Development of Decellularized Xenogeneic Scaffold for Cardiac Tissue Engineering Applications

Abstract

Recent advancements in tissue engineering have paved the way to fabricate varieties of scaffolds from xenogeneic sources to provide improvement in cardiovascular applications. The main objective of the study was to develop the scaffold from a xenogeneic source and explore the possibility of using caprine pericardium in the field of tissue engineering applications. The first part of the study focuses on the optimization of the decellularization of the caprine pericardium with various concentrations of a non-ionic surfactant and anionic detergent. Three different decellularization procedures based on anionic detergent, non-ionic surfactant, and a combination of both were used to achieve the decellularized caprine pericardium (DCP). The efficiency of decellularization protocol was assessed by protein estimation, histology, and DNA quantification. Further, to identify the impact of decellularization on extracellular matrix (ECM) damage, the biomechanical properties of the native pericardium and DCP were investigated. The result showed that optimal concentration and combinations of anionic detergent and non-ionic surfactants play a significant role in gaining decellularized tissues with intact ECM. In the next part of the study, hemocompatibility and physicochemical properties of the DCP were obtained from the optimized concentration of anionic detergent and non-ionic surfactant. were explored. Hemolysis, thrombotic, and platelet assay showed that the DCP was compatible with blood. The successful sterilization of DCP was obtained from antibiotics/antimycotic treatment when compared to the ethanol sterilization technique. Next part of the study focused on analyzing the biological interaction of the serous and fibrous side of the DCP with valvular interstitial cells (VIC) and mimicking the aortic valve calcification process by recellularization with VIC. The biocompatibility studies show that the DCP was capable of supporting cell binding, adhesion, and proliferation of VIC. In particular, the serous side of DCP promotes cell binding and proliferation, and the fibrous side supports cell infiltration. The aortic valve calcification process was mimicked in the in vitro conditions by changing the medium with the different concentrations of calcium and phosphate with DCP. The atomic absorption spectroscopy and alizarin red staining results show that the combination of calcium and phosphate has a critical role in the accelerated calcification of DCP. Final part of the study was intended to develop a biohybrid scaffold of DCP and graphene oxide (GO) by an immersion coating technique. Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, and scanning electron microscope (SEM) were performed to characterize GO-DCP. The presence of GO on the surface of the GO-DCP biohybrid was confirmed by SEM analysis. The existence of glycosaminoglycan, elastin, and collagen in the DCP and GO-DCP was inferred from the FTIR. The antimicrobial activity of GO was evaluated against E. coli and showed minimum inhibitory concentration at 125 μg/ml and minimum bactericidal concentration at 250 μg/ml. The biocompatibility of GO-DCP results shows that GO coating supports cell adhesion on the serous and fibrous sides of the DCP. Further, the biomechanical response of DCP was unaltered by the presence of GO. Thus, the scaffold developed from the caprine model was explored in this study, and the result indicates that the DCP is hemocompatible, noncytotoxic, and biocompatible. These properties indicate that the caprine pericardium can be utilized for cardiac tissue engineering applications.