MiR126-targeted-nanoparticles combined with PI3K/AKT inhibitor as a new strategy to overcome melanoma resistance.

Metastatic melanoma poses significant challenges as a highly lethal disease. Despite the success of molecular targeting using BRAFV600E inhibitors (BRAFis) and immunotherapy, the emergence of early recurrence remains an issue and there is the need for novel therapeutic approaches. This study aimed at creating a targeted delivery system for the oncosuppressor microRNA 126 (miR126) and testing its effectiveness in combination with a phosphatidylinositol 3-kinase (PI3K)/ protein kinase B (AKT) inhibitor for treating metastatic melanoma resistant to BRAFis. To achieve this, we synthesized chitosan nanoparticles containing a chemically modified miR126 sequence. These nanoparticles were further functionalized with an antibody specific to the chondroitin sulfate proteoglycan 4 (CSPG4) melanoma marker. After evaluation in vitro, the efficacy of this treatment was evaluated through an in vivo experiment using mice bearing resistant human melanoma. The co-administration of miR126 and the PI3K/AKT inhibitor in these experiments significantly reduced tumor growth and inhibited the formation of liver and lung metastases. These results provide evidence for a strategy to target an oncosuppressive nucleic acid sequence to tumor cells while simultaneously protecting it from plasma degradation. The system described in this study exhibits encouraging potential for the effective treatment of therapy-resistant metastatic melanoma while also presenting a prospective approach for other forms of cancer.

Poly-(lactic-co-glycolic) Acid Nanoparticles Entrapping Pterostilbene for Targeting Aspergillus Section Nigri.

Poly-(lactic-co-glycolic) acid (PLGA) is a biodegradable, biosafe, and biocompatible copolymer. The Aspergillus section Nigri causes otomycosis localized in the external auditory canal. In this research, Aspergillus brasiliensis, a species belonging to the Nigri section, was tested. Coumarin 6 and pterostilbene loaded in poly-(lactic-co-glycolic) acid nanoparticles (PLGA-coumarin6-NPs and PLGA-PTB-NPs) were tested for fungal cell uptake and antifungal ability against A. brasiliensis biofilm, respectively. Moreover, the activity of PLGA-PTB-NPs in inhibiting the A. brasiliensis infection was tested using Galleria mellonella larvae. The results showed a fluorescence signal, after 50 nm PLGA-coumarin6-NPs treatment, inside A. brasiliensis hyphae and along the entire thickness of the biofilm matrix, which was indicative of an efficient NP uptake. Regarding antifungal activity, a reduction in A. brasiliensis biofilm formation and mature biofilm with PLGA-PTB-NPs has been demonstrated. Moreover, in vivo experiments showed a significant reduction in mortality of infected larvae after injection of PLGA-PTB-NPs compared to free PTB at the same concentration. In conclusion, the PLGA-NPs system can increase the bioavailability of PTB in Aspergillus section Nigri biofilm by overcoming the biofilm matrix barrier and delivering PTB to fungal cells.

Biosynthesis of innovative calcium phosphate/hydrogel composites: physicochemical and biological characterisation.

The goal of supporting and directing tissue regeneration requires the design of new, advanced materials, with features like biocompatibility, biodegradability and adequate mechanical properties. Our work was focused on developing a new injectable biomimetic composite material, based on a peptidic hydrogel and calcium phosphates with the aim of mimicking the chemical composition of natural bone tissue. Arg-Gly-Asp-grafted chitosan was used to promote cell adhesion. The obtained composite hydrogel was characterized with differential scanning calorimetry measurements, rheological analysis, field emission scanning electron microscopy, Fourier transform infrared spectroscopy and nuclear magnetic resonance measurements. The biological responsiveness was assessed using the MG-63 human osteoblast cell line.

Extraction of Carotenoids and Fat-Soluble Vitamins from Tetradesmus Obliquus Microalgae: An Optimized Approach by Using Supercritical CO2.

