We report on the fabrication of efficient antibacterial substrates selective for bacteria, i.e., noncytotoxic against mammalian cells. The strategy proposed is based on the different size of bacteria (1-4 μm) in comparison with mammalian cells (above 20 μm) that permit the bacteria to enter in contact with the inner part of micrometer-sized pores where the antimicrobial functionality are placed. On the contrary, mammalian cells, larger in terms of size, remain at the top surface, thus reducing adverse cytotoxic effects and improving the biocompatibility of the substrates. For this purpose, we fabricated well-ordered functional microporous substrates (3-5 μm) using the breath figures approach that enabled the selective functionalization of the pore cavity, whereas the rest of the surface remained unaffected. Microporous surfaces were prepared from polymer blends comprising a homopolymer (i.e., polystyrene) and a block copolymer (either polystyrene-b-poly(dimethylaminoethyl methacrylate) (PDMAEMA) or a quaternized polystyrene-b-poly(dimethylaminoethyl methacrylate)). As a result, porous surfaces with a narrow size distribution and a clear enrichment of the PDMAEMA or the quaternized PDMAEMA block inside the pores were obtained that, in the case of the quaternized PDMAEMA, provided an excellent antimicrobial activity to the films.
We describe herein a novel strategy for the fabrication of efficient 3D printed antibacterial scaffolds. For this purpose, both the surface topography as well as the chemical composition of 3D scaffolds fabricated by additive manufacturing were modified. The scaffolds were fabricated by fused deposition modeling (FDM) using high-impact polystyrene (HIPS) filaments. The surface of the objects was then topographically modified providing materials with porous surfaces by means of the Breath Figures approach. The strategy involves the immersion of the scaffold in a polymer solution during a precise period of time. This approach permitted the modification of the pore size varying the immersion time as well as the solution concentration. Moreover, by using polymer blend solutions of polystyrene and polystyrene-b-poly(acrylic acid) (PS-b-PAA) and a quaternized polystyrene-b-poly(dimethylaminoethyl methacrylate) (PS-b-PDMAEMAQ), the scaffolds were simultaneously chemically modified. The surfaces were characterized by scanning electron microscopy and infrared spectroscopy. Finally, the biological response toward bacteria was explored. Porous surfaces prepared using quaternized PDMAEMA as well as those prepared using PAA confer antimicrobial activity to the films, i.e., were able to kill on contact Staphylococcus aureus employed as model bacteria.
We designed and fabricated highly efficient and selective antibacterial substrates, i.e. surface non-cytotoxic against mammalian cells but exhibiting strong antibacterial activity. For that purpose, microporous substrates (pore sizes in the range of 3-5 m) were fabricated using the breath figures approach (BFs). These substrates have additionally a defined chemical composition in the pore cavity (herein either a poly(acrylic acid) or the antimicrobial peptide Nisin) while the composition of the rest of the surface is identical to the polymer matrix. As a result, considering the differences in size of bacteria (1-4 m) in comparison to mammalian cells (above 10 µm) the bacteria were able to enter in contact with the inner part of the pores where the antimicrobial functionality has been placed. On the opposite, mammalian cells remain in contact with the top surface thus preventing cytotoxic effects and enhancing the biocompatibility of the substrates. The resulting antimicrobial surfaces were exposed to Staphylococcus aureus as a model bacteria and murine endothelial C166-GFP cells. Superior antibacterial performance while maintaining an excellent biocompatibility was obtained by those surfaces prepared using PAA while no evidence of significant antibacterial activity was observed at those surfaces prepared using Nisin.
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