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Decellularisation and characterisation of porcine pleura for lung tissue engineering

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


Trisha Vikranth

Nicholas R Forsyth


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Peer Reviewed Not Peer Reviewed