Aspartic acid and epidermal growth factor modified decellularized rabbit conjunctiva for conjunctival reconstruction.

Conjunctival reconstruction is an indispensable part of ocular surface regeneration. Decellularized matrix has been considered as an ideal conjunctival substitution for conjunctival reconstruction. In the present study, we report the use of a decellularized rabbit conjunctiva (DRC) for conjunctival reconstruction in the rabbit surgical trauma model. Prepared by the phospholipase A2 decellularized method, the DRC was nearly DNA free while the collagen structure and natural extracellular matrix (ECM) were well preserved. In order to improve the performance of DRC, aspartic acid (Asp) was used as a spacer arm to crosslink epidermal growth factor (EGF) on the DRC to obtain DRC-Asp-EGF. The conjunctival epithelial cells cultured on the DRC-Asp-EGF showed a higher survival rates and a greater potential to differentiate into conjunctival goblet cells (CGCs) than those on the DRC. Finally, three groups were set to evaluate the transplantation effects in the rabbit surgical trauma model for 28 days: DRC-Asp-EGF group, amniotic membrane (AM) group, and ungrafted group. The DRC-Asp-EGF group was completely re-epithelized, and more CGCs were regenerated than the AM group, while no significant improvements were observed in the ungrafted group. Intact collagen structure, angiogenesis, and no scar formation were also observed in the DRC-Asp-EGF group. These results suggest that DRC-Asp-EGF is a feasible and effective transplant for conjunctival reconstruction and ocular surface regeneration.

Crosslinked Decellularized Porcine Pericardium as a Substrate for Conjunctival Reconstruction.

Seeking for suitable conjunctival reconstruction substitutes to overcome the limitations of current substitutes, such as amniotic membrane, is urgent. Decellularized tissues have become a promising strategy for tissue engineering. In this study, we prepared decellularized porcine pericardium (DPP) scaffolds by the phospholipase A2 method and crosslinked them with aspartic acid (Asp) and human endothelial growth factor (hEGF) to enhance biological performance on the DPP, obtaining DPP-Asp-hEGF scaffolds. In vitro DPP showed lower apoptosis, highly desirable, well preservation of extracellular matrix components, and favorable macro-microstructure, which was confirmed by histology, immunofluorescence, electron microscopy, collagen and DNA quantification, and cytotoxicity assay, compared to the native porcine pericardium (NPP). The crosslinked efficacy of the DPP-Asp-hEGF was furtherer verified by in vitro experiments with Fourier transform infrared (FTIR) and X-ray diffraction (XRD). Through animal models of conjunctiva defect model, the DPP-Asp-hEGF revealed a closed, multilayer epithelium with an equal amount of goblet cells and no indication for conjunctival scarring after 28 days, compared to amniotic membrane (AM) groups and sham groups. These results suggested that DPP-Asp-hEGF can offer a good conjunctival reconstructive substitute both in structure and in function.