Genetically engineered bi-functional silk material with improved cell proliferation and anti-inflammatory activity for medical application

Abstract

Functional silk is a promising material for future medical applications. These include fabrication of diverse silk fiber and silk protein-regenerated biomaterials such as silk sutures, hydrogel, films, and 3D scaffolds for wound healing and tissue regeneration and reconstruction. Here, a novel bi-functional silk with improved cell proliferation and anti-inflammatory activities was created by co-expressing the human basic fibroblast growth factor (FGF2) and transforming growth factor-β1 (TGF_β1) genes in silkworm. First, both FGF2 and TGF_β1 genes were confirmed to be successfully expressed in silk thread. The characterization of silk properties by SEM, FTIR, and mechanical tests showed that this new silk (FT silk) had a similar diameter, inner molecular composition, and mechanical properties as those of normal silk. Additionally, expressed FGF2 and TGF_β1 proteins were continuously and slowly released from FT silk for one week. Most importantly, the FGF2 and TGF_β1 contained in FT silk not only promoted cell proliferation by activating the ERK pathway but also significantly reduced LPS-induced inflammation responses in macrophages by mediating the Smad pathway. Moreover, this FT silk had no apparent toxicity for cell growth and caused no cell inflammation. These properties suggest that it has a potential for medical applications. STATEMENT OF SIGNIFICANCE: Silk spun by domestic silkworm is a promising material for fabricating various silk protein regenerated biomaterials in medical area, since it owes good biocompatibility, biodegradability and low immunogenicity. Recently, fabricating various functional silk fibers and regenerated silk protein biomaterials which has ability of releasing functional protein factor is the hot point field. This study is a first time to create a novel bi-functional silk material with the improved cell proliferation and anti-inflammatory activity by genetic engineered technology. This novel silk has a great application potential as new and novel medical material, and this study also provides a new strategy to create various functional or multifunctional silk fiber materials in future.

Keywords: Binary-functional silk; Genetic engineering; Human basic fibroblast growth factor; Human transforming growth factor-β1; Silkworm.