Strategies and Processes to Decellularize and Recellularize Hearts to Generate Functional Organs and Reduce the Risk of Thrombosis

Heart failure is one of the leading causes of death in the United States. Current therapies, such as heart transplants and bioartificial hearts, are helpful, but not optimal. Decellularization of porcine whole hearts followed by recellularization with patient-specific human cells may provide the ultimate solution for patients with heart failure. Great progress has been made in the development of efficient processes for decellularization, and the design of automated bioreactors. Challenges remain in selecting and culturing cells, growing the cells on the decellularized scaffolds without contamination, characterizing the regenerated organs, and preventing thrombosis. Various strategies have been proposed to prevent thrombosis of blood-contacting devices, including reendothelization and the creation of nonfouling surfaces using surface modification technologies. This review discusses the progress and remaining challenges involved with recellularizing whole hearts, focusing on the prevention of thrombosis.

[1]  Gino Gerosa,et al.  Cell characterization of porcine aortic valve and decellularized leaflets repopulated with aortic valve interstitial cells: the VESALIO Project (Vitalitate Exornatum Succedaneum Aorticum Labore Ingenioso Obtenibitur). , 2003, The Annals of thoracic surgery.

[2]  T. Jensen,et al.  A rapid lung de-cellularization protocol supports embryonic stem cell differentiation in vitro and following implantation. , 2012, Tissue engineering. Part C, Methods.

[3]  J. Hubbell,et al.  An RGD spacing of 440 nm is sufficient for integrin alpha V beta 3- mediated fibroblast spreading and 140 nm for focal contact and stress fiber formation , 1991, The Journal of cell biology.

[4]  Aline M. Betancourt,et al.  A nonhuman primate model of lung regeneration: detergent-mediated decellularization and initial in vitro recellularization with mesenchymal stem cells. , 2012, Tissue engineering. Part A.

[5]  R Langer,et al.  Characterization and development of RGD-peptide-modified poly(lactic acid-co-lysine) as an interactive, resorbable biomaterial. , 1997, Journal of biomedical materials research.

[6]  K. Hochedlinger,et al.  Harnessing the potential of induced pluripotent stem cells for regenerative medicine , 2011, Nature Cell Biology.

[7]  E. Sparrow,et al.  Bioreactor for the reconstitution of a decellularized vascular matrix of biological origin , 2011 .

[8]  D. Fawcett,et al.  Engineering a Biocompatible Scaffold with Either Micrometre or Nanometre Scale Surface Topography for Promoting Protein Adsorption and Cellular Response , 2013, International journal of biomaterials.

[9]  Lei Yang,et al.  Hear the beat: decellularized mouse heart regenerated with human induced pluripotent stem cells , 2014, Expert review of cardiovascular therapy.

[10]  P. Nguyen,et al.  Safe Genetic Modification of Cardiac Stem Cells Using a Site-Specific Integration Technique , 2012, Circulation.

[11]  Ronald M. McLaughlin,et al.  Myocardial scaffold-based cardiac tissue engineering: application of coordinated mechanical and electrical stimulations. , 2013, Langmuir : the ACS journal of surfaces and colloids.

[12]  A. Moorman,et al.  Role of bone morphogenetic proteins in cardiac differentiation. , 2007, Cardiovascular research.

[13]  Buddy D Ratner,et al.  The catastrophe revisited: blood compatibility in the 21st Century. , 2007, Biomaterials.

[14]  Sandra R. Smith,et al.  Growth factor effects on cells of the vascular wall: a survey. , 1993, Growth factors.

[15]  B. Conklin,et al.  Development and evaluation of a novel decellularized vascular xenograft. , 2002, Medical engineering & physics.

[16]  G. Lemon,et al.  Verification of cell viability in bioengineered tissues and organs before clinical transplantation. , 2013, Biomaterials.

[17]  Robert Langer,et al.  Synthesis and RGD peptide modification of a new biodegradable copolymer: poly(lactic acid-co-lysine) , 1993 .

[18]  Stuart L. Cooper,et al.  Synthesis, surface, and cell-adhesion properties of polyurethanes containing covalently grafted RGD-peptides. , 1994 .

