Acellular porcine heart matrices: whole organ decellularization with 3D-bioscaffold & vascular preservation Acellular porcine heart matrices: whole organ decellularization with 3D-bioscaffold & vascular preservation
Regenerative medicine, particularly decellularization-recellularization methods via whole-organ tissue engineering, has been increasingly studied due to the growing donor organ shortage. Though numerous decellularization protocols exist, the ideal decellularization protocol for optimal recellularization is unclear. This study was performed to optimize existing heart decellularization protocols and compare current methods using the detergents SDS (sodium dodecyl sulfate), Triton X-100, OGP (octyl β-D-glucopyranoside), and CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate) through retrograde aortic perfusion via aortic cannulation of a whole porcine heart. The goal of decellularization is to preserve extracellular matrix integrity and architecture, which was analyzed in this study through histology, microscopy, DNA analysis, hydroxyproline content analysis, materials analysis and angiography. Effective decellularization was determined by analyzing the tissue organization, geometry, and biological properties of the resultant extracellular matrix scaffold. Using these parameters, optimal decellularization was achieved between 90 and 120 mmHg pressure with 3% SDS as a detergent.
[1] Tapias LF, Ott HC. Decellularized scaffolds as a platform for bioengineered organs. Curr Opin Organ Transplant 2014; 19: 145–152.
[2] Faulk DM, Johnson SA, Zhang L, Badylak SF. Role of the extracellular matrix in whole organ engineering. J Cell Physiol 2014; 229: 984–989.
[3] Yagi H, Soto-Gutierrez A, Kitagawa Y. Whole-organ re-engineering: a regenerative medicine approach to digestive organ replacement. Surg Today 2013; 43: 587–594.
[4] He M, Callanan A. Comparison of methods for whole-organ decellularization in tissue engineering of bioartificial organs. Tissue Eng Part B Rev 2013; 19: 194–208.
[5] Song JJ, Ott HC. Organ engineering based on decellularized matrix scaffolds. Trends Mol Med 2011; 17: 424–432.
[6] Murphy SV, Atala A. Organ engineering—combining stem cells, biomaterials, and bioreactors to produce bioengineered organs for transplantation. Bioessays 2013; 35: 163–172.
[7] Sullivan DC, Mirmalek-Sani SH, Deegan DB, Baptista PM, Aboushwareb T, Atala A, Yoo JJ. Decellularization methods of porcine kidneys for whole organ engineering using a high-throughput system. Biomaterials 2012; 33: 7756–7764.
[8] Uygun BE, Soto-Gutierrez A, Yagi H, Izamis ML, Guzzardi MA, Shulman C, Milwid J, Kobayashi N, Tilles A, Berthiaume F, Hertl M, Nahmias Y, Yarmush ML, Uygun K. Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix. Nat Med, 2010; 16: 814–820.
[9] Gilpin SE, Guyette JP, Gonzalez G, Ren X, Asara JM, Mathisen DJ, Vacanti JP, Ott HC. Perfusion decellularization of human and porcine lungs: bringing the matrix to clinical scale. J Heart Lung Transplant 2014;33: 298–308.
[10] Jensen T, Roszell B, Zang F, Girard E, Matson A, Thrall R, Jaworski DM, Hatton C, Weiss DJ, Finck C. A rapid lung decellularization protocol supports embryonic stem cell differentiation in vitro and following implantation. Tissue Eng Part C Methods. 2012;18: 632–646.
[11] Khalpey Z, Qu N, Hemphill C, Louis AV, Ferng AS
[12] Orlando G, Baptista P, Birchall M, De Coppi P, Farney A, Guimaraes-Souza NK, Opara E, Rogers J, Seliktar D, Shapira-Schweitzer K, Stratta RJ, Atala A, Wood KJ, Soker S. Regenerative medicine as applied to solid organ transplantation: current status and future challenges. Transpl Int 2011; 24: 223–232.
[13] Tee R, Lokmic Z, Morrison WA, Dilley RJ. Strategies in cardiac tissue engineering. ANZ J Surg 2011; 80: 683–693.
[14] Guyette JP, and Coauthors, 2016: Bioengineering human myocardium on native extracellular matrix. Circ Res 2016; 118: 56–72.
[15] Sánchez PL, Fernández-Santos ME, Costanza S, Climent AM, Moscoso I, Gonzalez-Nicolas MA, Sanz-Ruiz R, Rodríguez H, Kren SM, Garrido G, Escalante JL, BermejoJ, Elizaga J, Menarguez J, Yotti R, Pérez del Villar C, Espinosa MA, Guillem MS, Willerson JT, Bernad A, Matesanz R, Taylor DA, Fernández-Avilés F. Acellular human heart matrix: A critical step toward whole heart grafts. Biomaterials 2015; 61: 279–289.
