal Polyp tissue. The weak traction force of nasal polyp-derived fibroblast may, in lower dimensions, impact on the remodeling of nasal mucosa in CRSwNP.


Assuntos
Fenômenos Biomecânicos , Fibroblastos/ultraestrutura , Pólipos Nasais/ultraestrutura , Pseudópodes/ultraestrutura , Estudos de Casos e Controles , Doença Crônica , Feminino , Fibroblastos/patologia , Fibroblastos/fisiologia , Humanos , Masculino , Microscopia de Força Atômica , Microscopia de Contraste de Fase , Pessoa de Meia-Idade , Pólipos Nasais/patologia , Pólipos Nasais/fisiopatologia , Cultura Primária de Células , Pseudópodes/patologia , Rinite/patologia , Sinusite/patologia
6.
Int Arch Otorhinolaryngol ; 23(2): 241-249, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30956711

RESUMO

Introduction The importance of our study lies in the fact that we have demonstrated the occurrence of mechanical dysfunction within polypoid tissues, which promotes the development of polyps in the nasal cavity. Objective To change the paradigm of nasal polyposis (NP). In this new conception, the chronic nasal inflammatory process that occurs in response to allergies, to pollution, to changes in the epithelial barrier, or to other factors is merely the trigger of the development of the disease in individuals with a genetic predisposition to an abnormal tissue remodeling process, which leads to a derangement of the mechanical properties of the nasal mucosa and, consequently, allows it to grow unchecked. Data Synthesis We propose a fundamentally new approach to intervening in the pathological process of NP, addressing biomechanical properties, fluid dynamics, and the concept of surface tension. Conclusion The incorporation of biomechanical knowledge into our understanding of NP provides a new perspective to help elucidate the physiology and the pathology of nasal polyps, and new avenues for the treatment and cure of NP.

7.
Int. arch. otorhinolaryngol. (Impr.) ; 23(2): 241-249, 2019. ilus, tab
Artigo em Inglês | LILACS | ID: biblio-1015650

RESUMO

Introduction: The importance of our study lies in the fact that we have demonstrated the occurrence ofmechanical dysfunction within polypoid tissues, which promotes the development of polyps in the nasal cavity. Objective: To change the paradigm of nasal polyposis (NP). In this new conception, the chronic nasal inflammatory process that occurs in response to allergies, to pollution, to changes in the epithelial barrier, or to other factors is merely the trigger of the development of the disease in individuals with a genetic predisposition to an abnormal tissue remodeling process, which leads to a derangement of the mechanical properties of the nasal mucosa and, consequently, allows it to grow unchecked. Data: Synthesis We propose a fundamentally new approach to intervening in the pathological process of NP, addressing biomechanical properties, fluid dynamics, and the concept of surface tension. Conclusion: The incorporation of biomechanical knowledge into our understanding of NP provides a new perspective to help elucidate the physiology and the pathology of nasal polyps, and new avenues for the treatment and cure of NP (AU)


Assuntos
Humanos , Pólipos Nasais/fisiopatologia , Pólipos Nasais/patologia , Inflamação/fisiopatologia , Sinusite/fisiopatologia , Fenômenos Biomecânicos , Brasil , Mecânica dos Fluidos , Doença Crônica , Edema/fisiopatologia , Matriz Extracelular/patologia , Pressão Hidrostática , Mucosa Nasal/fisiopatologia , Mucosa Nasal/patologia
8.
Soft Matter ; 12(41): 8506-8511, 2016 Oct 19.
Artigo em Inglês | MEDLINE | ID: mdl-27722665

RESUMO

The cytoskeleton (CSK) is a tensed fiber framework that supports, shapes and stabilizes the cell. The CSK is in a constant state of remodeling, moreover, which is an active non-equilibrium thermodynamic process. We report here that cytoskeletal remodeling involves reconfigurations that are not only sudden but also are transmitted to great distances within the cell in a fashion reminiscent of quakes in the Earth's crust. Remarkably, these events in the cell conform both qualitatively and quantitatively to empirical laws typical of earthquakes, including hierarchical fault structures, cumulative energy distributions following the Gutenberg-Richter law, and rate of after-shocks following Omori's law. While it is well-established that remodeling and stabilization of the cytoskeleton are non-equilibrium process, these new unanticipated observations establish that these processes are also remarkably non-local and strongly cooperative.


