Purpose To optimise a protocol to produce an acellular porcine cornea scaffold and investigate its mechanical integrity and biocompatibility.
Methods Fresh porcine corneas were decellularised with different detergents over a range of concentrations. Morphological and histological examinations were carried out to detect the major structure of the cornea. Completely acellular cornea scaffolds were subjected to uniaxial tensile testing and reseeding assay.
Results Most protocols resulted in the retention of large numbers of whole cells and cell fragments. Only sodium dodecyl sulfate (SDS; 0.5% or 1%) resulted in total decellularisation at 24 h. Histological analysis of the acellular matrix showed that the corneal stromal cells had been completely removed, collagen fibres were arranged in an orderly fashion, and Bowman's layer and Descemet's membrane were both intact after decellularisation. The ultimate tensile strength of acellular matrix treated with 0.5% SDS for 24 h was not affected significantly compared with that of fresh cornea (p>0.05), whereas there was a significant difference between fresh cornea and cornea treated with 1% SDS (p<0.05). Clusters of corneal epithelial cells were observed on the surface of the matrix.
Conclusion Protocols using SDS (0.5% or 1% for 24 h) were successful for cornea decellularisation. Biomechanical analysis and recellularisation showed that treatment with 0.5% SDS for 24 h was optimal.
- tissue engineering
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