In vitro chemical treatment of silk increases the expression of pro-inflammatory factors and facilitates degradation in rats

Abstract

Objectives: Silk fiber is difficult to degrade in vivo, which limits its application in tissue engineering materials such as artificial nerves. Therefore, in this study aim to promote its degradation in vivo by chemical treating silk fibers in vitro. Materials and methods: Sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), scanning electron microscopy (SEM) observations, mechanical test, Fourier transform infrared spectroscopy (FT-IR) measurements were used to investigate the degradation effect of chemicals (hydrochloric acid, phosphoric acid, acetic acid, sodium hydroxide, calcium hydroxide, sodium bicarbonate, and calcium chloride) on silk fiber in vitro. Immunofluorescence staining and transcriptome analysis were used to investigate the effect of inflammatory factors on the degradation of chemically treated silk fiber in rats. Results: (1) Silks were separated into finer fibers in each group. (2) FT-IR absorption peaks of amides I, II, and III overlap in each group. (3) Silk degradation degree in each group was higher than that in an untreated group. The calcium chloride-treated group was completely degraded. (4) Fibronectin, collagen I, collagen III, integrin α and CD68 were immunofluorescence positive in all vegetation section. (5) There were no significant differences in the expressions of collagen I, collagen III, and fibronectin in the vegetations formed on the 14th day of subcutaneous implantation, while integrin α, CD68, TNF-α, IL-1b, and IL-23 express at higher levels with IL-10 at lower levels. Conclusions: All chemicals could completely degrade silk; however, their degradation products were not the same. The chemicals change the mechanical properties of silk by separating it into finer fibers, which increase the contact surface area between the silk and tissue fluid, accelerating the degradation of monofilaments in vivo by promoting inflammation and macrophage activity through the increased and decreased expressions of pro- and anti-inflammatory factors, respectively.