In recent years, great attention has been focused on rapid, selective, and environmentally friendly extraction methods to recover pigments and antioxidants from microalgae. Among these, supercritical fluid extraction (SFE) represents one of the most important alternatives to traditional extraction methods carried out with the use of organic solvents. In this study, the influence of parameters such as pressure, temperature, and the addition of a polar co-solvent in the SFE yields of carotenoids and fat-soluble vitamins from T. obliquus biomass were evaluated. The highest extraction of alpha-tocopherol, gamma-tocopherol, and retinol was achieved at a pressure of 30 MPa and a temperature of 40 °C. It was observed that overall, the extraction yield increased considerably when a preliminary step of sample pre-treatment, based on a matrix solid phase dispersion, was applied using diatomaceous earth as a dispersing agent. The use of ethanol as a co-solvent, under certain conditions of pressure and temperature, resulted in selectively increasing the yields of only some compounds. In particular, a remarkable selectivity was observed if the extraction was carried out in the presence of ethanol at 10 MPa and 40 °C: under these conditions, it was possible to isolate menaquinone-7, a homologous of vitamin K2, which, otherwise, cannot not recovered by using traditional extraction procedures.

Anti-Candida Biofilm Activity of Pterostilbene or Crude Extract from Non-Fermented Grape Pomace Entrapped in Biopolymeric Nanoparticles.

Polymeric nanoparticle-based carriers are promising agents to deliver drugs to cells. Vitis vinifera phenolic compounds are known for their antifungal activity against Candida albicans. The aim of the present study was to investigate the antifungal activity of pterostilbene or crude extracts from non-fermented grape pomace, entrapped in poly(lactic-co-glycolic) acid nanoparticles (NPs), with diameters of 50 and 150 nm, on Candida biofilm. The fluorescent probe coumarin 6 was used to study the uptake of poly(lactic-co-glycolic)acid (PLGA) NPs in planktonic cells and biofilm. The green fluorescent signal of coumarin 6 was observed in Candida biofilm after 24 and 48 hours. Both pterostilbene and crude pomace extract entrapped in NPs exerted a significantly higher anti-biofilm activity compared to their free forms. The entrapment efficiency of both pterostilbene and crude pomace extract in PLGA NPs was ~90%. At 16 µg/mL, pterostilbene loaded in PLGA NPs reduced biofilm formation of 63% and reduced mature biofilm of 50%. Moreover, at 50 µg/mL, the pomace extract loaded in NPs reduced mature biofilm of 37%. These results strongly suggest that PLGA NPs are promising nanodevices for the delivery of antifungal drugs as the crude grape pomace extract, a by-product of white wine making.

Controlled release of 18-ß-glycyrrhetic acid by nanodelivery systems increases cytotoxicity on oral carcinoma cell line.

The topical treatment for oral mucosal diseases is often based on products optimized for dermatologic applications; consequently, a lower therapeutic effect may be present. 18-ß-glycyrrhetic acid (GA) is extracted from Glycirrhiza glabra. The first aim of this study was to test the cytotoxicity of GA on PE/CA-PJ15 cells. The second aim was to propose and test two different delivery systems, i.e. nanoparticles and fibers, to guarantee a controlled release of GA in vitro. We used chitosan and poly(lactic-co-glycolic) acid based nanoparticles and polylactic acid fibers. We tested both delivery systems in vitro on PE/CA-PJ15 cells and on normal human gingival fibroblasts (HGFs). The morphology of GA-loaded nanoparticles (GA-NPs) and fibers (GA-FBs) was investigated by electron microscopy and dynamic light scattering; GA release kinetics was studied spectrophotometrically. MTT test was used to assess GA cytotoxicity on both cancer and normal cells. Cells were exposed to different concentrations of GA (20-500 µmol l-1) administered as free GA (GA-f), and to GA-NPs or GA-FBs. ROS production was evaluated using dichlorodihydrofluorescein as a fluorescent probe. Regarding the cytotoxic effect of GA on PE/CA-PJ15 cells, the lowest TC50 value was 200 µmol l-1 when GA was added as GA-NPs. No cytotoxic effects were observed when GA was administered to HGFs. N-acetyl Cysteine reduced mortality induced by GA-f in PE/CA-PJ15 cells. The specific effect of GA on PE/CA-PJ15 cells is mainly due to the different sensitivity of cancer cells to ROS over-production; GA-NPs and GA-FBs formulations increase, in vitro, this toxic effect on oral cancer cells.

Endocytic pathways involved in PLGA nanoparticle uptake by grapevine cells and role of cell wall and membrane in size selection.

