Name: Elastômeros magneto-reológicos micropadronizados para impulsionar mudanças no alinhamento de cardiomiócitos (Vídeo) | JoVE Journal
Uploaded: 2025-06-10T12:40:00.000+00:00
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Description: Aprenda a fabricar elastômeros magneto-reológicos micropadronizados para estudar o alinhamento de cardiomiócitos em resposta a mudanças dinâmicas de rigidez. Esta plataforma auxilia pesquisadores em cardiologia.

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<li>Jaalouk, D. E., Lammerding, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mechanotransduction+gone+awry%5BTitle%5D)+AND+%22Nat+Rev+Mol+Cell+Biol%22%5BJournal%5D)">Mechanotransduction gone awry.</a> Nat Rev Mol Cell Biol. 10 (1), 63-73 (2009).</li>
<li>Vite, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Extracellular+stiffness+induces+contractile+dysfunction+in+adult+cardiomyocytes+via+cellautonomous+and+microtubuledependent+mechanisms%5BTitle%5D)+AND+%22Basic+Res+Cardiol%22%5BJournal%5D)">Extracellular stiffness induces contractile dysfunction in adult cardiomyocytes via cellautonomous and microtubuledependent mechanisms.</a> Basic Res Cardiol. 117 (1), 41(2023).</li>
<li>Stowers, R. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Matrix+stiffness+induces+a+tumorigenic+phenotype+in+mammary+epithelium+through+changes+in+chromatin+accessibility%5BTitle%5D)+AND+%22Nat+Biomed+Eng%22%5BJournal%5D)">Matrix stiffness induces a tumorigenic phenotype in mammary epithelium through changes in chromatin accessibility.</a> Nat Biomed Eng. 3 (12), 1009-1019 (2019).</li>
<li>Engler, A. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Embryonic+cardiomyocytes+beat+best+on+a+matrix+with+heartlike+elasticity%3A+scarlike+rigidity+inhibits+beating%5BTitle%5D)+AND+%22J+Cell+Sci%22%5BJournal%5D)">Embryonic cardiomyocytes beat best on a matrix with heartlike elasticity: scarlike rigidity inhibits beating.</a> J Cell Sci. 121 (22), 3794-3802 (2008).</li>
<li>Rodriguez, A. G., Han, S. J., Regnier, M., Sniadecki, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Substrate+stiffness+increases+twitch+power+of+neonatal+cardiomyocytes+in+correlation+with+changes+in+myofibril+structure+and+intracellular+calcium%5BTitle%5D)+AND+%22Biophys+J%22%5BJournal%5D)">Substrate stiffness increases twitch power of neonatal cardiomyocytes in correlation with changes in myofibril structure and intracellular calcium.</a> Biophys J. 101 (10), 2455-2464 (2011).</li>
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<li>Clark, P., Connolly, P., Curtis, A. S. G., Dow, J. A., Wilkinson, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Topographical+control+of+cell+behaviour%3A+I.+Simple+step+cues%5BTitle%5D)+AND+%22Development%22%5BJournal%5D)">Topographical control of cell behaviour: I. Simple step cues.</a> Development. 99 (3), 439-448 (1987).</li>
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<li>Corbin, E. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Tunable+and+reversible+substrate+stiffness+reveals+a+dynamic+mechanosensitivity+of+cardiomyocytes%5BTitle%5D)+AND+%22ACS+Appl+Mater+Interfaces%22%5BJournal%5D)">Tunable and reversible substrate stiffness reveals a dynamic mechanosensitivity of cardiomyocytes.</a> ACS Appl Mater Interfaces. 11 (23), 20603-20614 (2019).</li>
<li>Bouhrira, N., Vite, A., Margulies, K. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Distinct+cytoskeletal+regulators+of+mechanical+memory+in+cardiac+fibroblasts+and+cardiomyocytes%5BTitle%5D)+AND+%22Basic+Res+Cardiol%22%5BJournal%5D)">Distinct cytoskeletal regulators of mechanical memory in cardiac fibroblasts and cardiomyocytes.</a> Basic Res Cardiol. 119 (2), 277-289 (2024).</li>
