PREPARATION AND BIO-EVALUATION OF TISSUE ENGINEERED SCAFFOLD BASED ON DECELLULARIZED WHOLE HEART EXTRACELLULAR MATRIX_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

1Chinese PLA General Hospital &;
Medical School of Chinese PLA, Beijing, 100853, P.R.China;;
2Department of Advanced Interdisciplinary Studies, Institute of Basic Medical Sciences, Academy of Military Medical Sciences. Corresponding author: WANG Changyong, E-mail: wcy2000@yahoo.com;

Corresponding author：

Keywords：

Cardiac tissue engineering; Decellularized whole heart matrix; Biocharacteristic; Rat

DOI：

10.7507/1002-1892.20130206

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Objective To investigate a method for preparing decellularized rat heart scaffold, and to detect and evaluate the decellularized scaffold. Methods The decellularized rat heart scaffold was prepared by retrograde perfusion with a combination of enzymatic and Triton X-100 detergent methods to remove the populations of resident cells, and then the decellularized scaffold was observed by gross, toluidine blue staining, HE staining, scanning electron microcope (SEM), Alcian blue staining, and immunohistochemisty staining to evaluate the structure and essential component of extracellular maxtix (ECM) in the scaffold. Results Tissue engineered scaffold based on decellularized whole heart ECM was successfully prepared, which maintained not only the gross morphology of the heart, but also the intact vascular structure and ultrastructural conformation that certified by toluidine blue staining, HE staining, and SEM analyses. Alcian blue staining and immunohistochemisty staining showed that the essential components of ECM, such as collagen type I, glycosaminoglycan, fibronectin, and Laminin were remained in decellularized whole heart matrix. Conclusion The decellularized whole heart ECM prepared by method mentioned can maintain the intact structure of rat heart and basic compositions of extracellular matrices, so it could be suitable for further studies of tissue engineered scaffolds for whole heart reconstruction.

Citation： CHEN Weizhen,DUAN Cuimi,LI Xia,SUN Hongyu,MOU Yongchao,SHU Yao CHEN Cheng,FENG Lanlan,ZHOU Jin,WANG Changyong. PREPARATION AND BIO-EVALUATION OF TISSUE ENGINEERED SCAFFOLD BASED ON DECELLULARIZED WHOLE HEART EXTRACELLULAR MATRIX. Chinese Journal of Reparative and Reconstructive Surgery, 2013, 27(8): 945-949. doi: 10.7507/1002-1892.20130206 Copy

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2.	Zimmermann WH, Melnychenko I, Wasmeier G, et al. Engineered heart tissue grafts improve systolic and diastolic function in infarcted rat hearts. Nat Med, 2006, 12(4): 452-458.
3.	Zimmermann WH, Melnychenko I, Eschenhagen T. Engineered heart tissue for regeneration of diseased hearts. Biomaterials, 2004, 25(9): 1639-1647.
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9.	Wainwright JM, Czajka CA, Patel UB, et al. Preparation of cardiac extracellular matrix from an intact porcine heart. Tissue Eng Part C Methods, 2010, 16(3): 525-532.
10.	Akhyari P, Aubin H, Gwanmesia P, et al. The quest for an optimized protocol for whole-heart decellularization: a comparison of three popular and a noveldecellularization technique and their diverse effects on crucial extracellular matrix qualities. Tissue Eng Part C Methods, 2011, 17(9): 915-926.
11.	Tudorache I, Cebotari S, Sturz G, et al. Tissue engineering of heart valves: biomechanical and morphological properties of decellularized heart valves. J Heart Valve Dis, 2007, 16(5): 574-578.
12.	Mol A, Smits AI, Bouten CV, et al. Tissue engineering of heart valves: advances and current challenges. Expert Rev Med Devices, 2009, 6(3): 259-275.
13.	Hoshiba T, Lu H, Kawazoe N, et al. Decellularized matrices for tissue engineering. Expert Opin Biol Ther, 2010, 10(12): 1717-1728.
14.	Turner WS, Wang X, Johnson S, et al. Cardiac tissue development for delivery of embryonic stem cell-derived endothelial and cardiac cells in natural matrices. J Biomed Mater Res B Appl Biomater, 2012, 100(8): 2060-2072.
15.	Ott HC, Clippinger B, Conrad C, et al. Regeneration and orthotopic transplantation of a bioartificial lung. Nat Med, 2010, 16(8): 927-933.
16.	Petersen TH, Calle EA, Zhao L, et al. Tissue-engineered lungs for in vivo implantation. Science, 2010, 329(5991): 538-541.
17.	Uygun BE, Soto-Gutierrez A, Yagi H, et al. Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix. Nat Med, 2010, 16(7): 814-820.
18.	Ross EA, Abrahamson DR, St John P, et al. Mouse stem cells seeded into decellularized rat kidney scaffolds endothelialize and remodel basement membranes. Organogenesis, 2012, 8(2): 49-55.
19.	Sullivan DC, Mirmalek-Sani SH, Deegan DB, et al. Decellularization methods of porcine kidneys for whole organ engineering using a high-throughput system. Biomaterials, 2012, 33(31): 7756-7764.
20.	Nakayama KH, Batchelder CA, Lee CI, et al. Decellularized rhesus monkey kidney as a three-dimensional scaffold for renal tissue engineering. Tissue Eng Part A, 2010, 16(7): 2207-2216.

