Highly Adhesive Amyloid-Polyphenol Hydrogels for Cell Scaffolding.

Rationally designing microstructures of soft hydrogels for specific biological functionalization is a challenge in tissue engineering applications. A novel and affordable soft hydrogel scaffold is constructed here by incorporating polyphenol modules with lysozyme amyloid fibrils (Lys AFs) via non-covalent self-assembly. Embedded polyphenols not only trigger hydrogel formation but also determine gel behavior by regulating the polyphenol gallol density and complex ratio. The feasibility of using a polyphenol-Lys AF hydrogel as a biocompatible cell scaffold, which is conducive to cell proliferation and spreading, is also shown. Notably, introducing polyphenols imparts the corresponding hydrogels a superior cell bioadhesive efficiency without further biofunctional decoration and thus may be successfully employed in both healthy and cancer cell lines. Confocal laser scanning microscopy also reveals that the highly expressed integrin-mediated focal adhesions form due to stimulation of the polyphenol-AF composite hydrogel, direct cell adhesion, proliferation, and spreading. Overall, this work constitutes a significant step forward in creating highly adhesive tissue culture platforms for in vitro culture of different cell types and may greatly expand prospects for future biomaterial design and development.

Designed modular protein hydrogels for biofabrication.

Designing proteins that fold and assemble over different length scales provides a way to tailor the mechanical properties and biological performance of hydrogels. In this study, we designed modular proteins that self-assemble into fibrillar networks and, as a result, form hydrogel materials with novel properties. We incorporated distinct functionalities by connecting separate self-assembling (A block) and cell-binding (B block) domains into single macromolecules. The number of self-assembling domains affects the rigidity of the fibers and the final storage modulus G' of the materials. The mechanical properties of the hydrogels could be tuned over a broad range (G' = 0.1 - 10 kPa), making them suitable for the cultivation and differentiation of multiple cell types, including cortical neurons and human mesenchymal stem cells. Moreover, we confirmed the bioavailability of cell attachment domains in the hydrogels that can be further tailored for specific cell types or other biological applications. Finally, we demonstrate the versatility of the designed proteins for application in biofabrication as 3D scaffolds that support cell growth and guide their function. STATEMENT OF SIGNIFICANCE: Designed proteins that enable the decoupling of biophysical and biochemical properties within the final material could enable modular biomaterial engineering. In this context, we present a designed modular protein platform that integrates self-assembling domains (A blocks) and cell-binding domains (B blocks) within a single biopolymer. The linking of assembly domains and cell-binding domains this way provided independent tuning of mechanical properties and inclusion of biofunctional domains. We demonstrate the use of this platform for biofabrication, including neural cell culture and 3D printing of scaffolds for mesenchymal stem cell culture and differentiation. Overall, this work highlights how informed design of biopolymer sequences can enable the modular design of protein-based hydrogels with independently tunable biophysical and biochemical properties.

Lactoferrin network with MC3T3-E1 cell proliferation, auxiliary mineralization, antibacterial functions: A multifunctional coating for biofunctionalization of implant surfaces.

Developing biocompatible, low-immunoreactive, and antibacterial implants are challenging yet fundamental to osteosynthesis. In this study, mineralization-stimulative and antibacterial networking nanostructures are assembled via amyloid-like aggregation of lactoferrin (LF) triggered by reducing the intramolecular disulfide bonds. Due to the adhesive property of their rich ß-sheet architecture, the LF networks are amenable to the deposition upon the surface of various implant materials, functionalizing the implants with cell-proliferative, mineralization-stimulative, and antibacterial properties. Specifically, the abundant functional groups and amino acids exposed on the surface of LF networks provide abundant functional microdomains for subsequent mineralization of different forms of calcium ions and promote the formation of hydroxyapatite (HAp) crystals in simulated body fluids. We further demonstrate that the LF network inherits the innate antibacterial properties of LF and exerts a synergistic antibacterial ability with surface-enriched positively charged and hydrophobic amino acid residues, disrupting bacterial biofilm formation, enhancing microbial cell wall perturbation, and ultimately leading to microbial death. The results underscore the feasibility of the LF network as a multifunctional coating on bioscaffold surfaces, which may provide insight into its future applications in next-generation artificial bone implants with bacterial/biofilm clearance and bone tissue remodeling capabilities.

Fiber-Tip Polymer Microcantilever for Fast and Highly Sensitive Hydrogen Measurement.

Hydrogen as an antioxidant gas has been widely used in the medical and biological fields for preventing cancer or treating inflammation. However, controlling the hydrogen concentration is crucial for practical use due to its explosive property when its volume concentration in air reaches the explosive limit (4%). In this work, a polymer-based microcantilever (µ-cantilever) hydrogen sensor located at the end of a fiber tip is proposed to detect the hydrogen concentration in medical and biological applications. The proposed sensor was developed using femtosecond laser-induced two-photon polymerization (TPP) to print the polymer µ-cantilever and magnetron sputtering to coat a palladium (Pd) film on the upper surface of the µ-cantilever. Such a device exhibits a high sensitivity, roughly -2 nm %-1 when the hydrogen concentration rises from 0% to 4.5% (v/v) and a short response time, around 13.5 s at 4% (v/v), making it suitable for medical and environmental applications. In addition to providing an ultracompact optical solution for fast and highly sensitive hydrogen measurement, the polymer µ-cantilever fiber sensor can be used for diverse medical and biological sensing applications by replacing Pd with other functional materials.