As a growing public health concern, the worldwide spread of antimicrobial resistance urges the development of new therapies. Antibacterial photodynamic therapy (a-PDT) may be an alternative to conventional antibiotic therapy. Herein we report the synthesis and characterization of seven original reactive oxygen species (ROS)-producing ruthenium(II) polypyridyl complexes. These are part of a collection of 17 derivatives varying in terms of the nature of the substituent(s), molecular symmetry, electrical charge, and counterions. They were characterized by considering 1) their physical properties (absorption coefficient at irradiation wavelength, O generation quantum yield, luminescence) and 2) their antibacterial activity in a series of photodynamic assays using Gram-positive and Gram-negative bacteria of clinical relevance. The results unveiled some structure-activity relationships: one derivative that combines multiple beneficial features for a-PDT was effective against all the bacteria considered, regardless of their Gram status, species, or antibiotic resistance profile. This systematic study could guide the design of next-generation ruthenium-based complexes for enhanced antibacterial photodynamic strategies.

The synthesis of novel amphiphilic hyaluronic acid (HYA) and poly(lactic acid) (PLA) block copolymers is reported as the key element of a strategy to detect the presence of pathogenic bacterial enzymes. In addition to the formation of defined HYA-block-PLA assemblies, the encapsulation of fluorescent reporter dyes and the selective enzymatic degradation of the capsules by hyaluronidase and proteinase K are studied. The synthesis of the dual enzyme-responsive HYA-b-PLA is carried out by copper-catalyzed Huisgen 1,3-dipolar cycloaddition. The resulting copolymers are assembled in water to form vesicular structures, which are characterized by scanning electron microscopy, transmission electron microscopy, dynamic light scattering (DLS), and fluorescence lifetime imaging microscopy (FLIM). DLS measurements show that both enzymes cause a rapid decrease in the hydrodynamic diameter of the nanocapsules. Fluorescence spectroscopy data confirm the liberation of encapsulated dye, which indicates the disintegration of the capsules and validates the concept of enzymatically triggered payload release. Finally, cytotoxicity assays confirm that the HYA-b-PLA nanocapsules are biocompatible with primary human dermal microvascular endothelial cells.

Stimuli-responsive polymersomes that are selectively cleaved by enzymes of pathogenic bacteria are herein reported. The local disruption of the polyester wall in poly(ethylene glycol)-block-poly(lactic acid) (PEG-b-PLA) polymersomes filled with reporter dyes owing to enzymatic degradation by proteinase K was monitored by fluorescence lifetime imaging microscopy. Capsule opening occurred on timescales of minutes to hours, leading to the release of the dye, followed by a slow capsule disintegration, and finally cleavage into monomeric lactic acid over several weeks. These nanocontainers represent a promising element in novel theranostic systems for potential application in advanced wound dressings.

This study reports on the characterization of reporter dye‐loaded block copolymer vesicles (polymersomes) of PS115‐b‐PAA15 (polystyrene‐block‐poly(acrylic acid)) and PEG114‐b‐PLA167 (poly(ethylene glycol)‐block‐poly(lactic acid)) and their drying behavior by fluorescence lifetime imaging microscopy (FLIM). The characteristic changes of the fluorescence decay components of the dye calcein incorporated in the three different local nanoenvironments, namely, the solvated dye, dye associated with the vesicle wall, and dried agglomerated dye, are observed by FLIM during the drying of vesicles. The amplitude ratio R1/2 of the components attributed to the solvated dye in the vesicle interior with a lifetime τ1 ≈ 3.9 ns to the one attributed to calcein associated with the wall with a lifetime τ2 ≈ 1.5 ns is found to decrease exponentially with drying time. The time constants, which are found to depend linearly on the radius of the vesicle, yield by extrapolation to zero vesicle diameter an estimate of the polymersome wall thickness. For PS115‐b‐PAA15 and PEG114‐b‐PLA167 vesicle wall thicknesses r0 of 16 ± 5 nm and 28 ± 4 nm, respectively, are observed, which are in favorable agreement with transmission electron and atomic force microscopy data.

Enzymatic degradation of polymers has advantages over standard degradation methods, such as soil burial and weathering, which are time-consuming and cannot provide time-resolved observations. We have developed a microfluidic device to study the degradation of single microparticles. The enzymatic degradation of poly (1,4-butylene adipate-co-terephthalate) (PBAT) microparticles was studied using Novozym 51032 cutinase. PBAT microparticles were prepared via an oil-in-water emulsion solvent removal method, and their morphology and chemical composition were characterized. Then, microparticles with varying diameters of 30−60 μm were loaded into the microfluidic chip. Enzyme solutions at different concentrations were introduced to the device, and changes in the size and transparency of PBAT microparticles were observed over time. The physicochemical properties of degraded products were analyzed by FT-IR, NMR, mass spectrometry, and differential scanning calorimetry. The degradation process was also performed in bulk, and the results were compared to those of the microfluidic method. Our analysis confirms that the degradation process in both bulk and microfluidic methods was similar. In both cases, degradation takes place on aliphatic and soft segments of PBAT. Our findings serve as a proof of concept for a microfluidic method for easy and time-resolved degradation analysis, with degradation results comparable to those of conventional bulk methods.