Development of an Esophagus Acellular Matrix Tissue Scaffold

Abstract

A cell-extraction protocol yielding an esophagus acellular matrix (EAM) scaffold for use in tissue engineering of an esophagus, including hypotonic lysis, multiple detergent cell extraction steps, and nucleic acid digestion, was developed in a rat model. Histological techniques, burst pressure studies, in vitro esophageal epithelial cell seeding, and in vivo implantation were used to assess cell extraction, extracellular matrix (ECM) preservation, and biocompatibility. Microscopy demonstrated that cell extraction protocols using sodium dodecyl sulfate (SDS) (0.5%, wt/vol) as a detergent resulted in cell-free EAM with retained ECM protein collagen, elastin, laminin, and fibronectin. Burst pressure studies indicated a loss of tensile strength in EAMs, but at intraluminal pressures that were unlikely to affect in vivo application. In vitro cell seeding studies exhibited epithelial cell proliferation with stratification similar to native esophagi after 11 days, and subcutaneously implanted EAMs displayed neovascularization and a minimal inflammatory response after 30 days of implantation. This study presents an esophagus acellular matrix tissue scaffold with preserved ECM proteins, biomechanical properties, and the ability to support esophageal cell proliferation to serve as the foundation for a tissue-engineered esophagus.

Mouse stem cells seeded into decellularized rat kidney scaffolds endothelialize and remodel basement membranes

“…[4][5][6][7][8] We have had concerns, however, that simple applications of lineage-specific (i.e., endothelial or chondrocyte) cells would not be adequate for complex organs. Kidney generation, for example, would require multiple types of cells in intricate three dimensional heterogeneous arrangements.…”

“…Rather than de novo organ growth, [1][2][3] there has been recent successful repopulation of acellular scaffolds derived from a variety of nonrenal tissues. [4][5][6][7][8] At issue, however, has been how to seed decellularized complex organs with mature or precursor cells so as to reproduce intricate architectures: multiple cell types properly configured and with appropriate cell-to-cell alignment. We have hypothesized that properly prepared scaffolds retain extracellular matrix constituents that can signal pluripotent precursor cells for appropriate site-specific differentiation.…”

Tissue Engineering of Esophagus

“…These natural materials also have been used in a variety of tissue engineering applications, such as the grafts of heart, heart valve, skeletal muscle, skin, cardiovascular grafts, etc. For the research of esophageal tissue engineering, some ECMs like acellular dermal grafts, gastric acellular matrix, aortal acellular matrix grafts, and decellularized esophagus and urinary bladder submucosa had once been tested to repair esophagus in animal models [11][12][13][14]. For example, Marzaro et al seeded porcine esophageal smooth muscle cells (SMC) on the acellular esophagus aiming at healing the defected porcine esophagus.…”

Whole-organ engineering with natural extracellular materials

“…These scaffolds retain the original architecture of organs and tissue interfaces, giving rise to a source of allogenic and xenogenic whole--organ grafts. Decellularization has been carried out on a variety of tissues of various organs such as the urinary bladder, small intestinal submucosa, blood vessels, heart valves, pericardium, tooth buds, trachea and esophagus, as well as musculoskeletal regions such as the temporomandibular joint [9][10][11][12][13][14][15][16][17][18][19][20][21][22]. This method of creating acellular scaffolds has resulted in commercial products and tissue substitutes [23][24][25][26][27].…”

“…Other imaging techniques such as scanning-electron microscopy have been used to visualize and compare nanostructures (e.g., the weave, coil and strut fibers in the decellularized heart study) in decellularized and non-decellularized cadaveric tissue [32]. Together, these studies show that with optimal treatment, decellularized tissues can maintain ECM antibody epitope expression in such molecules as collagens, laminin and fibronectin in immunohistochemistry and immunofluorescence studies [13,15,18,21,30,36]. Table 2 summarizes the various ECM molecules characterized in select decellularized organ and tissue studies.…”