Papers by Daniela Uccelletti
International Journal of Molecular Sciences
Nanomaterials are gaining increasing attention as innovative materials in medicine. Among nanomat... more Nanomaterials are gaining increasing attention as innovative materials in medicine. Among nanomaterials, zinc oxide (ZnO) nanostructures are particularly appealing because of their opto-electrical, antimicrobial, and photochemical properties. Although ZnO is recognized as a safe material and the Zn ion (Zn2+) concentration is strictly regulated at a cellular and systemic level, different studies have demonstrated cellular toxicity of ZnO nanoparticles (ZnO-NPs) and ZnO nanorods (ZnO-NRs). Recently, ZnO-NP toxicity has been shown to depend on the intracellular accumulation of ROS, activation of autophagy and mitophagy, as well as stabilization and accumulation of hypoxia-inducible factor-1α (HIF-1α) protein. However, if the same pathway is also activated by ZnO-NRs and how non-cancer cells respond to ZnO-NR treatment, are still unknown. To answer to these questions, we treated epithelial HaCaT and breast cancer MCF-7 cells with different ZnO-NR concentrations. Our results showed that...
Journal of Biomedical Materials Research Part A, 2010
The goal of this study was to generate porous scaffolds from the genetically engineered protein, ... more The goal of this study was to generate porous scaffolds from the genetically engineered protein, an analogue of Nephila clavipes spidroin 1 (rS1/9) and to assess the properties of new rS1/9 scaffolds essential for bioengineering. The salt leaching technique was used to make the rS1/9 scaffolds of interconnected macroporous structure with spontaneously formed micropores. The tensile strength of scaffolds was 18 6 5 N/cm 2. Scaffolds were relatively stable in a phosphate buffer but degraded in oxidizing environment after 11 weeks of incubation. Applicability of the recombinant spidroin 1 as a substrate for cell culture was demonstrated by successful 3T3 cells growth on the surface of rS1/9 films (270 6 20 cells/mm 2 vs. 97 6 8 cells/mm 2 on the glass surface, p < 0.01). The 3T3 fibroblasts readily proliferated within the rS1/9 scaffold (from initially plated 19 6 2 cells/mm 3 to 3800 6 304 cells/mm 3 after 2 weeks). By this time, cells were uniformly distributed between the surface and deeper layers (27% 6 8% and 33% 6 4%, respectively; p > 0.05), whereas the initial distribution was 58% 6 7% and 11% 6 8%, respectively; p < 0.05). The rS1/9 scaffolds implanted subcutaneously into Balb/c mice were well tolerated. Over a 2-month period, the scaffolds promoted an ingrowth of de novo formed vascularized connective tissue elements and nerve fibers. Thus, scaffolds made of the novel recombinant spidroin 1 analogue are potentially applicable in tissue engineering. V
Journal of Nanoparticle Research, 2016
The exploitation of nanomaterials with antimicrobial properties has attracted an ever-growing int... more The exploitation of nanomaterials with antimicrobial properties has attracted an ever-growing interest in the recent years. Carbon-based materials, such as graphene and graphene family materials (GFMs), have gained most of the attention for application in many biomedical fields. Here, we describe the antimicrobial activity of graphene-like (GL) layers derived from the chemical demolition of carbon black, against the planktonic growth of Staphylococcus aureus cells, primary cause of hospital and community-acquired infections, often leading to bacteremia and sepsis. The inhibitory capabilities of GL layers on the formation of S. aureus biofilm are also assessed. The antimicrobial properties seem based mainly on the interaction between GL layers and bacteria surfaces. FESEM and AFM analyses suggest that the GL layers coat the cells as soon as they get in contact with them, as also indicated by the wettability of the GLs.
Frontiers in Molecular Biosciences
High levels of 2-hydroxyisobutyric acid (2-HIBA) were found in urines of patients with obesity an... more High levels of 2-hydroxyisobutyric acid (2-HIBA) were found in urines of patients with obesity and hepatic steatosis, suggesting a potential involvement of this metabolite in clinical conditions. The gut microbial origin of 2-HIBA was hypothesized, however its actual origin and role in biological processes are still not clear. We investigated how treatment with 2-HIBA affected the physiology of the model organism Caenorhabditis elegans, in both standard and high-glucose diet (HGD) growth conditions, by targeted transcriptomic and metabolomic analyses, Coherent Anti-Stokes Raman Scattering (CARS) and two-photon fluorescence microscopy. In standard conditions, 2-HIBA resulted particularly effective to extend the lifespan, delay ageing processes and stimulate the oxidative stress resistance in wild type nematodes through the activation of insulin/IGF-1 signaling (IIS) and p38 MAPK pathways and, consequently, through a reduction of ROS levels. Moreover, variations of lipid accumulation ...
