Scaffold with a natural mesh-like architecture: Isolation, structural, and in vitro characterization

An intact extracellular matrix (ECM) with a mesh-like architecture has been identified in the peri-muscular sub-serosal connective tissue (PSCT) of cholecyst (gallbladder). The PSCT layer of cholecyst wall is isolated by mechanical delamination of other layers and decellularized with a treatment with peracetic acid and ethanol solution (PES) in water to obtain the final matrix, which is referred to as cholecyst-derived ECM (CEM). CEM is cross-linked with different concentrations of glutaraldehyde (GA) to demonstrate that the susceptibility of CEM to degradation can be controlled. Quantitative and qualitative macromolecular composition assessments revealed that collagen is the primary structural component of CEM. Elastin is also present. In addition, the ultra-structural studies on CEM reveal the presence of a three-dimensional fibrous mesh-like network structure with similar nanoscale architecture on both mucosal and serosal surfaces. In vitro cell culture studies show that CEM provides a supporting structure for the attachment and proliferation of murine fibroblasts (3T3) and human umbilical vein endothelial cells (HUVEC). CEM is also shown to support the attachment and differentiation of rat adrenal pheochromocytoma cells (PC12).