[19]  L. Popescu,et al.  Heterocellular communication in the heart: electron tomography of telocyte–myocyte junctions , 2011, Journal of cellular and molecular medicine.

[20]  L. Niklason,et al.  Novel utilization of serum in tissue decellularization. , 2010, Tissue engineering. Part C, Methods.

[21]  T. Ashikaga,et al.  The effects of storage and sterilization on de-cellularized and re-cellularized whole lung. , 2013, Biomaterials.

[22]  Stephen F Badylak,et al.  Quantification of DNA in biologic scaffold materials. , 2009, The Journal of surgical research.

[23]  A. Lichtenberg,et al.  The quest for an optimized protocol for whole-heart decellularization: a comparison of three popular and a novel decellularization technique and their diverse effects on crucial extracellular matrix qualities. , 2011, Tissue engineering. Part C, Methods.

[24]  A. Perets,et al.  Enhancing the vascularization of three-dimensional porous alginate scaffolds by incorporating controlled release basic fibroblast growth factor microspheres. , 2003, Journal of biomedical materials research. Part A.

[25]  David J. Mooney,et al.  Controlled growth factor release from synthetic extracellular matrices , 2000, Nature.

[26]  A. Simmons,et al.  The modulation of platelet and endothelial cell adhesion to vascular graft materials by perlecan. , 2009, Biomaterials.

[27]  Application of a rotating bioreactor consisting of low-cost and ready-to-use medical disposables for in vitro evaluation of the endothelialization efficiency of small-caliber vascular prostheses. , 2013, Journal of biomedical materials research. Part B, Applied biomaterials.

[28]  S. Hanson,et al.  Blood flow and antithrombotic drug effects. , 1998, American heart journal.

[29]  Anthony Atala,et al.  Decellularization methods of porcine kidneys for whole organ engineering using a high-throughput system. , 2012, Biomaterials.

[30]  J. Copeland,et al.  Standardized methods to quantify thrombogenicity of blood-contacting materials via thromboelastography. , 2012, Journal of biomedical materials research. Part B, Applied biomaterials.

[31]  M. Hiles,et al.  Virus safety of a porcine‐derived medical device: Evaluation of a viral inactivation method , 2002, Biotechnology and bioengineering.

[32]  M. Weiss,et al.  Stem cells in the umbilical cord , 2006, Stem Cell Reviews.

[33]  R. Schnaar,et al.  Covalent attachment of an Arg-Gly-Asp sequence peptide to derivatizable polyacrylamide surfaces: support of fibroblast adhesion and long-term growth. , 1988, Analytical biochemistry.

[34]  Harald C Ott,et al.  Perspectives on whole-organ assembly: moving toward transplantation on demand. , 2012, The Journal of clinical investigation.

[35]  J. Leor,et al.  Modulation of cardiac macrophages by phosphatidylserine-presenting liposomes improves infarct repair , 2011, Proceedings of the National Academy of Sciences.

[36]  Elizabeth A. Calle,et al.  Procedure for lung engineering. , 2011, Journal of visualized experiments : JoVE.

[37]  Zhen W. Zhuang,et al.  Tissue-Engineered Lungs for in Vivo Implantation , 2010, Science.

[38]  D. Zopf,et al.  Effects of sterilization on an extracellular matrix scaffold: Part I. Composition and matrix architecture , 2007, Journal of materials science. Materials in medicine.

[39]  Mark Turmaine,et al.  Discarded human kidneys as a source of ECM scaffold for kidney regeneration technologies. , 2013, Biomaterials.

[40]  S. Schwartz,et al.  Embryonic stem cell trials for macular degeneration: a preliminary report , 2012, The Lancet.

[41]  Stephen F Badylak,et al.  Decellularization of tissues and organs. , 2006, Biomaterials.

[42]  David Milan,et al.  Inefficient Reprogramming of Fibroblasts into Cardiomyocytes Using Gata4, Mef2c, and Tbx5 , 2012, Circulation research.