KEY MESSAGE: PLGA NPs' cell uptake involves different endocytic pathways. Clathrin-independent endocytosis is the main internalization route. The cell wall plays a more prominent role than the plasma membrane in NPs' size selection. In the last years, many studies on absorption and cell uptake of nanoparticles by plants have been conducted, but the understanding of the internalization mechanisms is still largely unknown. In this study, polydispersed and monodispersed poly(lactic-co-glycolic) acid nanoparticles (PLGA NPs) were synthesized, and a strategy combining the use of transmission electron microscopy (TEM), confocal analysis, fluorescently labeled PLGA NPs, a probe for endocytic vesicles (FM4-64), and endocytosis inhibitors (i.e., wortmannin, ikarugamycin, and salicylic acid) was employed to shed light on PLGA NP cell uptake in grapevine cultured cells and to assess the role of the cell wall and plasma membrane in size selection of PLGA NPs. The ability of PLGA NPs to cross the cell wall and membrane was confirmed by TEM and fluorescence microscopy. A strong adhesion of PLGA NPs to the outer side of the cell wall was observed, presumably due to electrostatic interactions. Confocal microscopy and treatment with endocytosis inhibitors suggested the involvement of both clathrin-dependent and clathrin-independent endocytosis in cell uptake of PLGA NPs and the latter appeared to be the main internalization pathway. Experiments on grapevine protoplasts revealed that the cell wall plays a more prominent role than the plasma membrane in size selection of PLGA NPs. While the cell wall prevents the uptake of PLGA NPs with diameters over 50 nm, the plasma membrane can be crossed by PLGA NPs with a diameter of 500-600 nm.

Improved stability and efficacy of chitosan/pDNA complexes for gene delivery.

Among polymeric polycations, chitosan has emerged as a powerful carrier for gene delivery. Only a few studies have focused on the stability of the chitosan/DNA complex under storage, although this is imperative for nanomedicinal applications. Here, we synthesized polyelectrolyte complexes at a charge ratio of 10 using 50 kDa chitosan and plasmid (p)DNA that encodes a GFP reporter. These preparations were stable up to 3 months at 4 °C and showed reproducible transfection efficiencies in vitro in HEK293 cells. In addition, we developed a methodology that increases the in vitro transfection efficiency of chitosan/pDNA complexes by 150% with respect to standard procedures. Notably, intracellular pDNA release and transfected cells peaked 5 days following transection of mitotically active cells. These new developments in formulation technology enhance the potential for polymeric nanoparticle-mediated gene therapy.

Designing unconventional Fmoc-peptide-based biomaterials: structure and related properties.

We have recently employed L-amino acids in the lipase-catalyzed biofabrication of a class of self-assembling Fmoc-peptides that form 3-dimensional nanofiber scaffolds. Here we report that using d-amino acids, the homochiral self-assembling peptide Fmoc-D-Phe3 (Fmoc-F*F*F*) also forms a 3-dimensional nanofiber scaffold that is substantially distinguishable from its L-peptide and heterochiral peptide (F*FF and FF*F*) counterparts on the basis of their physico-chemical properties. Such chiral peptides self-assemble into ordered nanofibers with well defined fibrillar motifs. Circular dichroism and atomic force microscopy have been employed to study in depth such fibrillar peptide structures. Dexamethasone release kinetics from PLGA and CS-PLGA nanoparticles entrapped within the peptidic hydrogel matrix encourage its use for applications in drug controlled release.

Liposome-Hydrogel Composites for Controlled Drug Delivery Applications.

Various controlled delivery systems (CDSs) have been developed to overcome the shortcomings of traditional drug formulations (tablets, capsules, syrups, ointments, etc.). Among innovative CDSs, hydrogels and liposomes have shown great promise for clinical applications thanks to their cost-effectiveness, well-known chemistry and synthetic feasibility, biodegradability, biocompatibility and responsiveness to external stimuli. To date, several liposomal- and hydrogel-based products have been approved to treat cancer, as well as fungal and viral infections, hence the integration of liposomes into hydrogels has attracted increasing attention because of the benefit from both of them into a single platform, resulting in a multifunctional drug formulation, which is essential to develop efficient CDSs. This short review aims to present an updated report on the advancements of liposome-hydrogel systems for drug delivery purposes.

Preparation of Peptide-Based Magnetogels for Removing Organic Dyes from Water.

