Dextran preserves native corneal structure during decellularization

Corneal decellularization has become an increasingly popular technique for generating scaffolds for corneal regeneration. Most decellularization procedures result in tissue swelling, thus limiting their application. Here, the use of a polysaccharide, dextran, to reduce swelling and conserve the native corneal structure during decellularization was investigated. Corneas were treated with 1% Triton X-100, 0.5% sodium dodecyl sulfate, and nucleases under constant rotation followed by extensive washing. To reduce swelling, decellularization solutions were supplemented with 5% dextran either throughout the whole decellularization process or during the washing cycles only. Quantitative analysis of DNA content showed a 96% reduction after decellularization regardless of the addition of dextran. Dextran resulted in a significant reduction in swelling from 3.85 ± 0.43 nm without to 1.94 ± 0.29-2.01 ± 0.37 nm (p 0.05. Dextran can counteract the detrimental effects of decellularizing agents on the biomechanical properties of the tissue resulting in similar compressive moduli (mean before decellularization: 5.40 ± 1.18 kPa; mean after decellularization with dextran: 5.64 ± 1.34 kPa, p > 0.05). Cells remained viable in the presence of decellularized scaffolds. The findings of this study indicate that dextran not only prevents significant corneal swelling during decellularization but also enhances the maintenance of the native corneal ultrastructure

Characterizing the role of dextran in the decellularization of porcine corneas [Abstract]

Characterizing the role of dextran in the decellularization of porcine corneas [Abstract

Ultrastructural maintenance of decellularized corneas using dextran [abstract]

PURPOSE. Corneal decellularization has emerged as a promising alternative to traditional tissue-engineering strategies for the creation of corneal replacements for transplantation. However, decellularization methods can lead to swelling of the cornea, limiting its potential use as a scaffold. In this study, we propose the use of a complex polysaccharide, dextran, to reduce this swelling and maintain the native dimensions and architecture of the cornea. METHODS. Porcine corneal buttons were treated with Triton X-100, SDS and nucleases under constant rotation followed by a washing step. To prevent corneal swelling, the decellularization solution was supplemented with dextran. This solution was added to one group throughout the decellularization process and to a second group during the washing cycle. The resulting acellular scaffolds were systematically evaluated by histological and biochemical analyses, in addition, the ultrastructure of the cornea was examined by transmission electron microscopy (TEM). RESULTS. Results demonstrated that the combination of detergents and nucleases effectively removed the majority of cellular material from the cornea. Furthermore, the addition of dextran prevented significant swelling when used throughout the protocol or only during the washing process. After soaking in glycerol a degree of transparency was returned to all decellularized corneas suggesting maintenance of the extracellular matrix. However, TEM analysis confirmed that dextran must be present throughout the decellularization process to preserve the native ultrastructure of the cornea. CONCLUSIONS. The findings of this study indicate that the addition of dextran to the decellularization process not only prevents significant corneal swelling but also enhances the maintenance of the native ultrastructure of the cornea