[43]  N. Frangogiannis,et al.  Fibroblasts in post-infarction inflammation and cardiac repair. , 2013, Biochimica et biophysica acta.

[44]  Korkut Uygun,et al.  Whole-organ tissue engineering: decellularization and recellularization of three-dimensional matrix scaffolds. , 2011, Annual review of biomedical engineering.

[45]  N. Frangogiannis,et al.  Transforming growth factor (TGF)-β signaling in cardiac remodeling. , 2011, Journal of molecular and cellular cardiology.

[46]  Kerry A. Daly,et al.  Biologic scaffolds for constructive tissue remodeling. , 2011, Biomaterials.

[47]  Patrick S. Chang,et al.  Spatial organization and mechanical properties of the pericellular matrix on chondrocytes. , 2013, Biophysical journal.

[48]  Adam Byron,et al.  Defining the extracellular matrix using proteomics , 2013, International journal of experimental pathology.

[49]  Anthony Callanan,et al.  Comparison of methods for whole-organ decellularization in tissue engineering of bioartificial organs. , 2013, Tissue engineering. Part B, Reviews.

[50]  Martin Ehrbar,et al.  Cell‐demanded release of VEGF from synthetic, biointeractive cell‐ingrowth matrices for vascularized tissue growth , 2003, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[51]  Robert Langer,et al.  Vascular Catheters with a Nonleaching Poly-Sulfobetaine Surface Modification Reduce Thrombus Formation and Microbial Attachment , 2012, Science Translational Medicine.

[52]  Alison P McGuigan,et al.  The thrombogenicity of human umbilical vein endothelial cell seeded collagen modules. , 2008, Biomaterials.

[53]  Stephen F Badylak,et al.  RETRACTED: Engineered whole organs and complex tissues , 2012, The Lancet.

[54]  H. Wendel,et al.  A novel in vitro model for preclinical testing of the hemocompatibility of intravascular stents according to ISO 10993-4 , 2011, Journal of materials science. Materials in medicine.

[55]  J. Hubbell,et al.  Covalent surface immobilization of Arg-Gly-Asp- and Tyr-Ile-Gly-Ser-Arg-containing peptides to obtain well-defined cell-adhesive substrates. , 1990, Analytical biochemistry.

[56]  Lei Yang,et al.  Repopulation of decellularized mouse heart with human induced pluripotent stem cell-derived cardiovascular progenitor cells , 2013, Nature Communications.

[57]  Stephen F Badylak,et al.  Immune response to biologic scaffold materials. , 2008, Seminars in Immunology.

[58]  S. Mohammad,et al.  An In-vitro Model to Study Device-induced Thrombosis and Embolism: , 2000, Thrombosis and Haemostasis.

[59]  Xi Ren,et al.  Perfusion decellularization of whole organs , 2014, Nature Protocols.

[60]  C. Harley,et al.  Telomeres shorten during ageing of human fibroblasts , 1990, Nature.

[61]  N. Frangogiannis,et al.  The role of TGF-β Signaling in Myocardial Infarction and Cardiac Remodeling , 2007 .

[62]  C. Lengner iPS cell technology in regenerative medicine , 2010, Annals of the New York Academy of Sciences.

[63]  V. Vedantham,et al.  Direct Reprogramming of Fibroblasts into Functional Cardiomyocytes by Defined Factors , 2010, Cell.

[64]  S. Sukavaneshvar Assessment and Management of Vascular Implant Thrombogenecity , 2008 .

[65]  Hug Aubin,et al.  In vivo functional performance and structural maturation of decellularised allogenic aortic valves in the subcoronary position. , 2010, European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery.

[66]  C. Howe,et al.  Removal of unbound sodium dodecyl sulfate (SDS) from proteins in solution by electrophoresis through triton x-100-agarose. , 1978, Journal of immunological methods.

[67]  Artur Lichtenberg,et al.  Preclinical Testing of Tissue-Engineered Heart Valves Re-Endothelialized Under Simulated Physiological Conditions , 2006, Circulation.

[68]  E. Wolner,et al.  Decellularization does not eliminate thrombogenicity and inflammatory stimulation in tissue-engineered porcine heart valves. , 2006, The Journal of heart valve disease.