Assuntos
Citoesqueleto/fisiologia , Miócitos de Músculo Liso/citologia , Células Cultivadas , Humanos , Termodinâmica
9.
Environ Sci Pollut Res Int ; 23(10): 9862-70, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26856867

RESUMO

Diesel exhaust particles (DEPs) from diesel engines produce adverse alterations in cells of the airways by activating intracellular signaling pathways and apoptotic gene overexpression, and also by influencing metabolism and cytoskeleton changes. This study used human bronchial epithelium cells (BEAS-2B) in culture and evaluates their exposure to DEPs (15ug/mL for 1 and 2 h) in order to determine changes to cell rheology (viscoelasticity) and gene expression of the enzymes involved in oxidative stress, apoptosis, and cytotoxicity. BEAS-2B cells exposed to DEPs were found to have a significant loss in stiffness, membrane stability, and mitochondrial activity. The genes involved in apoptosis [B cell lymphoma 2 (BCL-2 and caspase-3)] presented inversely proportional expressions (p = 0.05, p = 0.01, respectively), low expression of the genes involved in antioxidant responses [SOD1 (superoxide dismutase 1); SOD2 (superoxide dismutase 2), and GPx (glutathione peroxidase) (p = 0.01)], along with an increase in cytochrome P450, family 1, subfamily A, polypeptide 1 (CYP1A1) (p = 0.01). These results suggest that alterations in cell rheology and cytotoxicity could be associated with oxidative stress and imbalance between pro- and anti-apoptotic genes.


Assuntos
Antioxidantes/metabolismo , Apoptose/efeitos dos fármacos , Brônquios/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Material Particulado/toxicidade , Emissões de Veículos/toxicidade , Apoptose/genética , Brônquios/metabolismo , Brônquios/patologia , Caspase 3/genética , Caspase 3/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Glutationa Peroxidase/genética , Glutationa Peroxidase/metabolismo , Humanos , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Tamanho da Partícula , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Reologia
10.
PLoS One ; 10(12): e0145062, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26661469

RESUMO

AIMS: Recent evidence shows the rigidity of vascular smooth muscle cells (VSMC) contributes to vascular mechanics. Arterial rigidity is an independent cardiovascular risk factor whose associated modifications in VSMC viscoelasticity have never been investigated. This study's objective was to evaluate if the arterial rigidity risk factors aging, African ancestry, female sex, smoking and diabetes mellitus are associated with VMSC stiffening in an experimental model using a human derived vascular smooth muscle primary cell line repository. METHODS: Eighty patients subjected to coronary artery bypass surgery were enrolled. VSMCs were extracted from internal thoracic artery fragments and mechanically evaluated using Optical Magnetic Twisting Cytometry assay. The obtained mechanical variables were correlated with the clinical variables: age, gender, African ancestry, smoking and diabetes mellitus. RESULTS: The mechanical variables Gr, G'r and G"r had a normal distribution, demonstrating an inter-individual variability of VSMC viscoelasticity, which has never been reported before. Female sex and smoking were independently associated with VSMC stiffening: Gr (apparent cell stiffness) p = 0.022 and p = 0.018, R2 0.164; G'r (elastic modulus) p = 0.019 and p = 0.009, R2 0.184 and G"r (dissipative modulus) p = 0.011 and p = 0.66, R2 0.141. CONCLUSION: Female sex and smoking are independent predictors of VSMC stiffening. This pro-rigidity effect represents an important element for understanding the vascular rigidity observed in post-menopausal females and smokers, as well as a potential therapeutic target to be explored in the future. There is a significant inter-individual variation of VSMC viscoelasticity, which is slightly modulated by clinical variables and probably relies on molecular factors.