BACKGROUND: Chemical disinfection of surfaces and instruments for infection control in dental uni... more BACKGROUND: Chemical disinfection of surfaces and instruments for infection control in dental units is a relevant practice, especially at the time of antibiotic resistance microbes. Resistant bacteria are in fact, involved in the high incidence of healthcare-acquired infections, recognized as critical emergence in hospitals and clinics around the world. Infected patients disseminate and release many multidrug-resistant Gram-negative and Gram–positive species to other ones and to healthy people: such bacteria share the ability to survive on various hospital surfaces for long periods and for this reason they are difficult to eradicate by cleaning and chemical disinfection. In this regard, the use of nanomaterials as novel and non-traditional antibacterial agents offers new insight in the field of disinfection. Here we report the development of carbon based nanomaterials, such as graphene nanoplatelets (GNPs) decorated with Zinc oxide nanorods (ZNGs) to be used as disinfectant agents i...
Biology, 2021
Calcium silicate-based cements have reached excellent levels of performance in endodontics, provi... more Calcium silicate-based cements have reached excellent levels of performance in endodontics, providing predictable and successful results. To better assess the properties of these bioactive materials, the present study aimed to compare the biocompatibility and antibiofilm properties of ProRoot MTA and Biodentine. Human osteogenic sarcoma (Saos-2) cells were cultured on ProRoot MTA and Biodentine samples or in the presence of both cement extracts. Cell viability assay, measurement of reactive oxygen species (ROS), immunofluorescence analysis, as well as morphological evaluations were conducted. Moreover, Streptococcus mutans was used to assess the biofilm forming ability on ProRoot MTA and Biodentine disks. Finally, both cements were applied in vivo to treat immature permanent teeth affected by reversible pulpitis. Results: Cell viability assay demonstrated that Saos-2 cells had a dose- and time-dependent cytotoxicity to both analyzed cements, although cells exposed to ProRoot MTA sho...
: The Caenorhabditis elegans bus mutants (bacterial unswollen) were isolated by their altered res... more : The Caenorhabditis elegans bus mutants (bacterial unswollen) were isolated by their altered response to the nematode pathogen Microbacterium nematophilum. The bus-2, bus-4 and bus-17 mutants are resistant to infection by this bacterium and to infection by human pathogens Yersinia pestis and Yersinia pseudotuberculosis. Here we extend that list to Staphylococcus aureus. The bus-2, bus-4 and bus-17 mutants each harbors a defect in a different glycosyltransferase involved in O-glycosylation. Our previous glycomics analysis of the reference strains revealed significant O-glycosylation defects. To further investigate the nature of bus mutant pathogen resistance, we examined gene expression for all three mutants and identified four distinct areas of dysregulation: 1) N- and O-glycosylation; 2) innate immune response; 3) protein folding and editing control; and 4) cuticle structure. Increased gene expression of innate immune system components previously seen in response to pathogenic Gra...
European Cells & Materials, 2013
Biosorption represents a competitive technological alternative for heavy metal removal. This tech... more Biosorption represents a competitive technological alternative for heavy metal removal. This technique exploits the property of certain bio-molecules (or types of biomass) to bind and concentrate selected ions or other molecules from aqueous solutions1. The main advantage of biosorption, compared to traditional sorption techniques, is the application of an inexpensive adsorbing material. The scarce mechanical strength and the difficulties of separation from liquid make however, impractical the direct large scale application of non-living biomasses. Immobilization of the biomass within mechanically stable matrices is needed in industrial practice. The choice of the immobilization matrix is of fundamental importance as it determines not only the mechanical stability but also the possibility of the immobilized biomass to participate in the heavy metal removal. In this contribution, the biosorption of copper onto a non-living Saccharomyces cerevisiae wild strain biomass immobilized in calcium alginate matrices is investigated
Frontiers in Materials, 2020
Microorganisms often cause significant damage on historical objects. The archive or library mater... more Microorganisms often cause significant damage on historical objects. The archive or library materials as well as textile or leather artifacts suffer serious attacks that need appropriate care treatments. Several biocide processes have been implemented but often their application does not preserve the material of the good. The objective of this work is the disinfection through ionizing radiation of leather wallpaper from the museum building Palazzo Chigi in Ariccia (Rome, Italy). The controlled sterilization treatments were carried out using X-ray beams to eliminate the microorganisms present on the leather and maintaining unchanged the properties of the constituent material. Some fragments of decorated leather wallpaper, dating back to the 1700s, were irradiated with X-rays up to 5,000 Gy. The amount of microorganisms was evaluated by microbiological analysis before and after X-ray irradiation treatments to identify the dose that inhibits the bacterial load. It will be shown how the results obtained by the application of different chemical-physical techniques (Scanning Electron Microscopy, Fourier Transform Infrared spectroscopy and Light Transmission Analysis) have helped in the evaluation of the impact of the X-rays on leather chemical and physical integrity.