[69]  Claire Robertson,et al.  Optical imaging predicts mechanical properties during decellularization of cardiac tissue. , 2013, Tissue engineering. Part C, Methods.

[70]  Matthew J. Robertson,et al.  Optimizing Recellularization of Whole Decellularized Heart Extracellular Matrix , 2014, PloS one.

[71]  H. Augustin,et al.  Angiopoietins: a link between angiogenesis and inflammation. , 2006, Trends in immunology.

[72]  R. C. Thomson,et al.  Improvements in GORE-TEX 1 Vascular Graft Performance by Carmeda 1 BioActive Surface Heparin Immobilization , 2012 .

[73]  Johanna Andrae,et al.  Role of platelet-derived growth factors in physiology and medicine. , 2008, Genes & development.

[74]  M. Sheetz,et al.  Heart extracellular matrix supports cardiomyocyte differentiation of mouse embryonic stem cells. , 2013, Journal of bioscience and bioengineering.

[75]  Karthikeyan Narayanan,et al.  Lineage restricted progenitors for the repopulation of decellularized heart. , 2011, Biomaterials.

[76]  B. Brown,et al.  Chapter II.6.16 – Tissue Engineering with Decellularized Tissues , 2013 .

[77]  K. Hochedlinger,et al.  Induced pluripotency of mouse and human somatic cells. , 2008, Cold Spring Harbor symposia on quantitative biology.

[78]  H Harasaki,et al.  eNOS-overexpressing endothelial cells inhibit platelet aggregation and smooth muscle cell proliferation in vitro. , 2000, Tissue engineering.

[79]  Wolfgang Konertz,et al.  Immune response in patients receiving a bioprosthetic heart valve: lack of response with decellularized valves. , 2011, Tissue engineering. Part A.

[80]  Emmanuel S. Tzanakakis,et al.  Stem cells for heart cell therapies. , 2008, Tissue engineering. Part B, Reviews.

[81]  A. Didangelos,et al.  Proteomics Analysis of Cardiac Extracellular Matrix Remodeling in a Porcine Model of Ischemia/Reperfusion Injury , 2012, Circulation.

[82]  A. Camm,et al.  Left atrial appendage: structure, function, and role in thromboembolism , 1999, Heart.

[83]  D. Navajas,et al.  Effects of freezing/thawing on the mechanical properties of decellularized lungs. , 2013, Journal of biomedical materials research. Part A.

[84]  Colleen M. Witzenburg,et al.  Mechanical changes in the rat right ventricle with decellularization. , 2012, Journal of biomechanics.

[85]  Charles A. Vacanti,et al.  Stimulus-triggered fate conversion of somatic cells into pluripotency , 2014, Nature.

[86]  R. Nerem,et al.  Fluid shear stress alters the hemostatic properties of endothelial outgrowth cells. , 2012, Tissue engineering. Part A.

[87]  Harald C Ott,et al.  Organ engineering based on decellularized matrix scaffolds. , 2011, Trends in molecular medicine.

[88]  Mark M. Davis,et al.  Short-term immunosuppression promotes engraftment of embryonic and induced pluripotent stem cells. , 2011, Cell stem cell.

[89]  Artur Lichtenberg,et al.  Development of a growing rat model for the in vivo assessment of engineered aortic conduits. , 2012, The Journal of surgical research.

[90]  J. Guyette,et al.  Regeneration and Experimental Orthotopic Transplantation of a Bioengineered Kidney , 2013, Nature Medicine.

[91]  Mitsutaka Kadota,et al.  Bidirectional developmental potential in reprogrammed cells with acquired pluripotency , 2014, Nature.

[92]  G. Salama,et al.  High-purity enrichment of functional cardiovascular cells from human iPS cells. , 2012, Cardiovascular research.

[93]  Smadar Cohen,et al.  The effect of sulfation of alginate hydrogels on the specific binding and controlled release of heparin-binding proteins. , 2008, Biomaterials.

[94]  W. Oeveren,et al.  Obstacles in haemocompatibility testing. , 2013 .

