Antimicrobial peptides are promising alternatives to traditional antibiotics. A group of selfassembling lipopeptides was formed by attaching an acyl chain to the N-terminus of α-helix forming peptides with the sequence C x -G(IIKK) y I-NH 2 (C x G y , x = 4-12 and y = 2). C x G y selfassemble into nanofibers above their critical aggregation concentrations (CACs). With increasing x, the CACs decrease and the hydrophobic interactions increase, promoting secondary structure transitions within the nanofibers. Antimicrobial activity, determined by the minimum inhibition concentration (MIC), also decreases with increasing x, but the MICs are significantly smaller than the CACs, suggesting effective bacterial membrane disrupting power. Unlike conventional antibiotics, both C 8 G 2 and C 12 G 2 can kill S. aureus and E. coli after only minutes of exposure. C 12 G 2 nanofibers have considerably faster killing dynamics and lower cytotoxicity than their non-aggregated monomers. Antimicrobial activity of peptide aggregates has to date been underexploited and it is found to be a very promising mechanism for peptide design. Detailed evidence for the molecular mechanisms involved are provided, based on super-resolution fluorescence microscopy, ss-NMR, AFM, neutron scattering/reflectivity, CD and Brewster angle microscopy.

Complex and hierarchically functionalized scaffolds composed of micro-and nanoscale structures are a key goal in tissue engineering. The combination of three-dimensional (3D) printing and electrospinning enables the fabrication of these multiscale structures. This study presents a polycaprolactone 3D-printed and electrospun scaffold with multiple mesh layers and fiber densities. The results show successful fabrication of a dual-scale scaffold with the 3D-printed scaffold acting as a gap collector with the printed microfibers as the electrodes and the pores a series of insulating gaps resulting in aligned nanofibers. The electrospun fibers are highly aligned perpendicular to the direction of the printed fiber and form aligned meshes within the pores of the scaffold. Mechanical testing showed no significant difference between the number of mesh layers whereas the hydrophobicity of the scaffold increased with increasing fiber density. Biological results indicate that increasing the number of mesh layers improves cell proliferation, migration, and adhesion. The aligned nanofibers within the microscale pores allowed enhanced cell bridging and cell alignment that was not observed in the 3D-printed only scaffold. These results demonstrate a facile method of incorporating low-density and aligned fibers within a 3D-printed scaffold that is a promising development in multiscale hierarchical scaffolds where alignment of cells can be desirable.

Particle tracking microrheology was used to investigate the viscoelasticity of Staphylococcus aureus biofilms grown in microfluidic cells at various flow rates and when subjected to biofilm-degrading enzymes. Biofilm viscoelasticity was found to harden as a function of shear rate but soften with increasing height away from the attachment surface in good agreement with previous bulk results. Ripley's K-function was used to quantify the spatial distribution of the bacteria within the biofilm. For all conditions, biofilms would cluster as a function of height during growth. The effects of proteinase K and DNase-1 on the viscoelasticity of biofilms were also investigated. Proteinase K caused an order of magnitude change in the compliances, softening the biofilms. However, DNase-1 was found to have no significant effects over the first 6 h of development, indicating that DNA is less important in biofilm maintenance during the initial stages of growth. Our results demonstrate that during the preliminary stages of Staphylococcus aureus biofilm development, column-like structures with a vertical gradient of viscoelasticity are established and modulated by the hydrodynamic shear caused by fluid flow in the surrounding environment. An understanding of these mechanical properties will provide more accurate insights for removal strategies of early-stage biofilms.

Sensors that harness quantum mechanical effects can enable high sensitivity and high spatial resolution probing of their environment. The nitrogen-vacancy defect in diamond, a single, optically accessible electronic spin, is a promising quantum sensor that can operate in soft and living systems and provides nanoscale spatial resolution when hosted inside a diamond nanoparticle. Nanodiamond quantum sensors are nontoxic, amenable to surface functionalization, and can be introduced into a variety of living systems. The optical readout of the spin provides detailed information about the local electromagnetic and thermal environment in a noninvasive way. In this Perspective, we introduce the different modalities that nanodiamond quantum sensors offer, highlight recent progress in quantum sensing of biological systems, and discuss remaining challenges and directions for future efforts.