Evaluation of decellularized esophagus as a scaffold for cultured esophageal epithelial cells

Abstract

Recently, decellularized tissue has been reported to have the potential to regenerate a variety of tissues. However, the optimal protocol for a decellularized esophagus has not been studied. Here, we investigated the effect of different decellularization protocols on the histology and biocompatibility of decellularized esophagi in view of future applications to tissue engineering. The esophageal mucosal epithelium (EP) from 4-week-old Wistar rats was enzymatically dissociated and cultured with growth-arrested feeder cells. Two methods for decellularization using deoxycholic acid (DEOX) or Triton X-100 (TRITON) were compared on esophagi from adult Wistar rats. Those treated with DEOX showed superior mechanical properties, maintenance of extracellular matrix, and lower DNA content than those treated with TRITON. To evaluate the biocompatibility of the scaffold, cultured (passage 3) esophageal epithelial cells were seeded inside the decellularized esophagus and cultured for 7 days. The cells seeded onto the decellularized esophagus were examined histologically and immunocytochemically. Esophageal epithelial cells were stratified into three to four cellular layers in vitro inside the decellularized esophagus, to show polarity. The results from immunocytochemistry indicated that the seeded epithelial cells expressed characteristic marker proteins for native esophageal EP. Decellularized esophagus showed suitable compatibility as a scaffold material for esophageal tissue engineering.

Preparation of Cardiac Extracellular Matrix from an Intact Porcine Heart

“…13,14 Convincing arguments can be made for the advantages of tissue-specific ECM scaffolds for tissue-specific repair. [15][16][17][18][19][20][21][22] Logically, a biologic scaffold derived from the targeted tissue source would possess the ideal threedimensional (3D) architecture and biochemical composition to support tissue-specific cell phenotype, cell proliferation, and tissue biomechanical properties. If organs can be decellularized and still maintain their 3D integrity, the resulting scaffold would possibly represent the ideal scaffold for all components of the organ, including vascular and lymphatic structures, nerves, and the parenchymal cells.…”

Chemical Decellularization Methods and Its Effects on Extracellular Matrix

“…One of the reasons for using hypotonic solutions is a minimal change in the structure of the ECM (50). Moreover, hypotonic and hypertonic solutions are used for washing cell debris, but they are not able to clean and remove cell debris from the tissues entirely (51)(52)(53).…”

“…These chemical's compositions destruct cell connections to collagen and fibronectin by separating ions such as Ca 2+ and Mg 2 (97-100). Chelating agents are usually used in combination with enzymes such as trypsin (9,52,(101)(102)(103) or detergents (9,101) and do not have any application on its own (104). EDTA in the long term reduces the mechanical properties of the scaffold (105).…”

Esophageal tissue engineering

“…Ozeki et al ont comparé ces méthodes et concluaient à la supériorité du DEOX au Triton X-100. Le DEOX permettait une diminution plus importante de la quantité d'ADN résiduels [78]. Les procédés de décellularisation permettent de réduire l'inflammation, et la calcification du scaffold après implantation parce qu'il y a parallèlement à la décellularisation une réduction du Complexe Majeure d'Histocompatibilité (CMH) [79].…”

“…Les cellules utilisées peuvent être différenciées ou « à potentiel souches ». En ingénierie tissulaire oesophagienne [133], les cellules utilisées étaient principalement différenciées et représentées par des cellules épithéliales [78,82,83], des cellules musculaires lisses [84,89], des cellules musculaires striées [86,134] et des fibroblastes [64]. L'utilisation de cellules souches a été décrite dans l'ingénierie tissulaire oesophagienne [93].…”