Integrated molecular, biochemical, and physiological assessment unravels key extraction method mediated influences on rat neonatal cardiomyocytes

Jensen, Leonardo; Neri, Elida Adalgisa; Bassaneze, Vinícius; Oliveira, Nathalia Cavichiolli de; Dariolli, Rafael; Turaça, Lauro Thiago; Levy, Débora; Veronez, Douglas; Ferraz, Mariana Sacrini Ayres; Alencar, Adriano M.; Bydlowski, Sérgio Paulo; Cestari, Idágene Aparecida; Krieger, José Eduardo

doi:10.1002/jcp.26380

Cited by 15 publications

(18 citation statements)

References 57 publications

(66 reference statements)

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“…Fluorescence (F) was normalized to background intensity (F 0 ) during cell contraction and was plotted over the time course with ImageJ software. Calcium transients were calculated according to a reported method 49 . Ten successive F/F0 peaks in four selected zones per sample were used for calculating the parameter of time to peak, and the peaks in three time periods (5 s per time period) in four selected zones per sample were used to quantify the peak number within 5 s. Three independent samples per group were quantified.…”

Section: Methodsmentioning

confidence: 99%

Mussel-inspired conductive nanofibrous membranes repair myocardial infarction by enhancing cardiac function and revascularization

He¹,

Ye²,

Song³

et al. 2018

Theranostics

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The controversy between polypyrrole's (Ppy) biocompatibility and its aggregation on nanofibers impedes application of conductive Ppy-incorporated nanofibers to create engineered cardiac microenvironments. The purpose of this study was to fabricate a functional scaffold for engineering cardiac patches (ECP) using a high concentration of methyl acrylic anhydride-gelatin (GelMA)-Ppy nanoparticles, mussel-inspired crosslinker, and electrospun (ES)-GelMA/polycaprolactone (PCL) nanofibrous membrane.Methods: First, spherical GelMA-Ppy nanoparticles were obtained when the methacrylate groups of GelMA formed a self-crosslinked network through oxidative polymerization of Ppy. Second, GelMA-Ppy nanoparticles were uniformly crosslinked on the ES-GelMA/PCL membrane through mussel-inspired dopamine-N'N'-methylene-bis-acrylamide (dopamine-MBA) crosslinker. Finally, the feasibility of the dopa-based conductive functional ECP scaffold was investigated in vitro and in vivo.Results: The GelMA-Ppy nanoparticles displayed excellent biocompatibility at a high concentration of 50 mg/mL. The massive GelMA-Ppy nanoparticles could be uniformly distributed on the ES nanofibers through dopamine-MBA crosslinker without obvious aggregation. The high concentration of GelMA-Ppy nanoparticles produced high conductivity of the dopamine-based (dopa-based) conductive membrane, which enhanced the function of cardiomyocytes (CMs) and yielded their synchronous contraction. GelMA-Ppy nanoparticles could also modify the topography of the pristine ES-GelMA/PCL membrane to promote vascularization in vitro. Following transplantation of the conductive membrane-derived ECP on the infarcted heart for 4 weeks, the infarct area was decreased by about 50%, the left ventricular shortening fraction percent (LVFS%) was increased by about 20%, and the neovascular density in the infarct area was significantly increased by about 9 times compared with that in the MI group.Conclusion: Our study reported a facile and effective approach to developing a functional ECP that was based on a mussel-inspired conductive nanofibrous membrane. This functional ECP could repair infarct myocardium through enhancing cardiac function and revascularization.

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Section: Methodsmentioning

confidence: 99%

Mussel-inspired conductive nanofibrous membranes repair myocardial infarction by enhancing cardiac function and revascularization

He¹,

Ye²,

Song³

et al. 2018

Theranostics

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“…O comportamento do M R durante o ciclo de contração, para uma amostra de cardiomiócito WT pode ser visto na O ajuste da potência pela derivada do M R foi automatizado pelo nosso grupo de pesquisa e aplicada em um trabalho, visando investigar a influência de duas metodologias de extração de cardiomiócitos de ratos neonatos em suas propriedades mecânicas (Jensen et al, 2018)). A pesquisa foi realizada utilizando o TFM como metodologia experimental e, entre outras análises, a potência de contração e relaxamento foi utilizada para estudar as propriedades mecânicas dos cardiomiócitos em cada tipo de extração (maiores detalhes sobre esse trabalho podem ser vistos em (Jensen et al, 2018)).…”

Section: Potência De Contraçãounclassified

Desenvolvimento de ferramentas de análise da dinâmica de forças de tração durante o ciclo de contração de cardiomiócitos isolados >i<in vitro>/i<

