Abstract
Background
In cardiovascular surgical practice, there is requisite for conduits and patches both in congenital and acquired heart diseases. There are several important limitations in the commercially available grafts apart from their cost factor. The mismatch between the availability vs. need of homografts, have led to processed xenografts to develop into important substitutes. The decellularization process remains crucial, as the presence of cells implies adverse immune response.
Methods
Bovine pericardium and porcine pulmonary artery were harvested and procured from an inspected abattoir under sterile conditions were processed. Acellularity and elastic fiber orientation of the processed tissues was proven by Haematoxylin and Eosin staining and Elastic van Geison staining (EVG). Immuno histochemistry studies were utilized to detect the presence of essential extracellular matrix molecules such as collagen type I, collagen type IV, laminin, fibronectin in the processed xenografts (bovine pericardium, porcine pulmonary artery) along with total Glycosaminoglycans (GAG’s) quantification by Dimethylmethylene Blue (DMMB) assay.
Results and conclusion
It was found that processed xenografts were completely acellular but the extracellular matrix proteins collagen type I, collagen type IV, laminin, fibronectin and total GAG’s were preserved and not damaged by the procedure. Thus enabling us to explain their promise of autologous cell deposition in a span of 6 months (large animal experimental studies).
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References
Chen QZ, Harding SE, Ali NN, Lyon AR, Boccaccini AR. Biomaterials in cardiac tissue engineering: ten years of research survey. Mater Sci Eng R. 2008;59:1–37.
Lee WK, Park KD, Han DK, Suh H, Park JC, Kim YH. Heparinized bovine pericardium as a novel cardiovascular bioprosthesis. Biomaterials. 2000;21:23–30.
Booth C, Korossis SA, Wilcox HE, Watterson KG, Kearney JN, Fisher J, et al. Tissue engineering of cardiac valve prostheses I (development and histological characterization of an acellular porcine scaffold). J Heart Valve Dis. 2002;11:457–62.
Gilbert TW, Sellaro TL, Badylak SF. Decellularization of tissues and organs. Biomaterials. 2006;27:3675–83.
Badylak SF. Xenogeneic extracellular matrix as a scaffold for tissue reconstruction. Transpl Immunol. 2004;12:367–77.
Schenke-Layland K, Vasilevski O, Opitz F, König K, Riemann I, Halbhuber KJ, et al. Impact of decellularization of xenogeneic tissue on extracellular matrix integrity for tissue engineering of heart valves. J Struct Biol. 2003;143:201–8.
Lichtenberg A, Tudorache I, Cebotari S, et al. In vitro re-endothelialization of detergent decellularized heart valves under simulated physiological dynamic conditions. Biomaterials. 2006;27:4221–9.
Guhathakurta S, Varghese S, Balasubramanian V, Agarwal R, Murthy BS, Veerappan S, et al. Technique to process xenogenic tissues for cardiovascular implantation—a preliminary report. Curr Sci. 2006;91:1068–73.
Farndale RW, Buttle DJ, Barrett AJ. Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim Biophys Acta. 1986;883:173–7.
Liao J, Joyce EM, Sacks MS. Effects of decellularization on the mechanical and structural properties of the porcine aortic valve leaflet. Biomaterials. 2008;29:1065–74.
Grauss RW, Hazekamp MG, Oppenhuizen F, Van Munsteren CJ, Gittenberger-De Groot AC, Deruiter MC. Histological evaluation of decellularised porcine aortic valves: matrix changes due to different decellularisation methods. Eur J Cardiothorac Surg. 2005;27(4):566–71.
Guhathakurta S, Balasubramanian V, Ananthakrishnan B, Veerappan S, Balasundari R, Tata BVR, et al. Thrombogenicity studies of three different variants of processed bovine pericardium. IRBM. 2008;29:223–30.
Juthier F, Vincentelli A, Gaudric J. Decellularized heart valve as a scaffold for in vivo recellularization: deleterious effects of granulocyte colony-stimulating factor. J Thorac Cardiovasc Surg. 2006;131:843–52.
Vander Rest M, Garrone R. Collagen family of proteins. FASEB J. 1991;5:2814–23.
Barnard K, Gathercole LJ. Short and long range order in basement membrane type IV collagen revealed by enzymic and chemical extraction. Int J Biol Macromol. 1991;13:359–65.
Miyamoto S, Katz BZ, Lafrenie RM, Yamada KM. Fibronectin and integrins in cell adhesion, signaling and morphogenesis. Ann NY Acad Sci. 1998;857:119–29.
Rosso F, Giordano A, Barbarisi M, Barbarisi A. From cell-ECM interactions to tissue engineering. J Cell Physiol. 2004;199:174–80.
Brown B, Lindberg K, Reing J, Stolz DB, Badylak SF. The basement membrane component of biologic scaffolds derived from extracellular matrix. Tissue Eng. 2006;12:519–26.
Lovekamp JJ, Simionescu DT, Mercuri JJ, Zubiate B, Sacks MS, Vyavahare NR. Stability and function of glycosaminoglycans in porcine bioprosthetic heart valves. Biomaterials. 2006;27(8):1507–18.
Neame PJ, Barry FP. The link proteins. EXS. 1994;70:53–72.
Ramesh B, Ruchi G, Veerappan S, Radha C, Sarasabharathi A, Soma Guhathakurta S, et al. Complete microbe free processed porcine xenograft for clinical use. J Thorac Cardiovasc Surg. 2007;23:240–5.
Acknowledgements
The authors thank Mr. Janardhan Reddy for preparing immunohistochemical sections, R. Balasundari and Sheerin Begam Nasser for assistance with decellularization, Dillip Kumar Bishi, Suneel Rallapalli for their scientific inputs and Pandian A. for technical backing.
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Galla, S., Mathapati, S., Nayak, V.M. et al. Analytical study to evaluate the extracellular matrix in processed acellular xenografts. Indian J Thorac Cardiovasc Surg 26, 132–138 (2010). https://doi.org/10.1007/s12055-010-0026-8
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DOI: https://doi.org/10.1007/s12055-010-0026-8