<li>Lee, B. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adult+human+cardiomyocyte+mechanics+in+osteogenesis+imperfecta%5BTitle%5D)+AND+%22Am+J+Physiol+Heart+Circ+Physiol%22%5BJournal%5D)">Adult human cardiomyocyte mechanics in osteogenesis imperfecta.</a> Am J Physiol Heart Circ Physiol. 325 (4), H814-H821 (2023).</li>
<li>Cao, Z., Ball, J. K., Lateef, A. H., Virgile, C. P., Corbin, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Biomimetic+substrate+to+probe+dynamic+interplay+of+topography+and+stiffness+on+cardiac+fibroblast+activation%5BTitle%5D)+AND+%22ACS+Omega%22%5BJournal%5D)">Biomimetic substrate to probe dynamic interplay of topography and stiffness on cardiac fibroblast activation.</a> ACS Omega. 8 (6), 5406-5414 (2023).</li>
<li>Clark, A. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Magnetic+field+tuning+of+mechanical+properties+of+ultrasoft+PDMSbased+magnetorheological+elastomers+for+biological+applications%5BTitle%5D)+AND+%22Multifunct+Mater%22%5BJournal%5D)">Magnetic field tuning of mechanical properties of ultrasoft PDMSbased magnetorheological elastomers for biological applications.</a> Multifunct Mater. 4 (3), 035001(2021).</li>
<li>Kim, Y., Kwon, C., Jeon, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Genetically+engineered+phage+induced+selective+H9c2+cardiomyocytes+patterning+in+PDMS+microgrooves%5BTitle%5D)+AND+%22Materials%22%5BJournal%5D)">Genetically engineered phage induced selective H9c2 cardiomyocytes patterning in PDMS microgrooves.</a> Materials. 10 (8), 973(2017).</li>
<li>Fujiwara, Y., Deguchi, K., Miki, K., Nishimoto, T., Yoshida, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+method+for+contraction+force+measurement+of+hiPSCderived+engineered+cardiac+tissues%5BTitle%5D)+AND+%22Methods+Mol+Biol%22%5BJournal%5D)">A method for contraction force measurement of hiPSCderived engineered cardiac tissues.</a> Methods Mol Biol. 2320, 171-180 (2021).</li>
<li>Dirar, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Activation+and+degranulation+of+CART+cells+using+engineered+antigenpresenting+cell+surfaces%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Activation and degranulation of CART cells using engineered antigenpresenting cell surfaces.</a> PLoS One. 15 (9), e0238819(2020).</li>
<li>Kim, C., Kim, H., Park, H., Lee, K. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Controlling+the+porous+structure+of+alginate+ferrogel+for+anticancer+drug+delivery+under+magnetic+stimulation%5BTitle%5D)+AND+%22Carbohydr+Polym%22%5BJournal%5D)">Controlling the porous structure of alginate ferrogel for anticancer drug delivery under magnetic stimulation.</a> Carbohydr Polym. 223, 115045(2019).</li>
<li>Fan, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Recent+advances+of+magnetic+nanomaterials+in+bone+tissue+repair%5BTitle%5D)+AND+%22Front+Chem%22%5BJournal%5D)">Recent advances of magnetic nanomaterials in bone tissue repair.</a> Front Chem. 8, 745(2020).</li>
<li>GonzalezRico, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Tuning+the+cell+and+biological+tissue+environment+through+magnetoactive+materials%5BTitle%5D)+AND+%22Appl+Sci%22%5BJournal%5D)">Tuning the cell and biological tissue environment through magnetoactive materials.</a> Appl Sci. 11 (18), 8746(2021).</li>
<li>AntmanPassig, M., Shefi, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Remote+magnetic+orientation+of+3D+collagen+hydrogels+for+directed+neuronal+regeneration%5BTitle%5D)+AND+%22Nano+Lett%22%5BJournal%5D)">Remote magnetic orientation of 3D collagen hydrogels for directed neuronal regeneration.</a> Nano Lett. 16 (4), 2567-2573 (2016).</li>
</ol>

