Decellularization of porcine carotid by the recipient’s serum and evaluation of its biocompatibility using a rat autograft model

Abstract

Recently, decellularized tissues for organ transplantation and regeneration have been actively studied in the field of tissue engineering. In the decellularization process, surfactants such as sodium dodecyl sulfate (SDS) have been most commonly used to remove cellular components from the tissue. However, the residual surfactant may be cytotoxic in vivo and has been reported to hinder remodeling after implantation. In addition, treatment with surfactants may destroy the important extracellular matrix (ECM) structure that allows the decellularized tissue to function as a scaffold for cells. In this study, decellularized tissues with high biocompatibility were created using the recipient's serum. By immersing a heterogeneous tissue in serum conditioned to activate the complement system and DNase I, its cellular components could be removed. Compared to an SDS-treated graft, the serum-treated graft preserved the native structure of its ECM. When subcutaneously implanted into an isogenic inbred rat, the graft treated with the recipient's serum resulted in less immunorejection than did the SDS-treated graft.

Limitations of recellularized biological scaffolds for human transplantation

“…However, it has been shown that even after intense washings of an SDS‐decellularized scaffold, seeded cells do not penetrate and attach (Gratzer et al, ), suggesting that factors other than solely entrapped SDS contribute to the poor engraftment of the cells. SDS has been reported to alter ECM composition, and this modification to the native architecture of the scaffold is likely another cause of reduced recellularization (Gratzer et al, ; Ishino & Fujisato, ; Nichols et al, ; Wong et al, ). Despite the deleterious effects of this detergent, there are reports showing that decellularization with SDS results in a recellularized and functional organ (Song et al, ).…”

Revascularization of decellularized lung scaffolds: principles and progress

“…Specifically, we suggest that some of methodologies used to decellularize large-and small-diameter peripheral blood vessels might also be applied for the vascular compartment of whole lungs. In the periphery, these methods yielded excellent preservation of ECM in, e.g., bovine femoral arteries (24), ovine carotid arteries (59), porcine aortic roots (75), and porcine carotid arteries (38). Tissue-engineered vascular grafts made from decellularized peripheral blood vessels have the benefit of containing native vascular ECM proteins but must still overcome problems of mechanical strength and immunogenicity (65).…”

“…Tissue-engineered vascular grafts made from decellularized peripheral blood vessels have the benefit of containing native vascular ECM proteins but must still overcome problems of mechanical strength and immunogenicity (65). In some of these studies, decellularization with either 0.1% SDS, autologous/allogeneic serum (i.e., endogenous proteolytic activity from the complement system and endonucleases), or 1% TX resulted in differential preservation of ECM components, higher elastic moduli, and lower compliance of the decellularized vascular grafts compared with native vessels (24,38,59,75). Preservation and/or ease of restoration of the elastance and compliance of the vascular compartment in the lung will have to be integral considerations for future pulmonary decellularization protocols.…”

“…Instead, limiting the concentration of detergents in "whole lung decellularization solutions," while maintaining physiological perfusion pressures, and/or use of autologous serum, as described with porcine carotid arteries (38), may also be beneficial for preserving ECM composition in the pulmonary vasculature. Using low concentrations of SDS as the main decellularization agent has shown good maintenance of collagens as well as GAGs in decellularized human common femoral arteries (128).…”

“…In addition, maintaining physiological perfusion pressure will be critical for preserving collagens and EN within the pulmonary vascular tree of bioengineered DLS. Future protocols may also consider significantly shortening the time of detergent exposure, e.g., using intermittent flow regimen (58) in combination with the use of protease inhibitors (128) and/or autologous serum (38), to decrease the loss of critical matrix components within the pulmonary vasculature.…”