A dual-layer silk fibroin scaffold for reconstructing the human corneal limbus

作者：Bray Laura J^*; George Karina A; Hutmacher Dietmar W; Chirila Traian V; Harkin Damien G

来源：Biomaterials, 2012, 33(13): 3529-3538.

DOI：10.1016/j.biomaterials.2012.01.045

• Silk fibroin
• Cornea
• Limbus
• Epithelium
• Mesenchymal stromal cells

Membranes prepared from Bombyx mori silk fibroin have shown potential as a substrate for human limbal epithelial (L-EC) and stromal cell cultivation. Here we present fibroin as a dual-layer construct containing both an epithelium and underlying stroma for corneolimbal reconstruction. We have compared the growth and phenotype of L-EC on non-porous versus porous fibroin membranes. Furthermore, we have compared the growth of limbal mesenchymal stromal cells (L-MSC) in either serum-supplemented medium or the MesenCult-XF (R) culture system within fibroin fibrous mats. The co-culture of L-EC and L-MSC in fibroin dual-layer constructs was also examined. L-EC on porous membranes displayed a squamous monolayer; in contrast. L-EC on non-porous fibroin appeared cuboidal and stratified. Both constructs maintained evidence of corneal phenotype (cytokeratin 3/12) and distribution of Delta Np63(+) progenitor cells. L-MSC cultivated within fibroin fibrous mats in serum-supplemented medium contained less than 64% of cells expressing the characteristic MSC phenotype of CD73(+)CD90(+)CD105(+) after two weeks, compared with over 81% in MesenCult-XF (R) medium. Dual-layer fibroin scaffolds consisting of L-EC and L-MSC maintained a similar phenotype as on the separate layers. These results support the feasibility of a 3D engineered limbus constructed from B. mori silk fibroin, and warrant further studies into the potential benefits it offers to corneolimbal tissue regeneration.

• 出版日期2012-5