Bio-fabrication of stem-cell-incorporated corneal epithelial and stromal equivalents from silk fibroin and gelatin-based biomaterial for canine corneal regeneration

Abstract

Corneal grafts are the imperative clinical treatment for canine corneal blindness. To serve the growing demand, this study aimed to generate tissue-engineered canine cornea in part of the corneal epithelium and underlying stroma based on canine limbal epithelial stem cells (cLESCs) seeded silk fibroin/gelatin (SF/G) film and canine corneal stromal stem cells (cCSSCs) seeded SF/G scaffold, respectively. Both cell types were successfully isolated by collagenase I. SF/G corneal films and stromal scaffolds served as the prospective substrates for cLESCs and cCSSCs by promoting cell adhesion, cell viability, and cell proliferation. The results revealed the upregulation of tumor protein P63 (P63) and ATP-binding cassette super-family G member 2 (Abcg2) of cLESCs as well as Keratocan (Kera), Lumican (Lum), aldehyde dehydrogenase 3 family member A1 (Aldh3a1) and Aquaporin 1 (Aqp1) of differentiated keratocytes. Moreover, immunohistochemistry illustrated the positive staining of tumor protein P63 (P63), aldehyde dehydrogenase 3 family member A1 (Aldh3a1), lumican (Lum) and collagen I (Col-I), which are considerable for native cornea. This study manifested a feasible platform to construct tissue-engineered canine cornea for functional grafts and positively contributed to the body of knowledge related to canine corneal stem cells.

Agar/gelatin hydro-film containing EGF and Aloe vera for effective wound healing

“…facilitated by the RGD motifs in gelatin, which resembled integrin mediated cell attachment. 16,48,49 It is worth to mention that although AV has shown to enhance cell attachment due to the ability of glucomannan to bind on b2-integrins, we did not observe any differences between G and GA hydro-films 16 Anti-inflammatory study. To determine the anti-inflammatory activity of the hydro-films, we followed the experimental design represented in Fig.…”

Advances in Biomaterials for Corneal Regeneration

“…Studies revealed endogenous ECM production after 14 days, thus mimicking native cornea. [16]. Pitarresi and coworkers were the first ones to use hydrogels based on hyaluronic acid (HA) cross-linked with polyaspartamide derivative (PHEA-EDA) as an alternative to amniotic membrane for the delivery of limbal stem cells (LSCs).…”