[95]  P. Burridge,et al.  A Review of Human Pluripotent Stem Cell-Derived Cardiomyocytes for High-Throughput Drug Discovery, Cardiotoxicity Screening, and Publication Standards , 2013, Journal of Cardiovascular Translational Research.

[96]  Stephen F Badylak,et al.  The extracellular matrix as a scaffold for tissue reconstruction. , 2002, Seminars in cell & developmental biology.

[97]  T. Okano,et al.  Cell sheet engineering for myocardial tissue reconstruction. , 2003, Biomaterials.

[98]  J. Feijen,et al.  Endothelial Cell Seeding on Crosslinked Collagen: Effects of Crosslinking on Endothelial Cell Proliferation and Functional Parameters , 2000, Thrombosis and Haemostasis.

[99]  Ernst Wolner,et al.  The decellularized porcine heart valve matrix in tissue engineering , 2005, Thrombosis and Haemostasis.

[100]  A. Atala,et al.  Decellularization for whole organ bioengineering , 2013, Biomedical materials.

[101]  S. Bellis,et al.  Advantages of RGD peptides for directing cell association with biomaterials. , 2011, Biomaterials.

[102]  M. Hasegawa,et al.  Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome , 2009, Proceedings of the Japan Academy. Series B, Physical and biological sciences.

[103]  H. Kamiya,et al.  Acceleration of autologous in vivo recellularization of decellularized aortic conduits by fibronectin surface coating. , 2013, Biomaterials.

[104]  Christian Schuetz,et al.  Regeneration and orthotopic transplantation of a bioartificial lung , 2010, Nature Medicine.

[105]  S. Andreadis,et al.  Stem cell sources for vascular tissue engineering and regeneration. , 2012, Tissue engineering. Part B, Reviews.

[106]  U. V. von Oppell,et al.  In vitro endothelialization of expanded polytetrafluoroethylene grafts: a clinical case report after 41 months of implantation. , 1997, Journal of vascular surgery.

[107]  Doris A Taylor,et al.  Perfusion-decellularized matrix: using nature's platform to engineer a bioartificial heart , 2008, Nature Medicine.

[108]  Marcelle Machluf,et al.  Acellular cardiac extracellular matrix as a scaffold for tissue engineering: in vitro cell support, remodeling, and biocompatibility. , 2010, Tissue engineering. Part C, Methods.

[109]  X. Ye,et al.  Polyelectrolyte multilayer film on decellularized porcine aortic valve can reduce the adhesion of blood cells without affecting the growth of human circulating progenitor cells. , 2012, Acta biomaterialia.

[110]  Yoshihiro Ito,et al.  A fusion protein of hepatocyte growth factor for immobilization to collagen. , 2007, Biomaterials.

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

[112]  H Reul,et al.  In Vitro Thrombogenicity Testing of Artificial Organs , 1998, The International journal of artificial organs.

[113]  U. Seyfert,et al.  In vitro hemocompatibility testing of biomaterials according to the ISO 10993-4. , 2002, Biomolecular engineering.

[114]  P. McFetridge,et al.  Improved recellularization of ex vivo vascular scaffolds using directed transport gradients to modulate ECM remodeling , 2013, Biotechnology and bioengineering.

[115]  M. Hiles,et al.  Effects of sterilization on an extracellular matrix scaffold: Part II. Bioactivity and matrix interaction , 2007, Journal of materials science. Materials in medicine.

[116]  Hyun Chul Lee,et al.  Remission in models of type 1 diabetes by gene therapy using a single-chain insulin analogue , 2000, Nature.

[117]  T. Waddell,et al.  The effect of decellularization of tracheal allografts on leukocyte infiltration and of recellularization on regulatory T cell recruitment. , 2013, Biomaterials.

[118]  Stephen F Badylak,et al.  An overview of tissue and whole organ decellularization processes. , 2011, Biomaterials.

[119]  Donald O Freytes,et al.  Reprint of: Extracellular matrix as a biological scaffold material: Structure and function. , 2015, Acta biomaterialia.