<table><tbody><tr><td>555 Alexa Fluor anti-camundongo anticorpo secundário de camundongos &nbsp;</td><td>ThermoFisher Scientific</td><td>A-21422</td><td>https://www.thermofisher.com/antibody/product/Goat-anti-Mouse-IgG-H-L-Cross-Adsorbed-Secondary-Antibody-Polyclonal/A-21422</td></tr><tr><td>Lente 20x mergulhada</td><td>Zeiss</td><td>421452-9800-000</td><td>https://www.micro-shop.zeiss.com/zh/us/shop/objectives/421452-9800-000/Objective-W-Plan-Apochromat-20x-1.0-DIC-M27-75mm</td></tr><tr><td>Antenas de 35 mm</td><td>ThermoFisher Scientific</td><td>130180</td><td>https://www.thermofisher.com/search/results?query=130180&focusarea=Search%20All</td></tr><tr><td>Anticorpo primário anti-alfa actinina</td><td>Abcam</td><td>EP2528Y</td><td>https://www.abcam.com/en-us/products/primary-antibodies/alpha-actinin-actn1-antibody-ep2528y-ab81265?srsltid=AfmBOoq4HwHixqCheQwwq VVbjJ3ogKUmZxa4GtG 3SyllFzWvb5XPnwgB</td></tr><tr><td>Microsferas em pó de ferro carbonilo</td><td>chemicalstore.com</td><td>CIPMS</td><td>https://ironpowders.com/types-of-iron-powder/cipms-carbonyl-iron-powder/</td></tr><tr><td>Coloração DAPI</td><td>ThermoFisher Scientific</td><td>D1306</td><td>https://www.thermofisher.com/order/catalog/product/D1306</td></tr><tr><td>Dessecador</td><td>Fisher Scientific</td><td>08-642-7</td><td>https://www.fishersci.com/shop/products/nalgene-transparent-polycarbonate-classic-design-desiccator/086427#?keyword=</td></tr><tr><td>Grade de difração</td><td>Thor Labs</td><td>GR2550-45031</td><td>https://www.thorlabs.com/thorproduct.cfm?partnumber=GR2550-45031</td></tr><tr><td>Gel Ecoflex parte A & B</td><td>Smooth On</td><td>Ecoflex&nbsp; GEL</td><td>https://www.smooth-on.com/products/ecoflex-gel/</td></tr><tr><td>Limpador de plasma expandido para mesa</td><td>Sigma Aldrich</td><td>Z561657-1EA</td><td>https://www.sigmaaldrich.com/US/en/product/aldrich/z561657</td></tr><tr><td>Fibronectina do plasma bovino</td><td>Sigma Aldrich</td><td>F1141</td><td>https://www.sigmaaldrich.com/US/en/product/sigma/f1141</td></tr><tr><td>Flacktek DAC150 SpeedMixer&nbsp;</td><td>Flacktek</td><td>DAC150</td><td>https://flacktek.com/products/small-flacktek/</td></tr><tr><td>ImageJ Software</td><td>NIH</td><td></td><td>https://imagej.net/ij/download.html</td></tr><tr><td>Álcool isopropílico</td><td>Amazon</td><td></td><td>https://www.amazon.com/Isopropyl-Alcohol-Grade-Anhydrous-gallon/dp/B01M6YK5I4/ref=sr_1_1_sspa?dib=eyJ2IjoiMSJ9.xiBzaAiE5lSJqXz ZhS_JhDh8zEHgkMzrbsyosvyBpS Jjufk90Gt79vk1jOZxq_mG3GKeN8 KT1ulag3ZuMShbWoEOCQUy6txx uGz4709PyUY5YWyeR2GS0PBNx dbmEdWvgmCYiDBLf8E2prhImZW l98MoWlVDU4lg1BtcHTTSOwdg87 5l6Ia3YLPs_0alILBXFsizwwnJodR 8D60ZggK5AHBfCfPadXgfJ-9hUE7 uIto.V3Vf62Lnprtq2u5PRO SbUFHnPOj1FvD7np7uBge6NuU& dib_tag=se& hvadid=598724400239 & hvdev=c& hvlocphy=9007460& HVn ETW=G& hvqmt=e & ; hvrand=1609157 209668794739& hvtargid=kwd-3203 62094308& hydadcr=5273_13 227703& palavras-chave=isopropilo+álcool+ amazon& MCID=9F9D3252906531 3EA89AB7766D9C244C& qid=1740 675756& sr=8-1-spons&; sp_csd=d2l kZ2V0TmFtZT1zcF9hdGY& psc=1</td></tr><tr><td>Ímã</td><td>Magnetismo CMS</td><td>ND05578-45N</td><td>https://www.magnet4sale.com/n45-neodymium-disc-magnet-1-1-2x1-4-35-lb-pull-strong-magnet/</td></tr><tr><td>Microscópio vertical Nikon Eclipse 80i</td><td>Nikon</td><td>Eclipse 80i</td><td></td></tr><tr><td>Forno</td><td>Fisher Scientific</td><td>15-103-0503</td><td>https://www.fishersci.com/shop/products/fisher-scientific-isotemp-general-purpose-heating-drying-ovens/p-4935382</td></tr><tr><td>Diluente de silicone</td><td>Smooth On</td><td>Diluente de silicone</td><td>https://www.smooth-on.com/products/silicone-thinner/</td></tr><tr><td>Mancha de Actina SiR</td><td>Citoesqueleto</td><td>CY-SC001</td><td>https://www.cytoskeleton.com/sir-actin</td></tr><tr><td>Sylgard 184 PDMS (Base + agente de cura)</td><td>Adesivos Ellsworth</td><td>2065622</td><td>https://www.ellsworth.com/products/by-market/consumer-products/encapsulants/silicone/dow-sylgard-184-silicone-encapsulant-clear-3.9-kg-kit/</td></tr><tr><td>Sylgard 527 PDMS parte A & B</td><td>Adesivos Ellsworth</td><td>1696742</td><td>https://www.ellsworth.com/products/by-market/consumer-products/encapsulants/silicone/dow-sylgard-527-silicone-dielectric-gel-clear-0.9-kg-kit/</td></tr><tr><td>Pipetas de transferência</td><td>Fisher Scientific</td><td>13-711-5AMMD</td><td>https://www.fishersci.com/shop/products/transfer-pipette-32/137115AMMD?searchHijack=true&searchTerm=13 -711-5AMMD& searchType= RAPID& matchedCatNo=13 -711-5AMMD</td></tr><tr><td>Tricloro -(1H, 1H, 2H, 2H-perfluorooctil) silano&nbsp;</td><td>Sigma Aldrich</td><td>448931</td><td>https://www.sigmaaldrich.com/US/en/product/aldrich/448931</td></tr></tbody></table>

