Comparative profiling of extractable proteins in extracellular matrices of porcine cholecyst and jejunum intended for preparation of tissue engineering scaffolds Jaseer Muhamed, 1 Akhila Rajan, 1 Arun Surendran, 2 Abdul Jaleel, 2 Thapasimuthu V. Anilkumar 1 1 Division of Experimental Pathology, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram, Kerala, India 2 Department of Cardiovascular & Diabetes Disease Biology, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala, India Received 21 June 2015; revised 7 October 2015; accepted 21 October 2015 Published online 6 November 2015 in Wiley Online Library (wileyonlinelibrary.com). DOI: 10.1002/jbm.b.33567 Abstract: Scaffolds prepared from cholecyst and jejunum have differential immunological potential, despite similar biocom- patibility, when used as subcutaneous grafts. The reason for differential immunogenicity is probably due to differences in the nature of protein composition and biomolecules in the extracellular matrices (ECMs) of source organs that are used for preparation of the scaffolds. Against this background, the present study aims to identify the extractable proteins of ECMs derived from porcine cholecyst and jejunum. The proteins were extracted and identified through a conventional database search following sodium dodecyl sulfate-polyacrylamide gel- electrophoresis separation and mass spectroscopy. The result- ant protein profile was analyzed and at least 154 proteins in cholecyst-derived extracellular matrix (CDE) and 186 proteins in jejunum-derived extracellular matrix (JDE) were identified. Both the matrices contained several extracelluar proteins including fibronectin, nidogen, decorin, and lumican that are known to participate in wound healing responses. However, the CDE had fewer cellular proteins than JDE, especially the lat- ter contained class-I and class-II histocompatibility antigens which are incriminated as potent immunogens responsible for graft rejection. The results of the study suggested that the ECMs used for the scaffold preparation need not be “acellular” and differences in the protein composition of the ECMs might have caused the differential wound healing responses. V C 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 489–496, 2017. Key Words: xenograft, extracellular matrix, cholecyst, scaffold, protein profile How to cite this article: Muhamed J, Rajan A, Surendran A, Jaleel A, Anilkumar TV. 2017. Comparative profiling of extractable proteins in extracellular matrices of porcine cholecyst and jejunum intended for preparation of tissue engineering scaffolds. J Biomed Mater Res Part B 2017:105B:489–496. INTRODUCTION Biologic scaffolds prepared from porcine organs/tissues have been used for fabricating wound healing matrix, 1 hernia repair mesh, 2 pubovaginal sling, 3 dural graft, 4 and nerve guidance conduits 5 for therapeutic purposes. These scaffolds, suitable for xenografting, are essentially extracellular matrices (ECMs) of variable purity resulting from appropriate scaffold preparation method. 6 More than a million patients have ben- efitted with such graft procedures. 7 Unlike the synthetic bio- materials, the success of such animal-derived materials depended on their ability to induce site-specific constructive tissue remodeling at the graft site which in turn depends on their inherent biomolecular composition. In fact, biological scaffolds contain variable extent of biomolecules like growth factors that promote tissue regeneration and wound heal- ing. 7,8 However, only a few studies have examined the protein profile of ECMs used for the preparation of animal-derived scaffolds. A protein profiling study of xenograft derived from urinary bladder submucosa showed the presence of several proteins in the matrix that might be the reason to promote wound healing and tissue remodeling response when it was used for regenerative applications. 9 There is no literature available on the protein profile of ECMs scaffolds derived from porcine hollow organs, other than urinary bladder submucosa. Cholecyst-derived scaffold (CDS) prepared from porcine cholecyst (gall bladder) ECM is a promising biological scaffold material that has got potential application in wound healing, 10 urinary bladder reconstruction, 11 and staple line reinforcement during gut surgery. 12 It has been reported that, when prepared by a nondetergent/enzymatic method, 13 CDS has differential immunogenic potential in rat compared to commercial grade scaffolds derived from small intestinal submucosa. 14 On the other hand, the CDS induced faster healing of excision wounds in animal models. 10 Certainly, the differential function of CDS probably reflects its biomolecular composition. Correspondence to: T. V. Anilkumar; e-mail: tvanilkumar@sctimst.ac.in V C 2015 WILEY PERIODICALS, INC. 489