Water pollution by organic dyes represents a major health and environmental issue. Despite the fact that peptide-based hydrogels are considered to be optimal absorbents for removing such contaminants, hydrogel systems often suffer from a lack of mechanical stability and complex recovery. Recently, we developed an enzymatic approach for the preparation of a new peptide-based magnetogel containing polyacrylic acid-modified γ-Fe2O3 nanoparticles (γ-Fe2O3NPs) that showed the promising ability to remove cationic metal ions from aqueous phases. In the present work, we tested the ability of the magnetogel formulation to remove three model organic dyes: methyl orange, methylene blue, and rhodamine 6G. Three different hydrogel-based systems were studied, including: (1) Fmoc-Phe3 hydrogel; (2) γ-Fe2O3NPs dispersed in the peptide-based gel (Fe2O3NPs@gel); and (3) Fe2O3NPs@gel with the application of a magnetic field. The removal efficiencies of such adsorbents were evaluated using two different experimental set-ups, by placing the hydrogel sample inside cuvettes or, alternatively, by placing them inside syringes. The obtained peptide magnetogel formulation could represent a valuable and environmentally friendly alternative to currently employed adsorbents.

Spray Coating of Wood with Nanoparticles from Lignin and Polylactic Glycolic Acid Loaded with Thyme Essential Oils.

Ensuring the longevity of wooden constructions depends heavily on the preservation process. However, several traditional preservation methods involving fossil-based compounds have become outdated because they pose a significant risk to the environment and to human health. Therefore, the use of bio-based and bioactive solutions, such as essential oils, has emerged as a more sustainable alternative in protecting wood from biotic attacks. The entrapment of essential oils in polymeric carrier matrices provides protection against oxidation and subsequent degradation or rapid evaporation, which implies the loss of their biocidal effect. In this work, lignin as well as PLGA nanoparticles containing the essential oils from two different thyme species (Thymus capitatus and T. vulgaris) were applied on beech wood samples using spray coating. The prepared coatings were investigated using FTIR imaging, SEM, as well as LSM analysis. Release experiments were conducted to investigate the release behavior of the essential oils from their respective lignin and PLGA carrier materials. The study found that lignin nanoparticles were more effective at trapping and retaining essential oils than PLGA nanoparticles, despite having larger average particle diameters and a more uneven particle size distribution. An analysis of the lignin coatings showed that they formed a uniform layer that covered most of the surface pores. PLGA nanoparticles formed a film-like layer on the cell walls, and after leaching, larger areas of native wood were evident on the wood samples treated with PLGA NPs compared to the ones coated with lignin NPs. The loading capacity and efficiency varied with the type of essential oil, while the release behaviors were similar between the two essential oil types applied in this study.

Peptide-Based Hydrogels: Template Materials for Tissue Engineering.

Tissue and organ regeneration are challenging issues, yet they represent the frontier of current research in the biomedical field. Currently, a major problem is the lack of ideal scaffold materials' definition. As well known, peptide hydrogels have attracted increasing attention in recent years thanks to significant properties such as biocompatibility, biodegradability, good mechanical stability, and tissue-like elasticity. Such properties make them excellent candidates for 3D scaffold materials. In this review, the first aim is to describe the main features of a peptide hydrogel in order to be considered as a 3D scaffold, focusing in particular on mechanical properties, as well as on biodegradability and bioactivity. Then, some recent applications of peptide hydrogels in tissue engineering, including soft and hard tissues, will be discussed to analyze the most relevant research trends in this field.

Confocal Microscopy Investigations of Biopolymeric PLGA Nanoparticle Uptake in Arabidopsis thaliana L. Cultured Cells and Plantlet Roots.

To date, most endocytosis studies in plant cells have focused on clathrin-dependent endocytosis, while limited evidence is available on clathrin-independent pathways. Since dynamin a is a key protein both in clathrin-mediated endocytosis and in clathrin-independent endocytic processes, this study investigated its role in the uptake of poly-(lactic-co-glycolic) acid (PLGA) nanoparticles (NPs). The experiments were performed on cultured cells and roots of Arabidopsis thaliana. Dynasore was used to inhibit the activity of dynamin-like proteins to investigate whether PLGA NPs enter plant cells through a dynamin-like-dependent or dynamin-like-independent endocytic pathway. Observations were performed by confocal microscopy using a fluorescent probe, coumarin 6, loaded in PLGA NPs. The results showed that both cells and roots of A. thaliana rapidly take up PLGA NPs. Dynasore was administered at different concentrations and exposure times in order to identify the effective ones for inhibitory activity. Treatments with dynasore did not prevent the NPs uptake, as revealed by the presence of fluorescence emission detected in the cytoplasm. At the highest concentration and the longest exposure time to dynasore, the fluorescence of NPs was not visible due to cell death. Thus, the results suggest that, because the NPs' uptake is unaffected by dynasore exposure, NPs can enter cells and roots by following a dynamin-like-independent endocytic pathway.