elastômeros magneto-reológicos micropadronizados para impulsionar mudanças no alinhamento de cardiomiócitos

Micropatterned Magneto-Rheological Elastomers to Drive Changes in Cardiomyocyte Alignment

Elastômeros magneto-reológicos micropadronizados para impulsionar mudanças no alinhamento de cardiomiócitos

Substrate-associated cues, such as mechanical and topographic, profoundly influence cellular response. However, much of the foundational research employs ...

micropatterned-magneto-rheological-elastomers-to-drive-changes

Research

JoVE Journal

Bioengineering

Bioengenharia

Neste protocolo, demonstramos a fabricação de substratos biomiméticos de cultura de células cardíacas feitos de dois materiais poliméricos distintos usando litografia de força capilar.  Os métodos descritos fornecem uma técnica escalável e econômica para projetar a estrutura e a função dos tecidos cardíacos macroscópicos para aplicações in vitro e in vivo.

Capilar Força Litografia de Engenharia tecido cardíaco

A geração de alinhados tecido do miocárdio é um requisito fundamental para a adaptação dos avanços recentes em biologia de células-tronco para fins clinicamente úteis. Descrevemos aqui uma abordagem impressão microcontact para o controlo preciso da forma e função celular. Usando populações altamente purificadas de células-tronco embrionárias derivadas progenitores cardíacos, nós, então, gerar tecido funcional anisotrópica do miocárdio.

