Effect of the chemical modification of the arginyl residue in Bombyx mori silk fibroin on the attachment and growth of fibroblast cells.

Abstract

We prepared matrices of Bombyx mori silk fibroin (SF) with different degrees of modification of arginyl residues by reaction between 1,2-cyclohexanedione (CHD) and SF. Two kinds of SF, namely native SF (NSF), obtained from the silk gland of silkworm larvae, and regenerated SF (RSF), prepared from cocoons of the same silkworm, were used in this study because their amino acid compositions were slightly different from each other. The attachment and growth of mouse fibroblast (L-929) cells on the matrices of the NSF and RSF, in which half or almost all of the arginyl residues were modified (NSF50, RSF50, NSF100, and RSF100), were studied using a cell culture method. Both NSF50 and NSF100 exhibited higher cell attachment than did the unmodified NSF. While the cell growth on NSF50 was not significantly different from that on NSF and NSF100, the growth on NSF100 was higher than that on NSF. The cells attached to NSF50 and NSF100 were extensively spread out and their filopodia were visible by SEM. The cell attachment and growth on RSF were comparable to those on NSF100. Although RSF50 exhibited almost the same cell attachment as did the unmodified RSF, RSF100 exhibited a lower cell attachment than did the unmodified RSF and RSF50. There were no significant differences in the cell growth among RSF series. The cells attached to RSF50 and RSF100 aggregated to form masses, and their filopodia could not be found. The relationship of cell attachment to the basicity of the substrate is considered because the modification of the positively charged arginyl residue changed the basicity of the substrate and the cell attachment on the substrate.