Femtosecond laser-patterned nanopore arrays for surface-mediated peptide treatment

The major goal of this study was to create easy-to-use, reusable substrates capable of storing any peptides or bioactive molecules for a desired period of time until cells uptake them without the need for bioactive molecule or peptide-specific techniques. Nanopore arrays of uniform size and distribution were machined into fused silica substrates using femtosecond laser ablation and loaded with peptides by simple adsorption. The nanopore substrates were validated by examining the effect of N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) loaded nanopores on macrophage phagocytosis and intracellular production of reactive oxygen species (ROS) with and without the pro-inflammatory lipopolysaccharide (LPS). Our results demonstrated that nanopores were generated in a uniform array fashion. Ac-SDKP peptides were stably stored in nanopores and internalized by macrophages. Significant reductions in ROS production and phagocytosis in macrophages were observed over control substrates, even in combination with LPS stimulation, indicating that loading Ac-SDKP peptides in pores significantly improved the anti-inflammatory effects.From the Clinical EditorThis team of scientists intended to create easy-to-use, reusable substrates for storing peptides or bioactive molecules for a desired period of time before cellular uptake occurs, and without the need for bioactive molecule or peptide-specific techniques. They demonstrate the successful generation of nanopores in a uniform array that stably stores Ac-SDKP peptides in the nanopores. When peptides were internalized by macrophages, significant reductions in ROS production and phagocytosis were observed, indicating improved anti-inflammatory effects.

Graphical Abstract(A) SEM of 1 million nanopore array showing uniformity of pore size, shape, and distribution. (B) SEM of a single nanopore showing pore morphology. (C) Z-stack confocal micrograph of empty nanopore array. (D) Z section of FITC-SDKP loaded nanopore array after two days of incubation in culture media. (E) FITC-SDKP visualized in a nanopore array after two days of incubation in culture media. (F) Fluorescent image of Raw 264.7 cells cultured on FITC-SDKP loaded nanopore array, and (G) merged fluorescent and phase contrast image of Raw 264.7 cells cultured on FITC-SDKP loaded nanopore array showing cellular uptake of peptide.Figure optionsDownload high-quality image (203 K)Download as PowerPoint slide