Assuntos
Músculo Liso Vascular/fisiologia , Fumar , Fatores Etários , Idoso , População Negra , Doenças Cardiovasculares/patologia , Doenças Cardiovasculares/cirurgia , Células Cultivadas , Ponte de Artéria Coronária , Diabetes Mellitus Tipo 2/patologia , Módulo de Elasticidade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Músculo Liso Vascular/citologia , Pós-Menopausa , Fatores de Risco , Fatores Sexuais
11.
Res. Biomed. Eng. (Online) ; 31(4): 328-333, Oct.-Dec. 2015. graf
Artigo em Inglês | LILACS | ID: biblio-829452

RESUMO

Introduction: One of the fundamental structural elements of the cell is the cytoskeleton. Along with myosin, actin microfilaments are responsible for cellular contractions, and their organization may be related to pathological changes in myocardial tissue. Due to the complexity of factors involved, numerical modeling of the cytoskeleton has the potential to contribute to a better understanding of mechanical cues in cellular activities. In this work, a systematic method was developed for the reconstruction of an actomyosin topology based on the displacement exerted by the cell on a flexible substrate. It is an inverse problem which could be considered a phenomenological approach to traction force microscopy (TFM). Methods An actomyosin distribution was found with a topology optimization method (TOM), varying the material density and angle of contraction of each element of the actomyosin domain. The routine was implemented with a linear material model for the bidimensional actomyosin elements and tridimensional substrate. The topology generated minimizes the nodal displacement squared differences between the generated topology and experimental displacement fields obtained by TFM. The structure resulting from TOM was compared to the actin structures observed experimentally with a GFP-attached actin marker. Results The optimized topology reproduced the main features of the experimental actin and its squared displacement differences were 11.24 µm2, 27.5% of the sum of experimental squared nodal displacements (40.87 µm2). Conclusion This approach extends the literature with a model for the actomyosin structure capable of distributing anisotropic material freely, allowing heterogeneous contraction over the cell extension.

13.
PLoS One ; 10(5): e0126440, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25946314

RESUMO

A cell mechanical stimulation equipment, based on cell substrate deformation, and a more sensitive method for measuring adhesion of cells were developed. A probe, precisely positioned close to the cell, was capable of a vertical localized mechanical stimulation with a temporal frequency of 207 Hz, and strain magnitude of 50%. This setup was characterized and used to probe the response of Human Umbilical Endothelial Vein Cells (HUVECs) in terms of calcium signaling. The intracellular calcium ion concentration was measured by the genetically encoded Cameleon biosensor, with the Transient Receptor Potential cation channel, subfamily M, member 7 (TRPM7) expression inhibited. As TRPM7 expression also regulates adhesion, a relatively simple method for measuring adhesion of cells was also developed, tested and used to study the effect of adhesion alone. Three adhesion conditions of HUVECs on polyacrylamide gel dishes were compared. In the first condition, the substrate is fully treated with Sulfo-SANPAH crosslinking and fibronectin. The other two conditions had increasingly reduced adhesion: partially treated (only coated with fibronectin, with no use of Sulfo-SANPAH, at 5% of the normal amount) and non-treated polyacrylamide gels. The cells showed adhesion and calcium response to the mechanical stimulation correlated to the degree of gel treatment: highest for fully treated gels and lowest for non-treated ones. TRPM7 inhibition by siRNA on HUVECs caused an increase in adhesion relative to control (no siRNA treatment) and non-targeting siRNA, but a decrease to 80% of calcium response relative to non-targeting siRNA which confirms the important role of TRPM7 in mechanotransduction despite the increase in adhesion.


Assuntos
Sinalização do Cálcio/fisiologia , Adesão Celular/fisiologia , Mecanotransdução Celular/fisiologia , Proteínas Serina-Treonina Quinases/genética , Canais de Cátion TRPM/genética , Técnicas Biossensoriais , Cálcio/metabolismo , Sinalização do Cálcio/genética , Adesão Celular/genética , Células Cultivadas , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Fenômenos Mecânicos , Proteínas Serina-Treonina Quinases/metabolismo , Interferência de RNA , RNA Interferente Pequeno , Canais de Cátion TRPM/metabolismo
14.
Environ Toxicol ; 30(11): 1297-308, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24777914