Peptide-Hydrogel Nanocomposites for Anti-Cancer Drug Delivery.

Cancer is the second leading cause of death globally, but conventional anticancer drugs have side effects, mainly due to their non-specific distribution in the body in both cancerous and healthy cells. To address this relevant issue and improve the efficiency of anticancer drugs, increasing attention is being devoted to hydrogel drug-delivery systems for different kinds of cancer treatment due to their high biocompatibility and stability, low side effects, and ease of modifications. To improve the therapeutic efficiency and provide multi-functionality, different types of nanoparticles (NPs) can be incorporated within the hydrogels to form smart hydrogel nanocomposites, benefiting the advantages of both counterparts and suitable for advanced anticancer applications. Despite many papers on non-peptide hydrogel nanocomposites, there is limited knowledge about peptide-based nanocomposites, specifically in anti-cancer drug delivery. The aim of this short but comprehensive review is, therefore, to focus attention on the synergies resulting from the combination of NPs with peptide-based hydrogels. This review, which includes a survey of recent advances in this kind of material, does not aim to be an exhaustive review of hydrogel technology, but it instead highlights recent noteworthy publications and discusses novel perspectives to provide valuable insights into the promising synergic combination of peptide hydrogels and NPs for the design of novel anticancer drug delivery systems.

Lignin Nanoparticles with Entrapped Thymus spp. Essential Oils for the Control of Wood-Rot Fungi.

After decades of utilization of fossil-based and environmentally hazardous compounds for wood preservation against fungal attack, there is a strong need to substitute those compounds with bio-based bioactive solutions, such as essential oils. In this work, lignin nanoparticles containing four essential oils from thyme species (Thymus capitatus, Coridothymus capitatus, T. vulgaris, and T. vulgaris Demeter) were applied as biocides in in vitro experiments to test their anti-fungal effect against two white-rot fungi (Trametes versicolor and Pleurotus ostreatus) and two brown-rot fungi (Poria monticola and Gloeophyllum trabeum). Entrapment of essential oils provided a delayed release over a time frame of 7 days from the lignin carrier matrix and resulted in lower minimum inhibitory concentrations of the essential oils against the brown-rot fungi (0.30-0.60 mg/mL), while for the white-rot fungi, identical concentrations were determined compared with free essential oils (0.05-0.30 mg/mL). Fourier Transform infrared (FTIR) spectroscopy was used to assess the fungal cell wall changes in the presence of essential oils in the growth medium. The results regarding brown-rot fungi present a promising approach for a more effective and sustainable utilization of essential oils against this class of wood-rot fungi. In the case of white-rot fungi, lignin nanoparticles, as essential oils delivery vehicles, still need optimization in their efficacy.

Surface modification of TiO2 nanoparticles with organic molecules and their biological applications.

In recent years, titanium(IV) dioxide nanoparticles (TiO2NPs) have shown promising potential in various biological applications such as antimicrobials, drug delivery, photodynamic therapy, biosensors, and tissue engineering. For employing TiO2NPs in these fields, their nanosurface must be coated or conjugated with organic and/or inorganic agents. This modification can improve their stability, photochemical properties, biocompatibility, and even surface area for further conjugation with other molecules such as drugs, targeting molecules, polymers, etc. This review describes the organic-based modification of TiO2NPs and their potential applications in the mentioned biological fields. In the first part of this review, around 75 recent publications (2017-2022) are mentioned on the common TiO2NP modifiers including organosilanes, polymers, small molecules, and hydrogels, which improve the photochemical features of TiO2NPs. In the second part of this review, we presented 149 recent papers (2020-2022) about the use of modified TiO2NPs in biological applications, in which specific bioactive modifiers are introduced in this part with their advantages. In this review, the following information is presented: (1) the common organic modifiers for TiO2NPs, (2) biologically important modifiers and their benefits, and (3) recent publications on biological studies on the modified TiO2NPs with their achievements. This review shows the paramount significance of the organic-based modification of TiO2NPs to enhance their biological effectiveness, paving the way toward the development of advanced TiO2-based nanomaterials in nanomedicine.

Self-Assembling Peptide-Based Magnetogels for the Removal of Heavy Metals from Water.

