Abstract
Small intestine submucosa (SIS) has emerged as one of a number of naturally derived extracellular matrix (ECM) biomaterials currently in clinical use. In addition to clinical applications, ECM materials form the basis for a variety of approaches within tissue engineering research. In our preliminary work it was found that SIS can be consistently and reliably made into tubular scaffolds which confer certain potential advantages. Given that decellularization protocols for SIS are applied to sheet-form SIS, it was hypothesized that a tubular-form SIS would behave differently to pre-existing protocols. In this work, tubular SIS was produced and decellularized by the conventional peracetic acid–agitation method, peracetic acid under perfusion along with two commonly used detergent–perfusion protocols. The aim of this was to produce a tubular SIS that was both adequately decellularized and possessing the mechanical properties which would make it a suitable scaffold for oesophageal tissue engineering, which was one of the goals of this work. Analysis was carried out via mechanical tensile testing, DNA quantification, scanning electron and light microscopy, and a metabolic assay, which was used to give an indication of the biocompatibility of each decellularization method. Both peracetic acid protocols were shown to be unsuitable methods with the agitation-protocol-produced SIS, which was poorly decellularized, and the perfusion protocol resulted in poor mechanical properties. Both detergent-based protocols produced well-decellularized SIS, with no adverse mechanical effects; however, one protocol emerged, SDS/Triton X-100, which proved superior in both respects. However, this SIS showed reduced metabolic activity, and this cytotoxic effect was attributed to residual reagents. Consequently, the use of SIS produced using the detergent SD as the decellularization agent was deemed to be the most suitable, although the elimination of the DNase enzyme would give further improvement.
Highlights
The oesophagus as an organ can be affected by a number of medical conditions which may necessitate the need for extensive treatment to correct
The numbers of visible nuclei per square micrometre were found to be far higher in the untreated control SIS (uSIS) control and PAA-2hA groups than in the final stages of sodium dodecyl sulfate (SDS)/TX, sodium deoxycholate (SD)/DN and PAA-12h
The Sodium dodecyl sulfate & Triton X-100 (SDS/TX) group was observed to have the lowest number of histologically visible nuclei per square micrometre of visible tissue by a substantial margin
Summary
The oesophagus as an organ can be affected by a number of medical conditions which may necessitate the need for extensive treatment to correct These conditions can be congenital, such as oesophageal atresia, which is a paediatric condition where the oesophagus forms incorrectly, or alternatively they can be acquired conditions, such as oesophageal cancer [1,2,3]. Successful extracellular matrix (ECM) materials predominantly consist of allogeneic and xenogeneic decellularized tissues Those tissues commonly in use are urinary bladder matrix, skin, pericardium and small intestine submucosa (SIS), which have all been used for a variety of clinical applications and are derived from human, bovine and porcine sources [7].
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