Journal of Nanobiotechnology, 2019
Background: Dental caries is a recognized worldwide public health problem. Despite being one of t... more Background: Dental caries is a recognized worldwide public health problem. Despite being one of the most effective strategies against dental caries, the excessive use of fluorine may result in a potential risk of developing dental fluorosis especially in children under age of six. The purpose of this work is to analyze a fluorine-free toothpaste containing Biomimetic Hydroxyapatite to assess enamel re-mineralizing and repairing properties. Results: The study was performed in vitro and in vivo, comparing the hydroxyapatite toothpaste with two others toothpaste containing different fluorine concentrations. The coating effect of the micro-structured Hydroxyapatite nanoparticles reintegrates the enamel with a biomimetic film reproducing the structure and the morphology of the biologic Hydroxyapatite of the enamel. As demonstrated, the coating is due to the deposit of a new layer of apatite, which presents fewer particles than the natural enamel, not based on the chemical-physical changes occurring in fluorinated toothpastes. Moreover, it shows resistance to brushing as a consequence of chemical bonds between the synthetic and natural crystals of the enamel. Conclusions: The use of Biomimetic Hydroxyapatite toothpastes has proven to be a valuable prevention measure against dental caries in primary dentition since it prevents the risk of fluorosis.
BioMed Research International, 2015
The nematodeCaenorhabditis elegansis widely used as a model system for research on aging, develop... more The nematodeCaenorhabditis elegansis widely used as a model system for research on aging, development, and host-pathogen interactions. Little is currently known about the mechanisms underlying the effects exerted by foodborne microbes. We took advantage ofC. elegansto evaluate the impact of foodborne microbiota on well characterized physiological features of the worms. Foodborne lactic acid bacteria (LAB) consortium was used to feed nematodes and its composition was evaluated by 16S rDNA analysis and strain typing before and after colonization of the nematode gut.Lactobacillus delbrueckii, L. fermentum, andLeuconostoc lactiswere identified as the main species and shown to display different worm gut colonization capacities. LAB supplementation appeared to decrease nematode lifespan compared to the animals fed with the conventionalEscherichia colinutrient source or a probiotic bacterial strain. Reduced brood size was also observed in microbiota-fed nematodes. Moreover, massive accumul...
Applied Microbiology and Biotechnology, 2020
The probiotic bacteria are helpful for nutritional and therapeutic purposes, and they are commerc... more The probiotic bacteria are helpful for nutritional and therapeutic purposes, and they are commercially available in various forms, such as capsules or powders. Increasing pieces of evidence indicate that different growth conditions and variability in manufacturing processes can determine the properties of probiotic products. In recent years, the lipidomic approach has become a useful tool to evaluate the impact that probiotics induce in host physiology. In this work, two probiotic formulations with identical species composition, produced in two different sites, the USA and Italy, were utilized to feed Caenorhabditis elegans, strains and alterations in lipid composition in the host and bacteria were investigated. Indeed, the multicellular organism C. elegans is considered a simple model to study the in vivo effects of probiotics. Nematodes fat metabolism was assessed by gene expression analysis and by mass spectrometry-based lipidomics. Lipid droplet analysis revealed a high accumulation of lipid droplets in worms fed US-made products, correlating with an increased expression of genes involved in the fatty acid synthesis. We also evaluated the lifespan of worms defective in genes involved in the insulin/IGF-1-mediated pathway and monitored the nuclear translocation of DAF-16. These data demonstrated the involvement of the signaling in C. elegans responses to the two diets. Lipidomics analysis of the two formulations was also conducted, and the results indicated differences in phosphatidylglycerol (PG) and phosphatidylcholine (PC) contents that, in turn, could influence nematode host physiology. Results demonstrated that different manufacturing processes could influence probiotics and host properties in terms of lipid composition. Key points • Probiotic formulations impact on Caenorhabditis elegans lipid metabolism; • Lipidomic analysis highlighted phospholipid abundance in the two products; • Phosphocholines and phosphatidylglycerols were analyzed in worms fed the two probiotic formulations.
