Electrical coupling of isolated cardiomyocyte clusters grown on aligned conductive nanofibrous meshes for their synchronized beating.

Myocardial infarction is often associated with abnormalities in electrical function due to a massive loss of functioning cardiomyocytes. This work develops a mesh, consisting of aligned composite nanofibers of polyaniline (PANI) and poly(lactic-co-glycolic acid) (PLGA), as an electrically active scaffold for coordinating the beatings of the cultured cardiomyocytes synchronously. Following doping by HCl, the electrospun fibers could be transformed into a conductive form carrying positive charges, which could then attract negatively charged adhesive proteins (i.e. fibronectin and laminin) and enhance cell adhesion. During incubation, the adhered cardiomyocytes became associated with each other and formed isolated cell clusters; the cells within each cluster elongated and aligned their morphology along the major axis of the fibrous mesh. After culture, expression of the gap-junction protein connexin 43 was clearly observed intercellularly in isolated clusters. All of the cardiomyocytes within each cluster beat synchronously, implying that the coupling between the cells was fully developed. Additionally, the beating rates among these isolated cell clusters could be synchronized via an electrical stimulation designed to imitate that generated in a native heart. Importantly, improving the impaired heart function depends on electrical coupling between the engrafted cells and the host myocardium to ensure their synchronized beating.

[1]  Xuesi Chen,et al.  Electroactive oligoaniline-containing self-assembled monolayers for tissue engineering applications. , 2007, Biomacromolecules.

[2]  E. Lakatta,et al.  Excitation‐contraction coupling in the heart: the state of the question 1 , 1992, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[3]  P. G. Rasmussen,et al.  Characterization of solution and solid state properties of undoped and doped polyanilines processed from hexafluoro-2-propanol , 1996 .

[4]  C. Murry,et al.  Electromechanical coupling between skeletal and cardiac muscle. Implications for infarct repair. , 2000 .

[5]  Zhao Ping In situ FTIR–attenuated total reflection spectroscopic investigations on the base–acid transitions of polyaniline. Base–acid transition in the emeraldine form of polyaniline , 1996 .

[6]  R. Weisel,et al.  Survival and function of bioengineered cardiac grafts. , 1999, Circulation.

[7]  H. Sung,et al.  The characteristics, cellular uptake and intracellular trafficking of nanoparticles made of hydrophobically-modified chitosan. , 2010, Journal of controlled release : official journal of the Controlled Release Society.

[8]  Alan G. MacDiarmid,et al.  ‘Polyaniline’: Protonic acid doping of the emeraldine form to the metallic regime , 1986 .

[9]  C. Chuang,et al.  Magnetic-nanoparticle-modified paclitaxel for targeted therapy for prostate cancer. , 2010, Biomaterials.

[10]  Andreas Hess,et al.  Engineered heart tissue grafts improve systolic and diastolic function in infarcted rat hearts , 2006, Nature Medicine.

[11]  A R Boccaccini,et al.  Myocardial tissue engineering: a review , 2007, Journal of tissue engineering and regenerative medicine.

[12]  E. Erdem,et al.  Synthesis and characterization of malonic acid-doped polyaniline , 1997 .

[13]  Yen Wei,et al.  Electrospinning polyaniline-contained gelatin nanofibers for tissue engineering applications. , 2006, Biomaterials.

[14]  E. Entcheva,et al.  Electrospun fine-textured scaffolds for heart tissue constructs. , 2005, Biomaterials.

[15]  A. Heeger,et al.  Poly-p-phenyleneamineimine: synthesis and comparison to polyaniline , 1987 .

[16]  Milica Radisic,et al.  Challenges in cardiac tissue engineering. , 2010, Tissue engineering. Part B, Reviews.

[17]  D E H Tee,et al.  Book Review: Culture of Animal Cells: A Manual of Basic Technique , 1984 .

[18]  K. Neoh,et al.  Polyaniline with high intrinsic oxidation state , 1993 .

[19]  L. Reinlib,et al.  Cell transplantation as future therapy for cardiovascular disease?: A workshop of the National Heart, Lung, and Blood Institute. , 2000, Circulation.

[20]  J. Vacanti,et al.  Contractile cardiac grafts using a novel nanofibrous mesh. , 2004, Biomaterials.

[21]  J. Vacanti,et al.  A biodegradable nanofiber scaffold by electrospinning and its potential for bone tissue engineering. , 2003, Biomaterials.

[22]  K. Kern,et al.  Electrospinning of Diphenylalanine Nanotubes , 2008 .

[23]  Chun-Wen Hsiao,et al.  Magnetically directed self-assembly of electrospun superparamagnetic fibrous bundles to form three-dimensional tissues with a highly ordered architecture. , 2011, Tissue engineering. Part C, Methods.

[24]  C. Montero-Menei,et al.  Combining adult stem cells and polymeric devices for tissue engineering in infarcted myocardium. , 2012, Biomaterials.

[25]  Koichi Kawahara,et al.  Changes in the fluctuation of interbeat intervals in spontaneously beating cultured cardiac myocytes: experimental and modeling studies , 2002, Biological Cybernetics.

[26]  John R. Reynolds,et al.  Multiply Colored Electrochromic Carbazole-Based Polymers , 1997 .

