Effect of bovine pericardial extracellular matrix scaffold niche on seeded human mesenchymal stem cell function

Abstract

Numerous studies have focused on generation of unfixed bovine pericardium (BP) extracellular matrix (ECM) for clinical application. However, the extent to which maintenance of native ECM niche is capable of directing behavior of repopulating cells remains relatively unexplored. By exploiting the sidedness of BP scaffolds (i.e., serous or fibrous surface), this study aims to determine the effect of ECM niche preservation on cellular repopulation using different scaffold generation methods. BP underwent either sodium dodecyl sulfate (SDS) decellularization or stepwise, solubilization-based antigen removal using amidosulfobetaine-14 (ASB-14). SDS scaffolds were toxic to repopulating human mesenchymal stem cells (hMSC). Scanning electron microscopy revealed distinct surface ultrastructure of ASB-14 scaffolds based on native BP sidedness. Basement membrane structures on the serous side stimulated hMSC cell monolayer formation, whereas fibrous side facilitated cell penetration into scaffold. Additionally, serous side seeding significantly increased hMSC adhesion and proliferation rate compared to the fibrous side. Furthermore, scaffold ECM niche stimulated sidedness dependent differential hMSC human leukocyte antigen expression, angiogenic and inflammatory cytokine secretion. This work demonstrates that ECM scaffold preparation method and preservation of BP side-based niches critically affects in vitro cell growth patterns and behavior, which has implications for use of such ECM biomaterials in clinical practice.

In vitro evaluation of bovine pericardium after a soft decellularization approach for use in tissue engineering

“…Prior to the initiation of the biocompatibility assays of the BPs, it is necessary to be aware of tissue cytotoxicity after the decellularization process. In this context, a residual cytotoxic effect, attributed to the strong anionic detergent potential, was previously demonstrated in SDS‐treated tissues even after 7 days of washing . High levels of detergent residues are related to loss of tensile strength and to increased elastin fiber proteolysis, which may lead to subsequent structural degeneration .…”

“…The use of hASCs for tissue repopulation is desirable mainly due to their immunomodulatory and pro‐regenerative properties . In addition, hASCs can be obtained from the patient without an invasive removal procedure, allowing a humanized and autologous transplant.…”

Graft-specific immune tolerance is determined by residual antigenicity of xenogeneic extracellular matrix scaffolds

“…This results in persistence of both known (e.g., MHC I) and unknown (i.e., 70% residual antigenicity remaining) lipophilic antigens [3,13,32]. Furthermore, studies have shown SDS to severely denature vital ECM proteins and be toxic to repopulating cells, stimulating recipient innate immune mediated foreign body response [3,10,13,33–35]. Conversely, zwitterionic detergents (e.g., sulfobetaines) are powerful protein solubilizers due to their hydrophilic head and hydrophobic tail [23,24,36].…”

Application of tissue‐engineered pericardial patch in rat models of myocardial infarction

“…To ameliorate stem cell engraftment, several approaches have been investigated including genetic manipulation of cells to increase homing and survival as well as tissue engineering techniques involving preparation of scaffolds . Scaffolds that are derived from decellularized natural tissues appear to be appropriate and efficient as they possess the essential natural tunnels for vessels and nerves as well as the porosity to nest‐specific cell lineage.…”

“…15 To ameliorate stem cell engraftment, several approaches have been investigated including genetic manipulation of cells to increase homing and survival as well as tissue engineering techniques involving preparation of scaffolds. 5,[16][17][18][19] Scaffolds that are derived from decellularized natural tissues appear to be appropriate and efficient as they possess the essential natural tunnels for vessels and nerves as well as the porosity to nest-specific cell lineage. An appropriate approach for decellularization of scaffolds would preserve the extracellular matrices' integrity and structural features resulting in better signaling and cell homage, while at the same time omitting cells and reducing immunologic reactions.…”