Decellularisation and characterisation of porcine pleura for lung tissue engineering

Vikranth, Trisha; Dale, Tina; Forsyth, Nicholas R

Abstract

Decellularisation offers a broad range of biomimetic scaffolds of allogeneic and xenogeneic origins, exhibiting innate tissue-specific characteristics. We explored a physico-chemical method for decellularising porcine pleural membranes (PPM) as potential tissue-engineered surrogates for lung tissue repair. Decellularised PPM (dPPM) was characterised with histology, quantitative assays, mechanical testing, and sterility evaluation. Cytotoxicity and recellularisation assays assessed the biocompatibility of dPPM. Haematoxylin and Eosin staining showed an evident reduction in nuclei in dPPM, confirmed with nuclear staining and analysis (****p < 0.0001). Sulphated glycosaminoglycans (sGAG) and collagen histology demonstrated minimal disruption to the structural assembly of the core extracellular matrix (ECM) in dPPM. Confocal imaging demonstrated realignment of ECM fibres in dPPM against native control. Quantitative analysis defined a significant change in the angular distribution (****p < 0.0001) and coherence of fibre orientations (***p < 0.001) in dPPM versus native ECM. DNA quantification indicated ≥ 85% reduction in native nuclear dsDNA in dPPM (**p < 0.001). Collagen and sGAG quantification indicated reductions of both (**p < 0.001). dPPM displayed increased membrane thickness (*p < 0.05). However, the Youngs modulus (447.8 ± 41.9 kPa) and ultimate tensile strength (5080 ± 2034.5 kPa) of dPPM were comparable with that of native controls at (411.3 ± 8.1 kPa) and (3933.3 ± 1734.5), respectively. In vitro cytotoxicity and scaffold biocompatibility assays demonstrated robust human mesothelial cell line (MeT-5A) attachment and viability. Here, we define a decellularisation protocol for porcine pleura that represents a step forward in their potential application as bioscaffolds in lung tissue engineering.

• Journal Article Type: Article
• Acceptance Date: Jun 23, 2023
• Online Publication Date: Jun 23, 2023
• Deposit Date: Oct 31, 2023
• Journal: bioRxiv
• Print ISSN: 2692-8205
• Publisher: Cold Spring Harbor Laboratory Press
• Peer Reviewed: Not Peer Reviewed
• DOI: https://doi.org/10.1101/2023.06.20.545830