RESUMO

Particulate matter from diesel exhaust (DEP) has toxic properties and can activate intracellular signaling pathways and induce metabolic changes. This study was conducted to evaluate the activation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) and to analyze the mucin profile (acid (AB(+) ), neutral (PAS(+) ), or mixed (AB/PAS(+) ) mucus) and vacuolization (V) of tracheal explants after treatment with 50 or 100 µg/mL DEP for 30 or 60 min. Western blot analyses showed small increases in ERK1/2 and JNK phosphorylation after 30 min of 100 µg/mL DEP treatment compared with the control. An increase in JNK phosphorylation was observed after 60 min of treatment with 50 µg/mL DEP compared with the control. We did not observe any change in the level of ERK1/2 phosphorylation after treatment with 50 µg/mL DEP. Other groups of tracheas were subjected to histological sectioning and stained with periodic acid-Schiff (PAS) reagent and Alcian Blue (AB). The stained tissue sections were then subjected to morphometric analysis. The results obtained were compared using ANOVA. Treatment with 50 µg/mL DEP for 30 min or 60 min showed a significant increase (p < 0.001) in the amount of acid mucus, a reduction in neutral mucus, a significant reduction in mixed mucus, and greater vacuolization. Our results suggest that compounds found in DEPs are able to activate acid mucus production and enhance vacuolization and cell signaling pathways, which can lead to airway diseases.


Assuntos
Poluentes Atmosféricos/toxicidade , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Mucinas/metabolismo , Material Particulado/toxicidade , Traqueia/efeitos dos fármacos , Emissões de Veículos/toxicidade , Animais , Apoptose/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/patologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Muco/metabolismo , Fosforilação , Transdução de Sinais/efeitos dos fármacos , Traqueia/metabolismo , Traqueia/patologia
15.
Arq Bras Cardiol ; 102(4): 403-12, 2014 Apr.
Artigo em Inglês, Português | MEDLINE | ID: mdl-24844877

RESUMO

Natural myocardial markers, or speckles, originated from constructive and destructive interference of ultrasound in the tissues may provide early diagnosis of myocardial changes and be used in the prediction of some cardiac events. Due to its relatively temporal stability, speckles can be tracked by dedicated software along the cardiac cycle, enabling the analysis of the systolic and diastolic function. They are identified by either conventional 2D grey scale and by 3D echo, conferring independence of the insonation angle, thus allowing assessment of cardiac mechanics in the three spatial planes: longitudinal, circumferential, and radial. The purposes of the present paper are: to discuss the role and the meaning of cardiac strain obtained by speckle tracking during the evaluation of cardiac physiology and to discuss clinical applications of this novel echocardiographic technology.


Assuntos
Ecocardiografia/métodos , Cardiopatias/diagnóstico por imagem , Coração/fisiologia , Cardiopatias/fisiopatologia , Humanos , Ilustração Médica , Valor Preditivo dos Testes , Valores de Referência , Reprodutibilidade dos Testes , Função Ventricular/fisiologia
16.
Arq. bras. cardiol ; 102(4): 403-412, abr. 2014. tab, graf
Artigo em Português | LILACS | ID: lil-709315

RESUMO

Speckles, ou marcadores naturais do miocárdio, originam se da interferência construtiva e destrutiva do feixe de ultrassom que incide sobre os tecidos, podem fornecer um diagnóstico precoce das alterações miocárdicas e atuar na predição de certos eventos cardíacos. Devido à sua relativa estabilidade temporal, os speckles podem ser rastreados durante o ciclo cardíaco por software dedicados, promovendo a análise da função sistólica e diastólica. São identificados tanto pela escala de cinza da ecocardiografia 2D convencional quanto pela ecocardiografia 3D, sendo independentes do ângulo de incidência do ultrassom, permitindo assim a avaliação da mecânica cardíaca nos três planos espaciais: longitudinal, circunferencial e radial. O objetivo do presente artigo é discutir o papel e o significado da deformação cardíaca obtida por meio do speckle tracking durante a avaliação da fisiologia cardíaca, e discutir as aplicações clínicas desta tecnologia ecocardiográfica inovadora.


