Effectiveness of decellularisation & extraction of surfactant in porcine oesophagus

Abstract

Background: A decellularised oesophageal scaffold is utilised to replace malignant oesophagus tissue with biological scaffolds. The best tissue engineered construct is highly dependent on the efficacy of the removal of the cellular material within the scaffold material.

Methods: Porcine oesophagi were decellularised by soaking in 0.25% sodium dodecyl sulfate (SDS) at different flow rates within the shortest period of time. Efficiency of decellularisation is then assessed by histology and DNA quantitation. For further analysis to determine the effectiveness of decellularisation, SDS quantification method is used to determine the content of the SDS in the decellularised tissue before extraction of the chemical with distilled water, 1% and 5% ethanol, to ensure that the tissue have minimal or no SDS concentration.

Results: Decellularisation with 0.25% SDS at 0.15ml/min flow rate resulted in high percentage of cells and DNA content removal. Histology and microscopy revealed a smooth and denuded surface with papillary structures. Water is the best solvent to remove all traces of SDS in the tissue as compared to ethanol.

Conclusion: Porcine oesophagus can be decellularised successfully using SDS, before its removal via washing with distilled water as the best solvent tested. The full removal of SDS is vital before the tissue can be utilised as SDS is a strong anionic detergent that can solubilise the proteins and lipids that form the membrane; all of which might affect the healthy tissues where the decellularised tissue is supposed to be sited on if washing is not thorough. The results concluded with a final scaffold that is cell-free with minimal SDS and has a smooth mucosal surface with a well preserved extracellular matrix; which is perfect for use as a replacement for a surgically removed diseased organ in the treatment of oesophageal cancer.