Biochemical, thermal and anatomical characterization of the decellularized corneal scaffold

Abstract

Purpose : Tissue engineering goal is to assemble functional constructs that restore, maintain, or improve damaged tissues. One of the strategies that tissue engineering has been used is the decellularization process of tissues for understand the extracellular matrix as scaffold. In the ophthalmology field concerning the corneal regeneration, this process could be a promising technique. Thus, the aim of this study was to characterize the physicochemical, thermal and morphological properties of decellularized process of pig corneas with SDS detergent.

Methods : Porcine corneal buttons were donated for the study. For start the decellularization process, corneal tissue was washed in PBS with antibiotics for 15 min under stirring at room temperature, subsequently the corneal tissue was subjected to immersion in 0.5% SDS with constant agitation for 24 h. After the period, decellularized corneal were washed with PBS and antibiotics for detergent elimination. Decellularized tissue obtained then was subjected to a physicochemical characterization by FTIR, TGA, DSC and by morphological analysis by SEM.

Results : Morphological characterization of decellularized corneal showed the intact organization of extracellular matrix of the tissue with aligned collagen fibers with sizes around of 471.70 - 254.41 nm. Moreover, the physicochemical characterization by FTIR showed the vibrational peaks of amide II and III of collagen protein and TGA curve showed the thermal and mass properties of corneal scaffold

Conclusions : Characterization of decellularized corneal reveals that the scaffold retains chemical, thermal and anatomical compositional properties similar to those of the human native cornea. The result indicates that extracellular fibrillar matrix is not affected by the process of decellularization, so this scaffold could be an option for future studies about the biocompatibility and repopulation by limbal stem cells.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

 

SEM images of the corneal scaffold with the collagen fiber size (a, b), TGA analysis of the decellularized corneal scaffold (c) and FTIR spectra that showed the vibrational groups of the corneal scaffold (d).