Dorta¹

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The mechanical properties of the heart have been studied studied extensively in recent years, both at organ level, muscle tissue, and recently at the cellular level. Heart cells, or cardiomyocytes, are cell of the heart muscle, which is responsible for the permanent blood flow in the body, making up the atria and ventricles, chambers where blood enters and is pumped into the heart. The use of cardiomyocytes in scientific experiments has brought several experimental advantages, as well as the possibility of studying isolated cells from different areas of the heart. In the present work an analysis methodology was developed capable of measuring parameters related to the dynamics of traction forces os isolated cardiomyocytes during the contraction cycles, aiming to generate a better understanding in the area of cardiac regeneration. For this we used two cardiomyocytes line, WT, which are from healthy cardiomyocytes, an KO-CRP3, which are from cardiomyocytes that have malfunction in cardiac muscle architecture. The developed methodology aims to analyze parameters such as the pulse rate of the cardiomyocytes, the alignment of the traction forces during the contraction cycles and the power performed by the cardiomyocytes during the contraction and relaxation. These parameters were analyzed under two conditions, with cardiomyocytes at basal state, and stimulated by a drug, isoproterenol. Thus it was possible to analyze the mechanical response of cardiomyocytes to the drug used, and the difference in the behavior of teh parameters between the cardiomyocytes line.

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“…Geralmente, proveniente de animais como camundongos ou ratos, devido a sua relativa robustez em cultura e custo [10]. Além da existência de vários protocolos consolidados para o isolamento desse tipo de célula [28], [29]. Quando isolados, os cardiomiócitos neonatais podem durar varias semanas em cultura.…”

Section: Mecânica Celular Dos Cardiomiócitosunclassified

“…Por esse motivo, utilizamos esse tipo de célula com modelo in vitro para o estudo das suas propriedades mecânica e teste com fármaco.O tipo de cardiomiócitos neonatais utilizados em especifico no trabalho, são derivados de ratos (Rattus norvegicus) da linhagem Wistar. São extraídos do coração de recémnascidos após 1 dia de vida, utilizado o método pre-plating[28].…”

unclassified

Estudo das propriedades mecânicas da contração executada por células cardíacas isoladas aderidas em superfície mimetizada

Querino¹

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Recent evidence demonstrates that the study of cellular mechanical behavior is crucial for understanding various physiological processes such as differentiation, migration and contraction. Both the cellular tensile forces and the extracellular matrix (ECM) mechanical properties contribute to the biological regulation of the cell, which was previously understood based on the principles of biochemistry. The importance of mechanical forces in cellular signaling in different processes is evident. Our work has as main objective the study of the mechanical properties of cyclic contraction, performed by isolated cardiac cells, adhered to a mimic surface (mimics the tissue). Motivated by the demand for new methods of investigation of cardiomyocytes in vitro. Where the approach focuses on the study of mechanical properties. We analyzed and quantified the mechanical response signal of these cells when adhered, aiming to characterize the mechanical response of these cells to the development of new methodologies aimed at the use of these cells with in vitro models, evaluating possible effects that manifest on organ based on mechanical parameters. In this work we used as samples, neonatal rats cardiomyocytes in basal state and with chemical stimulation using isoproterenol. For this, we used the modified Force and Traction Microscopy (TFM) technique to quantify the contraction in time, and signal analysis and distribution methods, which allow the characterization of the mechanical signal of cyclic contraction between isoproterenol-stimulated basal state. Among the results reported in this work is the characterization of the contractile activity regions, characterized by their discrete distribution along the cell edge, presenting a multipolar stress field with anisotropy. In addition to its distribution and guidance on the area of adhesion. Another finding finding is the shape and steps of the contractile cycle of the voltage signal and voltage variation, showing their similarity to the inverted Van Der Pol oscillation model. Finally, we report the variation of the increase in the cellular heartbeat frequency as a function of the basal rate. In addition, we show the conditions of cell adhesion area and basal rate for the maximum increase in heart cell frequency. The vi results presented in this Thesis have applications in basic and clinical research.

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Integrated molecular, biochemical, and physiological assessment unravels key extraction method mediated influences on rat neonatal cardiomyocytes

Cited by 15 publications

References 57 publications

Mussel-inspired conductive nanofibrous membranes repair myocardial infarction by enhancing cardiac function and revascularization

Mussel-inspired conductive nanofibrous membranes repair myocardial infarction by enhancing cardiac function and revascularization

Desenvolvimento de ferramentas de análise da dinâmica de forças de tração durante o ciclo de contração de cardiomiócitos isolados >i<in vitro>/i<

Estudo das propriedades mecânicas da contração executada por células cardíacas isoladas aderidas em superfície mimetizada

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