1. Badylak SF, Taylor D, Uygun K. Whole-organ tissue engineering: decellularization and recellularization of three-dimensional matrix scaffolds. Annu Rev Biomed Eng, 2011, 13(13): 27-53.
2. Zimmermann WH, Melnychenko I, Wasmeier G, et al. Engineered heart tissue grafts improve systolic and diastolic function in infarcted rat hearts. Nat Med, 2006, 12(4): 452-458.
3. Zimmermann WH, Melnychenko I, Eschenhagen T. Engineered heart tissue for regeneration of diseased hearts. Biomaterials, 2004, 25(9): 1639-1647.
4. Song JJ, Ott HC. Organ engineering based on decellularized matrix scaffolds. Trends Mol Med, 2011, 17(8): 424-432.
5. Naito H, Melnychenko I, Didié M, et al. Optimizing engineered heart tissue for therapeutic applications as surrogate heart muscle. Circulation, 2006, 114(1 suppl): 172-178.
6. Gilbert TW, Sellaro TL, Badylak SF. Decellularization of tissues and organs. Biomaterials, 2006, 27(19): 3675-3683.
7. Ott HC, Matthiesen TS, Goh SK, et al. Perfusion-decellularized matrix: using nature’s platform to engineer a bioartificial heart. Nat Med, 2008, 14(2): 213-221.
8. Ng SL, Narayanan K, Gao S, et al. Lineage restricted progenitors for the repopulation of decellularized heart. Biomaterials, 2011, 32(30): 7571-7580.
9. Wainwright JM, Czajka CA, Patel UB, et al. Preparation of cardiac extracellular matrix from an intact porcine heart. Tissue Eng Part C Methods, 2010, 16(3): 525-532.
10. Akhyari P, Aubin H, Gwanmesia P, et al. The quest for an optimized protocol for whole-heart decellularization: a comparison of three popular and a noveldecellularization technique and their diverse effects on crucial extracellular matrix qualities. Tissue Eng Part C Methods, 2011, 17(9): 915-926.
11. Tudorache I, Cebotari S, Sturz G, et al. Tissue engineering of heart valves: biomechanical and morphological properties of decellularized heart valves. J Heart Valve Dis, 2007, 16(5): 574-578.
12. Mol A, Smits AI, Bouten CV, et al. Tissue engineering of heart valves: advances and current challenges. Expert Rev Med Devices, 2009, 6(3): 259-275.
13. Hoshiba T, Lu H, Kawazoe N, et al. Decellularized matrices for tissue engineering. Expert Opin Biol Ther, 2010, 10(12): 1717-1728.
14. Turner WS, Wang X, Johnson S, et al. Cardiac tissue development for delivery of embryonic stem cell-derived endothelial and cardiac cells in natural matrices. J Biomed Mater Res B Appl Biomater, 2012, 100(8): 2060-2072.
15. Ott HC, Clippinger B, Conrad C, et al. Regeneration and orthotopic transplantation of a bioartificial lung. Nat Med, 2010, 16(8): 927-933.
16. Petersen TH, Calle EA, Zhao L, et al. Tissue-engineered lungs for in vivo implantation. Science, 2010, 329(5991): 538-541.
17. Uygun BE, Soto-Gutierrez A, Yagi H, et al. Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix. Nat Med, 2010, 16(7): 814-820.
18. Ross EA, Abrahamson DR, St John P, et al. Mouse stem cells seeded into decellularized rat kidney scaffolds endothelialize and remodel basement membranes. Organogenesis, 2012, 8(2): 49-55.
19. Sullivan DC, Mirmalek-Sani SH, Deegan DB, et al. Decellularization methods of porcine kidneys for whole organ engineering using a high-throughput system. Biomaterials, 2012, 33(31): 7756-7764.
20. Nakayama KH, Batchelder CA, Lee CI, et al. Decellularized rhesus monkey kidney as a three-dimensional scaffold for renal tissue engineering. Tissue Eng Part A, 2010, 16(7): 2207-2216.

Chinese Journal of Reparative and Reconstructive Surgery

PREPARATION AND BIO-EVALUATION OF TISSUE ENGINEERED SCAFFOLD BASED ON DECELLULARIZED WHOLE HEART EXTRACELLULAR MATRIX

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