[120]  G. Daley,et al.  The promise of induced pluripotent stem cells in research and therapy , 2012, Nature.

[121]  M. Murata,et al.  Distinct iPS Cells Show Different Cardiac Differentiation Efficiency , 2013, Stem cells international.

[122]  Ricardo Londono,et al.  Consequences of ineffective decellularization of biologic scaffolds on the host response. , 2012, Biomaterials.

[123]  Ziqiang Yuan,et al.  Cardiac telocytes were decreased during myocardial infarction and their therapeutic effects for ischaemic heart in rat , 2012, Journal of cellular and molecular medicine.

[124]  Daniel J Weiss,et al.  Initial binding and recellularization of decellularized mouse lung scaffolds with bone marrow-derived mesenchymal stromal cells. , 2012, Tissue engineering. Part A.

[125]  Sandeep T. Koshy,et al.  Cardiac Decellularisation With Long-Term Storage and Repopulation With Canine Peripheral Blood Progenitor Cells †,‡ , 2012 .

[126]  Gordana Vunjak-Novakovic,et al.  Cultivation in rotating bioreactors promotes maintenance of cardiac myocyte electrophysiology and molecular properties. , 2003, Tissue engineering.

[127]  Thomas W Gilbert,et al.  Strategies for tissue and organ decellularization , 2012, Journal of cellular biochemistry.

[128]  A. Kajbafzadeh,et al.  Determining the optimal decellularization and sterilization protocol for preparing a tissue scaffold of a human-sized liver tissue. , 2013, Tissue engineering. Part C, Methods.

[129]  Zuping He,et al.  Generation of functional organs from stem cells , 2013, Cell Regeneration.

[130]  Hang Lin,et al.  The effect of collagen-targeting platelet-derived growth factor on cellularization and vascularization of collagen scaffolds. , 2006, Biomaterials.

[131]  Maria-Simonetta Faussone-Pellegrini,et al.  TELOCYTES – a case of serendipity: the winding way from Interstitial Cells of Cajal (ICC), via Interstitial Cajal-Like Cells (ICLC) to TELOCYTES , 2010, Journal of cellular and molecular medicine.

[132]  Michael V Sefton,et al.  Biomaterial-associated thrombosis: roles of coagulation factors, complement, platelets and leukocytes. , 2004, Biomaterials.

[133]  F A Auger,et al.  A completely biological tissue‐engineered human blood vessel , 1998, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[134]  N. Frangogiannis,et al.  The extracellular matrix as a modulator of the inflammatory and reparative response following myocardial infarction. , 2010, Journal of molecular and cellular cardiology.

[135]  Anthony Ratcliffe,et al.  Bioreactors and Bioprocessing for Tissue Engineering , 2002, Annals of the New York Academy of Sciences.

[136]  A. Weiss,et al.  Elastin as a nonthrombogenic biomaterial. , 2011, Tissue engineering. Part B, Reviews.

[137]  Donald O Freytes,et al.  Preparation of cardiac extracellular matrix from an intact porcine heart. , 2010, Tissue engineering. Part C, Methods.

[138]  Ernst Wolner,et al.  Decellularization protocols of porcine heart valves differ importantly in efficiency of cell removal and susceptibility of the matrix to recellularization with human vascular cells. , 2004, The Journal of thoracic and cardiovascular surgery.

[139]  John P McQuilling,et al.  Porcine pancreas extracellular matrix as a platform for endocrine pancreas bioengineering. , 2013, Biomaterials.

[140]  Jörn Hülsmann,et al.  A novel customizable modular bioreactor system for whole-heart cultivation under controlled 3D biomechanical stimulation , 2013, Journal of Artificial Organs.

[141]  T. Gilbert,et al.  Procedure for decellularization of porcine heart by retrograde coronary perfusion. , 2012, Journal of visualized experiments : JoVE.

[142]  W. J. Bos,et al.  An in vitro test model to study the performance and thrombogenicity of cardiovascular devices. , 1989, ASAIO transactions.

[143]  M. Lindsey,et al.  Macrophage roles following myocardial infarction. , 2008, International journal of cardiology.