Geração de Alinhados tecido funcional do miocárdio através de impressão microcontact

Apresentamos um novo hidrogel de poliacrilamida, chamada hidroxi-PAAm, que permite uma ligação direta das proteínas da MEC com custo ou experiência mínima. A combinação de hidroxi-PAAm hidrogéis com impressão microcontact facilita o controlo independente das muitas pistas de o microambiente celular natural para estudar mechanostransduction celular.

Preparação de Hidroxi-PAAm hidrogéis para dissociar os efeitos de Mecanotransdução Sugestões

Este protocolo descreve um método novo para criar um modelo de elementos finitos espacialmente detalhada da arquitetura intracelular do cardiomyocytes de imagens de microscopia confocal e microscopia eletrônica. O poder deste modelo espacialmente detalhada é demonstrado usando estudos de caso em sinalização de cálcio e Bioenergética.

Criando um modelo geométrico de elementos finitos estruturalmente realista de um casos para estudar o papel da arquitetura celular em casos de biologia de sistemas

Aqui nós apresentamos um protocolo para a formação de uma população celular usando estímulos elétricos e mecânicos, emulando a fisiologia cardíaca. Esta estimulação eletromecânica aumenta o potencial de cardiomyogenic das células tratadas e é uma estratégia promissora para mais terapia celular, modelagem de doença e despistagem de drogas.

Estimulação simultânea de elétrica e mecânica para aumentar o potencial de Cardiomyogenic das células

Um andaime bioinspirado é fabricado por uma técnica de fotolitografia macia usando hidrogéis mecanicamente robustos e eletricamente condutores. Os hidrogéis micropadronizados fornecem alinhamento direcional das células cardiomiócitos, resultando em uma direção personalizada de atuação. Microeletrodos flexíveis também são integrados ao andaime para trazer controle elétrico para um tecido cardíaco auto-atuante.

Robô macio bioinspirado com microeletrodos incorporados

Este protocolo fornece métodos detalhados descrevendo a fabricação e a implementação de uma plataforma de ajuste pós-carga baseada em magnéticos para tecidos cardíacos projetados.

Ajuste magnético da pós-carga em tecidos cardíacos projetados

Este protocolo introduz um método de micropadronização livre de litografia que é simples e acessível para aqueles com um fundo de bioengenharia limitado. Este método utiliza estêncil personalizado de corte a laser para micropadrão proteínas de matriz extracelular em uma forma de interesse para modulação de morfologias celulares. O procedimento de micropadronização é demonstrado usando cardiomiócitos derivados de células-tronco pluripotentes induzidas.

Micropadronização simples de células únicas sem litografia usando estêncil cortado a laser

O presente protocolo gera tecidos cardíacos projetados em forma de malha contendo células cardiovasculares derivadas de células-tronco pluripotentes induzidas pelo homem para permitir a investigação da terapia de implantação celular para doenças cardíacas.

Preparação de tecidos cardíacos projetados em forma de malha derivados de células iPS humanas para reparo de miocárdio in vivo

Este protocolo visa descrever uma nova metodologia para medir a taxa de disparo cardíaco intrínseco usando o registro de matriz de microeletrínda de todo o tecido do nódulo sinoatrial para identificar defeitos de marcação de ritmo em camundongos. Agentes farmacológicos também podem ser introduzidos neste método para estudar seus efeitos no ritmo intrínseco.

Microelectrode Array Gravação da Taxa de Disparo de Nó Sinoatrial para identificar defeitos intrínsecos de marcação cardíaca em camundongos

Este trabalho apresenta um protocolo flexível para utilizar a tecnologia de superfícies contraíveis elastoméricas rotuladas fluorescentes (FLECS) em formato de microwell para quantificação simplificada e prática de forças contraídas unicelulares com base em deslocamentos visualizados de micropatterns de proteína fluorescente.

Análise simplificada e de alto rendimento da contractilidade unicelular usando Elastômeros Micropatterned

Aprenda a fabricar elastômeros magneto-reológicos micropadronizados para estudar o alinhamento de cardiomiócitos em resposta a mudanças dinâmicas de rigidez. Esta plataforma auxilia pesquisadores em cardiologia.

Elastômeros magneto-reológicos micropadronizados para impulsionar mudanças no alinhamento de cardiomiócitos

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