The control of bacterial growth is one of the most challenging environmental issues, since surfac... more The control of bacterial growth is one of the most challenging environmental issues, since surfaces exposed to media rich in microorganisms can be damaged or lose their functionality. In this work, we have investigated the antimicrobial properties of Graphene Nanoplatelets (GNPs) in acqueous suspensions against Staphylococcus aureus to demonstrate the possibility to use graphene based material in environmental applications. In order to study the GNPs toxicity and antimicrobial activity in an in vivo model, we report an antimicrobial study of graphite nanoplatelets on the S. aureus-Caenorhabditis elegans infection model.
Biomolecules, Jan 18, 2020
The manufacturing processes of commercial probiotic strains may be affected in different ways in ... more The manufacturing processes of commercial probiotic strains may be affected in different ways in the attempt to optimize yield, costs, functionality, or stability, influencing gene expression, protein patterns, or metabolic output. Aim of this work is to compare different samples of a high concentration (450 billion bacteria) multispecies (8 strains) formulation produced at two different manufacturing sites, United States of America (US) and Italy (IT), by applying a combination of functional proteomics, metabolomics, and in vivo analyses. Several protein-profile differences were detected between IT- and US-made products, with Lactobacillus paracasei, Streptococcus thermophilus, and Bifidobacteria being the main affected probiotics/microorganisms. Performing proton nuclear magnetic spectroscopy (1H-NMR), some discrepancies in amino acid, lactate, betaine and sucrose concentrations were also reported between the two products. Finally, we investigated the health-promoting and antiaging effects of both products in the model organism Caenorhabditis elegans. The integration of omics platforms with in vivo analysis has emerged as a powerful tool to assess manufacturing procedures.
International Journal of Molecular Sciences
Probiotics exert beneficial effects on host health through different mechanisms of action, such a... more Probiotics exert beneficial effects on host health through different mechanisms of action, such as production of antimicrobial substances, competition with pathogens, enhancement of host mucosal barrier integrity and immunomodulation. In the context of ageing, which is characterized by several physiological alterations leading to a low grade inflammatory status called inflammageing, evidences suggest a potential prolongevity role of probiotics. Unraveling the mechanisms underlying anti-ageing effects requires the use of simple model systems. To this respect, the nematode Caenorhabditis elegans represents a suitable model organism for the study of both host-microbe interactions and for ageing studies, because of conserved signaling pathways and host defense mechanisms involved in the regulation of its lifespan. Therefore, this review analyses the impact of probiotics on C. elegans age-related parameters, with particular emphasis on oxidative stress, immunity, inflammation and protect...
Microorganisms
Lactic acid bacteria (LAB) share several beneficial effects on human organisms, such as bioactive... more Lactic acid bacteria (LAB) share several beneficial effects on human organisms, such as bioactive metabolites’ release, pathogens’ competition and immune stimulation. This study aimed at determining the probiotic potential of autochthonous lactic acid bacteria isolated from carrots. In particular, the work reported the characterization at the species level of four LAB strains deriving from carrots harvested in Fucino highland, Abruzzo (Italy). Ribosomal 16S DNA analysis allowed identification of three strains belonging to Leuconostoc mesenteroides and a Weissella soli strain. In vitro and in vivo assays were performed to investigate the probiotic potential of the different isolates. Among them, L. mesenteroides C2 and L. mesenteroides C7 showed high survival percentages under in vitro simulated gastro-intestinal conditions, antibiotic susceptibly and the ability to inhibit in vitro growth against Salmonella enterica serovar Typhimurium, Listeria monocytogenes, Pseudomonas aeruginosa...
Science of The Total Environment
Environmental Science and Pollution Research
International Journal of Environmental Research and Public Health
Wastewater carries different pathogenic and non-pathogenic microorganisms that can be dispersed i... more Wastewater carries different pathogenic and non-pathogenic microorganisms that can be dispersed in the surrounding environment. Workers who frequent sewage treatment plants can therefore be exposed to aerosols that contain a high concentration of potentially dangerous biological agents, or they can come into direct contact with contaminated material. This can lead to allergies, infections and occupational health-associated diseases. A characterization of biological risk assessment of bioaerosol exposure is necessary. The aim of this study was to evaluate the application of an interdisciplinary method that combines chemical and biological approaches for the analysis of a bioaerosol derived from a wastewater treatment plant (WWTP) situated in Italy. Sampled filters were analyzed by HPLC-MS/MS spectroscopy that searched for different chemical biomarkers of airborne microorganisms. The analytical quantification was compared to the biological cultural method that revealed an underrated m...
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Papers by Daniela Uccelletti