doi:10.3850/978-981-08-7615-9_TE10

ABSTRACT

Decellularized tissues have shown promising results in tissue/organ reconstruction, mainly due to the bioactivity remaining in the extracellular matrix that initiates tissue regeneration. However, decellularized tissues currently used for esophageal reconstruction still face problems such as (1) stenosis during animal implantation and (2) reliance on enzymes (which are foreign proteins) in some protocols. This paper describes a new chemical-based protocol for decellularizing the porcine esophagus that replaces the use of enzymes for clearing nucleic acids and minimizes structural damage of the decellularized tissue. It involves sequential chemical treatments using EDTA, SDS and a new oxidizing treatment using an iron(III)-ascorbate mixture. Histological staining showed that the decellularized esophageal matrix was nearly devoid of cell nuclei while scanning electron microscopy showed an intact basement membrane with a microstructure that resembled the native esophagus. Quantitatively, a 93% reduction of the DNA content was achieved by the protocol and measurements of collagenase resistance and suture retention strength of the decellularized esophagus were found to be comparable to the native tissue. Furthermore, no in vitro cytotoxicity was observed for the decellularized esophagus. The proposed protocol was able to produce a non-cytotoxic decellularized esophagus with high cellular removal and minimal structural damage. Further in vivo and in vitro studies will allow a more in-depth investigation of the use of the decellularized tissue in esophageal reconstruction.

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