[27]  Jae Young Lee,et al.  Polypyrrole-coated electrospun PLGA nanofibers for neural tissue applications. , 2009, Biomaterials.

[28]  Heungsoo Shin,et al.  The stimulation of myoblast differentiation by electrically conductive sub-micron fibers. , 2009, Biomaterials.

[29]  M. Heath,et al.  Ion-specific effects on the interaction between fibronectin and negatively charged mica surfaces. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[30]  L. Niklason,et al.  Scaffold-free vascular tissue engineering using bioprinting. , 2009, Biomaterials.

[31]  Chien-Chung Han,et al.  Combination of Electrochemistry with Concurrent Reduction and Substitution Chemistry To Provide a Facile and Versatile Tool for Preparing Highly Functionalized Polyanilines , 1999 .

[32]  D. Atsma,et al.  Forced Alignment of Mesenchymal Stem Cells Undergoing Cardiomyogenic Differentiation Affects Functional Integration With Cardiomyocyte Cultures , 2008, Circulation research.

[33]  B D Boyan,et al.  Role of material surfaces in regulating bone and cartilage cell response. , 1996, Biomaterials.

[34]  Holly M. Brown-Borg,et al.  Culture of Animal Cells: A Manual of Basic Technique, R. Ian Freshney (Ed.). Wiley-Liss, Inc., New York (1994) , 1995 .

[35]  Fei Yang,et al.  The impact of PLGA scaffold orientation on in vitro cartilage regeneration. , 2012, Biomaterials.

[36]  Guoping Chen,et al.  The influence of structural design of PLGA/collagen hybrid scaffolds in cartilage tissue engineering. , 2010, Biomaterials.

[37]  C. Murry,et al.  Electromechanical Coupling between Skeletal and Cardiac Muscle , 2000, The Journal of cell biology.

[38]  E. Ehler,et al.  Dilated cardiomyopathy: a disease of the intercalated disc? , 2003, Trends in cardiovascular medicine.

[39]  Younan Xia,et al.  Understanding the role of surface charges in cellular adsorption versus internalization by selectively removing gold nanoparticles on the cell surface with a I2/KI etchant. , 2009, Nano letters.

[40]  Milica Radisic,et al.  Functional assembly of engineered myocardium by electrical stimulation of cardiac myocytes cultured on scaffolds , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[41]  Sook Hee Ku,et al.  Synergic effects of nanofiber alignment and electroactivity on myoblast differentiation. , 2012, Biomaterials.

[42]  Seeram Ramakrishna,et al.  Electrospun conducting polymer nanofibers and electrical stimulation of nerve stem cells. , 2011, Journal of bioscience and bioengineering.

[43]  Younan Xia,et al.  Injectable PLGA porous beads cellularized by hAFSCs for cellular cardiomyoplasty. , 2012, Biomaterials.

[44]  Kimberly A Woodhouse,et al.  Culture on electrospun polyurethane scaffolds decreases atrial natriuretic peptide expression by cardiomyocytes in vitro. , 2008, Biomaterials.

[45]  A. Epstein,et al.  'Synthetic metals': a novel role for organic polymers , 1989, Images of the Twenty-First Century. Proceedings of the Annual International Engineering in Medicine and Biology Society,.

[46]  Jeroen J. Bax,et al.  Impact of Viability, Ischemia, Scar Tissue, and Revascularization on Outcome After Aborted Sudden Death , 2003, Circulation.

[47]  Yen Wei,et al.  Polyaniline, an electroactive polymer, supports adhesion and proliferation of cardiac myoblasts , 2006, Journal of biomaterials science. Polymer edition.

[48]  M. Textor,et al.  Effects of ionic strength and surface charge on protein adsorption at PEGylated surfaces. , 2005, The journal of physical chemistry. B.

[49]  S. Vaidya Facile methods for enhancement of conductivity in polyaniline , 2009 .

[50]  H. Krug,et al.  Oops they did it again! Carbon nanotubes hoax scientists in viability assays. , 2006, Nano letters.

[51]  A. MacDiarmid,et al.  "Synthetic Metals": A Novel Role for Organic Polymers (Nobel Lecture). , 2001, Angewandte Chemie.

[52]  Jin-Oh You,et al.  Nanoengineering the heart: conductive scaffolds enhance connexin 43 expression. , 2011, Nano letters.

[53]  H. Haider,et al.  Sca-1+ Stem Cell Survival and Engraftment in the Infarcted Heart: Dual Role for Preconditioning-Induced Connexin-43 , 2009, Circulation.

[54]  Ajay M Shah,et al.  Regulation of cardiac contractile function by troponin I phosphorylation. , 2005, Cardiovascular research.

[55]  Izumi Ichinose,et al.  Biocompatibility of layer-by-layer self-assembled nanofilm on silicone rubber for neurons , 2003, Journal of Neuroscience Methods.

[56]  Milica Radisic,et al.  Interactive effects of surface topography and pulsatile electrical field stimulation on orientation and elongation of fibroblasts and cardiomyocytes. , 2007, Biomaterials.

[57]  J. A. Hubbell,et al.  Poly(ethylene oxide)-graft-poly(L-lysine) copolymers to enhance the biocompatibility of poly(L-lysine)-alginate microcapsule membranes. , 1992, Biomaterials.