[16] Oberwallner B, Brodarac A, Choi YH, Saric T, Anić P, Morawietz L, Stamm C. Preparation of cardiac extracellular matrix scaffolds by decellularization of human myocardium. J Biomed Mater Res A 2014; 102: 3263–3272.
[17] Lu TY, Lin B, Kim J, Sullivan M, Tobita K, Salama G, Yang L. Repopulation of decellularized mouse heart with human induced pluripotent stem cell-derived cardiovascular progenitor cells. Nat Commun 2013; 4: 2307.
[18] Ott HC, Matthiesen TS, Goh SK, Black LD, Kren SM, Netoff TI, Taylor DA. Perfusion-decellularized matrix: using nature's platform to engineer a bioartificial heart. Nat Med 2008; 14: 213–221.
[19] Methe K, Bäckdahl H, Johansson BR, Nayakawde N, Dellgren G, Sumitran-Holgersson S. An alternative approach to decellularize whole porcine heart. Biores Open Access 2014; 3: 327–338.
[20] Guyette JP, Gilpin SE, Charest JM, Tapias LF, Ren X, Ott HC. Perfusion decellularization of whole organs. Nat Protoc 2014; 9: 1451–1468.
[21] Choi MY, Kim JT, Lee WJ, Lee Y, Park KM, Yang YI, Park KD. Engineered extracellular microenvironment with a tunable mechanical property for controlling cell behavior and cardiomyogenic fate of cardiac stem cells. Acta Biomater 2017; 50: 234-248
[22] Fernandes H, Moroni L, van Blitterswijk C, de Boer J. Extracellular matrix and tissue engineering applications. Journal of Materials Chemistry 2009; 19: 5474–5484.
[23] Momtahan N, Poornejad N, Struk JA, Castleton AA, Herrod BJ, Vance BR, Eatough JP, Roeder BL, Reynolds PR, Cook AD. Automation of pressure control improves whole porcine heart decellularization. Tissue Eng Part C Methods 2015; 21:1148-1161.
[24] Friedrich LH, Jungebluth P, Sjöqvist S, Lundin V, Haag JC, Lemon G, Gustafsson Y, Ajalloueian F, Sotnichenko A, Kielstein H, Burguillos MA, Joseph B, Teixeira AI, Lim ML, Macchiarini P. Preservation of aortic root architecture and properties using a detergent-enzymatic perfusion protocol. Biomaterials 2014; 35: 1907–1913.
[25] Wainwright JM, Czajka CA, Patel UB, Freytes DO, Tobita K, Gilbert TW, Badylak SF. Preparation of cardiac extracellular matrix from an intact porcine heart. Tissue Eng Part C Methods 2010; 16: 525–532.
[26] Weymann A, Patil NP, Sabashnikov A, Jungebluth P, Korkmaz S, Li S, Veres G, Soos P, Ishtok R, Chaimow N, Pätzold I, Czerny N, Schies C, Schmack B, Popov AF, Simon AR, Karck M, Szabo G. Bioartificial Heart: A Human-Sized Porcine Model - The Way Ahead. PLoS ONE 2014; 9: e111591.
[27] Petersen TH, Calle EA, Colehour MB, Niklason LE. Matrix composition and mechanics of decellularized lung scaffolds. Cells Tissues Organs (Print) 2012; 195: 222–231.
[28] Remlinger NT, Wearden PD, Gilbert TW. Procedure for decellularization of porcine heart by retrograde coronary perfusion. J Vis Exp 2012: e50059.
[29] Booth C, Korossis SA, Wilcox HE, Watterson KG, Kearney JN, Fisher J, Ingham E. Tissue engineering of cardiac valve prostheses I: Development and histological characterization of an acellular porcine scaffold. J Heart Valve Dis 2002; 11: 457–462.
[30] Dong J, Li Y, Mo X. The study of a new detergent (octyl-glucopyranoside) for decellularizing porcine pericardium as tissue engineering scaffold. J Surg Res 2013; 183: 56–67.
[31] Crapo PM, Gilbert TW, Badylak SF. An overview of tissue and whole organ decellularization processes. Biomaterials 2011; 32: 3233–3243.
[32] Nagata S, Hanayama R, Kawane K. Autoimmunity and the clearance of dead cells. Cell 2010; 140: 619–630.