Comparative local immunogenic potential of scaffolds prepared from porcine cholecyst, jejunum, and urinary bladder in rat subcutaneous model

Abstract

Extracellular matrices isolated from several mammalian organs/tissues have found several clinical uses as xenografts or implants. However, they may cause complications because of adverse immunologic reactions. Scaffolds that promote favorable graft-acceptance reaction are preferred for fabricating xenografts. The objective of this study was to evaluate the immunogenic potential of a porcine cholecyst-derived scaffold (CDS), prepared by a non-detergent/enzymatic method, in comparison with jejunum and urinary bladder-derived scaffolds in a rat subcutaneous model. Key graft-rejection/acceptance reaction was evaluated at the site of implantation by studying the occurrence and/or function of immunocompetent cells in the tissue reaction. There was differential occurrence of M1-macrophage, M2-macrophage, T-helper cells, T-cytotoxic cells, B-cells, and mast cells in the tissue reaction and the CDS attracted few cells compared with other scaffolds. Real-time polymerase chain reaction for evaluating mRNA of functional markers like inducible nitric oxide synthase (M1 macrophage), arginase 1 (M2 macrophage), interferon gamma (TH1 lymphocytes), and interlukin-4 (TH2 lymphocytes) suggested that the CDS, compared with the scaffolds prepared from small intestine and urinary bladder, elicited M2 macrophage and TH2 lymphocyte polarization that are congenial graft-acceptance reactions. The results indicated that CDS has less immunogenic potential compared with the scaffolds prepared from jejunum and urinary bladder when used as subcutaneous graft in rats. It was concluded that CDS is a promising animal-derived xenograft for biomedical application.

Comparative profiling of extractable proteins in extracellular matrices of porcine cholecyst and jejunum intended for preparation of tissue engineering scaffolds

“…The CDE contained at least 154 proteins, whereas JDE had 186 proteins indicating the diversity of detectable proteins in the respective extracts. Of these, 79% proteins in CDE and 90% proteins in JDE were of cellular in origin (Table ), possibly indicating the rarity of cell debris in CDE . The finding is not surprising and it is known that even after rigorous decellularization procedures involving, detergents, enzymes, and/or decontaminating agents are not expected to cause complete removal of cellular residue .…”

“…Despite these drawbacks, the study indicated the relative absence of immunogenic proteins in the ECM of cholecyst that can cause adverse host reactions. This sparse content of protein load might have contributed for the lower immunogenic potential of CDS in comparison with scaffolds prepared from jejunum …”

“…Whereas the method of preparation significantly affects the quality and biomolecular composition of a scaffold, the source organ and donor species may also contribute for the same. Exploratory studies have indicated that, when prepared by the same method, porcine CDS has lower immunogenic potential compared to scaffolds prepared from jejunum of the same species . Therefore, it is reasonable to hypothesize that the differential property of CDS is due to the protein composition of the tissue of origin.…”

“…Exploratory studies have indicated that, when prepared by the same method, porcine CDS has lower immunogenic potential compared to scaffolds prepared from jejunum of the same species. 15 Therefore, it is reasonable to hypothesize that the differential property of CDS is due to the protein composition of the tissue of origin. Against this background, this study investigated the nature of extractable protein composition of ECMs of porcine cholecyst and jejunum, which are two favorite source organs for scaffold preparation.…”

Controlled cross‐linking of porcine cholecyst extracellular matrix for preparing tissue engineering scaffold

“…The method has been used for preparing tissue engineering scaffolds from porcine cholecyst (gall bladder), jejunum (small intestine), and urinary bladder. When these scaffolds were used as a subcutaneous graft (Muhamed, Revi, Rajan, & Anilkumar, ) or as a skeletal muscle graft (Balakrishnan‐Nair et al, ), the scaffold prepared from cholecyst known as cholecyst derived scaffold (CDS) promoted an anti‐inflammatory tissue remodeling response rather than a graft rejection reaction in animal models, indicating the possibility of using the CDS as scaffolds for fabricating tissue constructs. On the other hand, the bioinductive properties of any material are known to compromise with exposure to higher concentration of cross‐linking agent (Sart et al, ).…”