Scalable Production of Monodisperse Bioactive Spider Silk Nanowires

Abstract

Elongated protein‐based micro‐ and nanostructures are of great interest for a wide range of biomedical applications, where they can serve as a backbone for surface functionalization and as vehicles for drug delivery. Current production methods for protein constructs lack precise control of either shape and dimensions or render structures fixed to substrates. This work demonstrates production of recombinant spider silk nanowires suspended in solution, starting with liquid bridge induced assembly (LBIA) on a substrate, followed by release using ultrasonication, and concentration by centrifugation. The significance of this method lies in that it provides i) reproducability (standard deviation of length <13% and of diameter <38%), ii) scalability of fabrication, iii) compatibility with autoclavation with retained shape and function, iv) retention of bioactivity, and v) easy functionalization both pre‐ and post‐formation. This work demonstrates how altering the function and nanotopography of a surface by nanowire coating supports the attachment and growth of human mesenchymal stem cells (hMSCs). Cell compatibility is further studied through integration of nanowires during aggregate formation of hMSCs and the breast cancer cell line MCF7. The herein‐presented industrial‐compatible process enables silk nanowires for use as functionalizing agents in a variety of cell culture applications and medical research.

A comprehensive review on the biomedical frontiers of nanowire applications

“… Consistently and uniformly functionalizing surfaces in large-scale nanowire production can be difficult, which may hinder the widespread use of these technologies in industry. [ 212 , 213 ]. Entails applying polymer layers to the nanowire surface to shield it from contamination and create a barrier for patterning.…”

Self-Assembly of RGD-Functionalized Recombinant Spider Silk Protein into Microspheres in Physiological Buffer and in the Presence of Hyaluronic Acid

“…There are, however, unsurpassed thresholds in mass-producing natural spider silk since it is challenging to farm and harvest from spiders, which is why recombinant DNA technology has been employed to develop artificial spider silk materials from key parts of the sequence of various silk proteins (spidroins). , Silk-like materials based on recombinant silk proteins successfully deliver the advantages of the natural material with additional properties, such as scalability, , biofunctionalization, − tunability, , low immune response, and improved biocompatibility. , …”