Abstract
While fibroin isolated from the cocoons of domesticated silkworm Bombyx mori supports growth of human corneal limbal epithelial (HLE) cells, the mechanism of cell attachment remains unclear. In the present study we sought to enhance the attachment of HLE cells to membranes of Bombyx mori silk fibroin (BMSF) through surface functionalization with an arginine-glycine-aspartic acid (RGD)-containing peptide. Moreover, we have examined the response of HLE cells to BMSF when blended with the fibroin produced by a wild silkworm, Antheraea pernyi, which is known to contain RGD sequences within its primary structure. A procedure to isolate A. pernyi silk fibroin (APSF) from the cocoons was established, and blends of the two fibroins were prepared at five different BMSF/APSF ratios. In another experiment, BMSF surface was modified by binding chemically the GRGDSPC peptide using a water-soluble carbodiimide. Primary HLE were grown in the absence of serum on membranes made of BMSF, APSF, and their blends, as well as on RGD-modified BMSF. There was no statistically significant enhancing effect on the cell attachment due to the RGD presence. This suggests that the adhesion through RGD ligands may have a complex mechanism, and the investigated strategies are of limited value unless the factors contributing to this mechanism become better known.
Highlights
Silk proteins have been introduced as biomaterials in the early 1990s through Minoura‘s seminal papers [1,2,3]
The silk fibroin isolated from the cocoons produced by the larvae of domesticated silkmoth Bombyx mori, (BMSF), has been by far the most investigated silk substratum for cells [2,3,5,17,18,19,20,21]
Considering the essential role played in cell adhesion process by the integrin binding sites located on the cell surface, the mechanism of cell attachment to silk fibroins is debatable and worth to investigate
Summary
Silk proteins have been introduced as biomaterials in the early 1990s through Minoura‘s seminal papers [1,2,3]. The two principal constitutive proteins (fibroin and sericin) of the silk threads produced by the larvae of certain silkmoths in class Insecta have been investigated extensively for biomaterials and tissue engineering applications [4,5,6,7,8,9,10,11,12,13,14,15], apparently little or no successful usage in a clinical setting has been reported so far [16]. A typical ligand peptide motif is the arginine-glycine-aspartic acid (RGD) sequence found in fibronectin. When such motifs are present on a substratum, and are sterically exposed, they can promote the cells‘
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