Natural myocardial markers, or speckles, originated from constructive and destructive interference of ultrasound in the tissues may provide early diagnosis of myocardial changes and be used in the prediction of some cardiac events. Due to its relatively temporal stability, speckles can be tracked by dedicated software along the cardiac cycle, enabling the analysis of the systolic and diastolic function. They are identified by either conventional 2D grey scale and by 3D echo, conferring independence of the insonation angle, thus allowing assessment of cardiac mechanics in the three spatial planes: longitudinal, circumferential, and radial. The purposes of the present paper are: to discuss the role and the meaning of cardiac strain obtained by speckle tracking during the evaluation of cardiac physiology and to discuss clinical applications of this novel echocardiographic technology.


Assuntos
Humanos , Ecocardiografia/métodos , Cardiopatias , Coração/fisiologia , Cardiopatias/fisiopatologia , Ilustração Médica , Valor Preditivo dos Testes , Valores de Referência , Reprodutibilidade dos Testes , Função Ventricular/fisiologia
17.
Am J Physiol Heart Circ Physiol ; 306(4): H505-16, 2014 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-24337458

RESUMO

Vascular smooth muscle cells (VSMCs) are thought to assume a quiescent and homogeneous mechanical behavior after arterial tree development phase. However, VSMCs are known to be molecularly heterogeneous in other aspects and their mechanics may play a role in pathological situations. Our aim was to evaluate VSMCs from different arterial beds in terms of mechanics and proteomics, as well as investigate factors that may influence this phenotype. VSMCs obtained from seven arteries were studied using optical magnetic twisting cytometry (both in static state and after stretching) and shotgun proteomics. VSMC mechanical data were correlated with anatomical parameters and ultrastructural images of their vessels of origin. Femoral, renal, abdominal aorta, carotid, mammary, and thoracic aorta exhibited descending order of stiffness (G, P < 0.001). VSMC mechanical data correlated with the vessel percentage of elastin and amount of surrounding extracellular matrix (ECM), which decreased with the distance from the heart. After 48 h of stretching simulating regional blood flow of elastic arteries, VSMCs exhibited a reduction in basal rigidity. VSMCs from the thoracic aorta expressed a significantly higher amount of proteins related to cytoskeleton structure and organization vs. VSMCs from the femoral artery. VSMCs are heterogeneous in terms of mechanical properties and expression/organization of cytoskeleton proteins along the arterial tree. The mechanical phenotype correlates with the composition of ECM and can be modulated by cyclic stretching imposed on VSMCs by blood flow circumferential stress.


Assuntos
Artérias/fisiologia , Músculo Liso Vascular/fisiologia , Miócitos de Músculo Liso/fisiologia , Animais , Artérias/metabolismo , Ciclo Celular/fisiologia , Colágeno/metabolismo , Elastina/metabolismo , Feminino , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , Proteômica , Sus scrofa
18.
Biorheology ; 49(5-6): 365-73, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23380902

RESUMO

One drawback of in vitro cell culturing is the dedifferentiation process that cells experience. Smooth muscle cells (SMC) also change molecularly and morphologically with long term culture. The main objective of this study was to evaluate if culture passages interfere in vascular SMC mechanical behavior. SMC were obtained from five different porcine arterial beds. Optical magnetic twisting cytometry (OMTC) was used to characterize mechanically vascular SMC from different cultures in distinct passages and confocal microscopy/western blotting, to evaluate cytoskeleton and extracellular matrix proteins. We found that vascular SMC rigidity or viscoelastic complex modulus (G) decreases with progression of passages. A statistically significant negative correlation between G and passage was found in four of our five cultures studied. Phalloidin-stained SMC from higher passages exhibited lower mean signal intensity per cell (confocal microscopy) and quantitative western blotting analysis showed a decrease in collagen I content throughout passages. We concluded that vascular SMC progressively lose their stiffness with serial culture passaging. Thus, limiting the number of passages is essential for any experiment measuring viscoelastic properties of SMC in culture.


Assuntos
Músculo Liso Vascular/citologia , Miócitos de Músculo Liso/fisiologia , Animais , Células Cultivadas , Colágeno Tipo I/metabolismo , Módulo de Elasticidade , Magnetismo , Microscopia Confocal , Miócitos de Músculo Liso/citologia , Suínos
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