In this study, we present the synthesis of a novel peptide-based magnetogel obtained through the encapsulation of γ-Fe2O3-polyacrylic acid (PAA) nanoparticles (γ-Fe2O3NPs) into a hydrogel matrix, used for enhancing the ability of the hydrogel to remove Cr(III), Co(II), and Ni(II) pollutants from water. Fmoc-Phe (Fluorenylmethoxycarbonyl-Phenylalanine) and diphenylalanine (Phe2) were used as starting reagents for the hydrogelator (Fmoc-Phe3) synthesis via an enzymatic method. The PAA-coated magnetic nanoparticles were synthesized in a separate step, using the co-precipitation method, and encapsulated into the peptide-based hydrogel. The resulting organic/inorganic hybrid system (γ-Fe2O3NPs-peptide) was characterized with different techniques, including FT-IR, Raman, UV-Vis, DLS, ζ-potential, XPS, FESEM-EDS, swelling ability tests, and rheology. Regarding the application in heavy metals removal from aqueous solutions, the behavior of the obtained magnetogel was compared to its precursors and the effect of the magnetic field was assessed. Four different systems were studied for the separation of heavy metal ions from aqueous solutions, including (1) γ-Fe2O3NPs stabilized with PAA, (γ-Fe2O3NPs); (2) Fmoc-Phe3 hydrogel (HG); (3) γ-Fe2O3NPs embedded in peptide magnetogel (γ-Fe2O3NPs@HG); and (4) γ-Fe2O3NPs@HG in the presence of an external magnetic field. To quantify the removal efficiency of these four model systems, the UV-Vis technique was employed as a fast, cheap, and versatile method. The results demonstrate that both Fmoc-Phe3 hydrogel and γ-Fe2O3NPs peptide magnetogel can efficiently remove all the tested pollutants from water. Interestingly, due to the presence of magnetic γ-Fe2O3NPs inside the hydrogel, the removal efficiency can be enhanced by applying an external magnetic field. The proposed magnetogel represents a smart multifunctional nanosystem with improved absorption efficiency and synergic effect upon applying an external magnetic field. These results are promising for potential environmental applications of γ-Fe2O3NPs-peptide magnetogels to the removal of pollutants from aqueous media.

Preparation of Hydrogel Composites Using a Sustainable Approach for In Situ Silver Nanoparticles Formation.

The recognized antibacterial properties of silver nanoparticles (AgNPs) characterize them as attractive nanomaterials for developing new bioactive materials less prone to the development of antibiotic resistance. In this work, we developed new composites based on self-assembling Fmoc-Phe3 peptide hydrogels impregnated with in situ prepared AgNPs. Different methodologies, from traditional to innovative and eco-sustainable, were compared. The obtained composites were characterized from a hydrodynamic, structural, and morphological point of view, using different techniques such as DLS, SEM, and rheological measurements to evaluate how the choice of the reducing agent determines the characteristics of AgNPs and how their presence within the hydrogel affects their structure and properties. Moreover, the antibacterial properties of these composites were tested against S. aureus, a major human pathogen responsible for a wide range of clinical infections. Results demonstrated that the hydrogel composites containing AgNPs (hgel@AgNPs) could represent promising biomaterials for treating S. aureus-related infections.

Biosynthesis of Peptide Hydrogel-Titania Nanoparticle Composites with Antibacterial Properties.

The photoantibacterial properties of titania nanoparticles (TiO2NPs) are attracting much interest, but the separation of their suspension limits their application. In this study, the encapsulation of commercial TiO2NPs within self-assembling tripeptide hydrogels to form hgel-TiO2NP composites with significant photoantibacterial properties is reported. The Fmoc-Phe3 hydrogelator was synthesized via an enzymatic method. The resulting composite was characterized with DLS, ζ-potential, SAXS, FESEM-EDS and rheological measurements. Two different concentrations of TiO2NPs were used. The results showed that, by increasing the TiO2NP quantity from 5 to 10 mg, the value of the elastic modulus doubled, while the swelling ratio decreased from 63.6 to 45.5%. The antimicrobial efficacy of hgel-TiO2NPs was tested against a laboratory Staphylococcus aureus (S. aureus) strain and two methicillin-resistant S. aureus (MRSA) clinical isolates. Results highlighted a concentration-dependent superior antibacterial activity of hgel-TiO2NPs over TiO2NPs in the dark and after UV photoactivation. Notably, UV light exposure substantially increased the biocidal action of hgel-TiO2NPs compared to TiO2NPs. Surprisingly, in the absence of UV light, both composites significantly increased S. aureus growth relative to control groups. These findings support the role of hgel-TiO2NPs as promising biocidal agents in clinical and sanitation contexts. However, they also signal concerns about TiO2NP exposure influencing S. aureus virulence.