Crosslinked Decellularized Porcine Pericardium as a Substrate for Conjunctival Reconstruction

Abstract

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.

Biomaterials and tissue engineering strategies for posterior lamellar eyelid reconstruction: Replacement or regeneration?

“…Most importantly, due to the lack of posterior lamellar cellular components, these grafts barely provide a smooth epithelialized surface and restore secretory function. Although some studies have reported that implanted decellularized matrixes could support cell growth both in vitro 84 and in vivo, 85 concomitant fibrosis rather than complete reepithelization was observed on the surface of the biomaterials upon implantation in animal models 10,109 . Cellular approaches aim to restore the full function of the posterior lamella by implanting a tissue‐engineered tarsal or conjunctival equivalent into the defect area, which can be referred to as “regenerative surgery” and represents a future direction in this field.…”

“…Ideally, bioengineered posterior lamellar substitutes should restore key structures and functions of the eyelid without additional donor‐site morbidities or postoperative complications. To date, many innovative native tissue grafts, 10 , 11 biomaterials, 12 , 13 and bioengineered tissues 14 , 15 have been proposed as posterior lamellar substitutes, some of which have been clinically used for eyelid reconstruction and have yielded promising preliminary results. 11 , 14 , 15 The bioengineering of posterior lamellar substitutes can be categorized into two distinct approaches: (1) acellular approaches, in which natural or synthetic acellular biomaterials are transplanted into defect areas to act as bioscaffolds for guiding tissue regeneration in vivo by leveraging endogenous cells and microenvironments (Figure 4a ); and (2) cellular approaches, in which bioscaffolds are precellularized in vitro to mimic specific functions of the target tissue and then transplanted to promote tissue regeneration (Figure 4b ).…”

Crosslinked modified decellularized rabbit conjunctival stroma for reconstruction of tissue-engineered conjunctiva in vitro

“…E shows that the rate of cells in apoptosis in the DRCS group (9.23 ± 2.72%) is much higher than in the DRCS-Asp-hEGF (0.13 ± 0.06%). Despite these promising reports on the transplantation of acellular scaffolds for defect reconstruction [9,11], there is still scope to conduct recellularization protocols to realize clinical applications. Currently, protocols for recellularization mainly include tissue sectioning, perfusion, and direct injection [28].…”

“…Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were performed, as described previously [11]. In brief, the samples (native rabbit conjunctiva (NRC) and DRCS) were fixed using an electron microscopy fixative for 2 h at room temperature (RT) and then post-fixed with 1% OsO 4 (Ted Pella Inc., USA) for 1 h at RT after washing in PBS (pH 7.4) for 15 min thrice.…”

Human pericardial extracellular matrix: An implantation platform for cardiac tissue engineering

“…In brief, pericardium grafts have been applied to correct intracardiac and diaphragm defects (Ricci et al, 2014;Yaliniz et al, 2014;Ascaso et al, 2021), as well as ischemic ventricular septal defect after acute MI (Mihalj et al, 2022). On the other hand, non-cardiac pericardium-based applications include replacement of brain dura mater (Sun et al, 2018), bleb repair, conjunctival reconstruction, cover of severe corneal wound or tendon elongation in the eye (Niegowski et al, 2020;Ashena et al, 2021;Hedergott et al, 2021;Chen et al, 2022), odontology (Solakoğlu et al, 2022) and eardrum reconstruction (de Dorldodot et al, 2013;Sainsbury et al, 2022). Furthermore it has been employed to generate a variety of bioprostheses such as vascular grafts, patches for abdominal or vaginal wall reconstruction and heart valves (Rémi et al, 2010).…”