Hierarchical rose-petal surfaces delay the early-stage bacterial biofilm growth

Abstract

A variety of natural surfaces exhibit antibacterial properties; as a result significant efforts in the past decade have been dedicated towards fabrication of biomimetic surfaces that can help control biofilm growth. Examples of such surfaces include rose petals, which possess hierarchical structures like the micro-papillae measuring tens of microns and nano-folds that range in the size of 700 ±100 nm. We duplicated the natural structures on rose-petal surfaces via a simple UV-curable nanocasting technique, and tested the efficacy of these artificial surfaces in preventing biofilm growth using clinically relevant bacteria strains. The rose-petal structured surfaces exhibited hydrophobicity (contact angle~130.8º ±4.3º) and high contact angle hysteresis (~91.0° ±4.9°). Water droplets on rose-petal replicas evaporated following the constant contact line mode, indicating the likely coexistence of both Cassie and Wenzel states (Cassie-Baxter impregnating wetting state). Fluorescent microscopy and image analysis revealed the significantly lower attachment of Staphylococcus epidermidis (86.1± 6.2% less) and Pseudomonas aeruginosa (85.9 ±3.2% less) on the rose-petal structured surfaces, compared with flat surfaces over a period of 2 hours. Extensive biofilm matrix was observed in biofilms formed by both species on flat surfaces after prolonged growth (several days), but was less apparent on rose-petal biomimetic surfaces. In addition, the biomass of S. epidermidis (63.2 ±9.4% less) and P. aeruginosa (76.0 ±10.0% less) biofilms were significantly reduced on the rose-petal structured surfaces, in comparison to the flat surfaces. By comparing P. aeruginosa growth on representative unitary nano-pillars, we demonstrated that hierarchical structures are more effective in delaying biofilm growth. The mechanisms are two-fold: 1) the nano-folds across the hemispherical micro-papillae restrict initial attachment of bacterial cells and delay the direct contacts of cells via cell alignment, and 2) the hemispherical micro-papillae arrays isolate bacterial clusters and inhibit the formation of a fibrous network. The hierarchical features on rose petal surfaces may be useful for developing strategies to control biofilm formation in medical and industrial contexts.