RESUMO
This study investigates the effects of environmentally-relevant concentrations of fluoxetine (FLX, commercial name: Prozac) on wound healing. Pollution of water systems with pharmaceutical and personal care products, including antidepressants such as FLX and other selective serotonin reuptake inhibitors, is a growing environmental concern. Environmentally-relevant FLX concentrations are known to impact physiological functions and behaviour of aquatic animals, however, the effects of exposure on humans are currently unknown. Using a combination of human skin biopsies and a human keratinocyte cell line, we show that exposure to environmental FLX promotes wound closure. We show dose-dependent increases in wound closure with FLX concentrations from 125 ng/l. Using several -omics and pharmaceutical approaches, we demonstrate that the mechanisms underlying enhanced wound closure are increased cell proliferation and serotonin signalling. Transcriptomic analysis revealed 350 differentially expressed genes after exposure. Downregulated genes were enriched in pathways related to mitochondrial function and metabolism, while upregulated genes were associated with cell proliferation and tissue morphogenesis. Kinase profiling showed altered phosphorylation of kinases linked to the MAPK pathway. Consistent with this, phosphoproteomic analyses identified 235 differentially phosphorylated proteins after exposure, with enriched GO terms related to cell cycle, division, and protein biosynthesis. Treatment of skin biopsies and keratinocytes with ketanserin, a serotonin receptor antagonist, reversed the increase in wound closure observed upon exposure. These findings collectively show that exposure to environmental FLX promotes wound healing through modulating serotonin signalling, gene expression and protein phosphorylation, leading to enhanced cell proliferation. Our results justify a transition from the study of behavioural effects of environmental FLX in aquatic animals to the investigation of effects of exposure on wound healing in aquatic and terrestrial animals, including direct impacts on human health.
RESUMO
During the last decades, there has been growing interest in the application of functionalized mesoporous nanomaterials as stimuli-responsive carriers for drug delivery. However, at present there is not a standardized methodology to evaluate their performance. The limitations of the different techniques reported in literature give rise to the necessity for new, simple, and cost-effective alternatives. This work constitutes a step forward in the development of advanced in vitro procedures for testing the behavior of nanocarriers, proposing a novel microfluidic platform. To test the capacity of the reported tool, the performance of amino-functionalized MCM-41 nanoparticles has been assessed. These materials show a pH-responsive mechanism, which prevents the drug release at acidic conditions, maximizing its distribution at neutral pH, thus, the selected release medium mimicked gastrointestinal conditions. As a first approximation, the delivery of Ru(bipy)32+ was evaluated, proving the advantages of the proposed microfluidic system: i) continuous flow of particles and media, ii) rigorous control of the residence time, temperature and pH, iii) enhanced mixing, iv) possibility to simulate different human body conditions and, v) possible integration with the continuous synthesis of nanocarriers. Finally, the microfluidic tool was used to analyze the delivery of the anti-inflammatory drug ibuprofen.