In this study, Cu2+ and Pr3+doped Fluorapatite (FAP) nanoparticles were successfully synthesized via a hydro-thermal method. Their structural, optical, cytotoxic, and antibacterial properties were systematically investigated. X-ray diffraction (XRD) confirmed the preservation of the whole hexagonal fluorapatite structure upon doping and co-doping with Cu2+ and Pr3+ ions, with crystallite sizes estimated using the Scherrer equation as 21.6 nm for pure FAP, 33.7 nm for FAP-Cu, 33.0 nm for FAP-Pr, and 17.9 nm for co-doped FAP-Pr-Cu. The Williamson-Hall (W-H) analysis indicated larger apparent crystallite sizes ranging from 25.2 nm (FAP-Pr-Cu) to 85.5 nm (FAP-Cu), suggesting strain-induced peak broadening in the doped samples. Fourier-transform infrared (FTIR) spectroscopy revealed that doping-induced modifications of vibrational modes of fluorapatite matrix and their corresponding intensities, suggesting small structural alterations. By using X-ray photoelectron spectroscopy (XPS), we have confirmed the incorporation of Cu2+ and Pr3+ into the FAP lattice and verified their respective oxidation states. The elemental composition of the prepared powders was investigated by energy dispersive X-ray spectroscopy (EDX). Scanning electron microscopy (SEM) showed agglomerated particles with a sheet-like morphology, while dynamic light scattering (DLS) measurements indicated an increase in the hydrodynamic size upon doping; the average particle sizes ranged from 171.4 nm for pure FAP to 203.0 nm, confirming the impact of dopant incorporation on particle agglomeration. UV-visible absorption spectroscopy revealed a direct bandgap energy of 4.78 eV for pure FAP, whereas the doped samples exhibited bandgap energies ranging between 4.51 eV and 5.40 eV, depending on the doping element. The antibacterial activity was evaluated against clinical bacterial pathogens implicated in oral cavity disease and vagina infection (S1, S2, S3, S4, E1) using the microdilution method. The results demonstrated that Cu-Pr co-doped FAP exhibited significantly enhanced antibacterial efficacy against staphylococcus aureus (S1) and Enterococcus faecalis (E1), with minimum inhibitory concentration (MIC) of 0.123 and 1.11 mg/mL, respectively. By contrast, pure FAP was found to have a MIC of 1.11 and 3.33 mg/mL for the same bacteria. Additionally, cytotoxicity assessment on the human osteosarcoma cell line (U2OS) was evaluated using the MTT assay, which indicated that the co-doped sample exhibited lower toxicity.

Recurrent brain tumors—including high-grade gliomas, brain metastases, and aggressive meningiomas—continue to carry a poor prognosis, with high mortality despite therapeutic advances. The aim of this narrative review is to summarize and critically discuss the current evidence on the role of intra-arterial radioligand therapy (RLT) in the treatment of recurrent brain tumors. RLT, a targeted form of radionuclide therapy, has gained increasing attention for its potential theranostic applications in neuro-oncology. A literature search was conducted using PubMed and Scopus, including clinical studies evaluating intra-arterial radioligand delivery in central nervous system tumors. Recent research has explored intra-arterial administration of radioligands targeting somatostatin receptors and prostate-specific membrane antigen (PSMA). Somatostatin receptors are overexpressed in meningiomas, while PSMA is highly expressed in the neovasculature of glioblastomas and brain metastases; both targets can be addressed using lutetium-177 (177Lu)- or actinium-225 (225Ac)-labeled radiopharmaceuticals, traditionally delivered intravenously. Available evidence indicates that the intra-arterial route achieves markedly higher radionuclide uptake on 68Ga-PSMA-11 and 68Ga-DOTATOC PET, as well as increased absorbed doses in dosimetric models. Dosimetric analyses consistently show greater tracer accumulation compared with intravenous administration, without evidence of significant peri-procedural toxicity. Uptake in healthy brain tissue is minimal, and no relevant differences have been reported in liver or salivary gland accumulation between intra-arterial and intravenous RLT. Although based on heterogeneous and limited data, intra-arterial RLT appears to be a promising therapeutic strategy for recurrent brain tumors. Future research should focus on improving radioligand delivery beyond the blood–brain barrier and enhancing effective tumor targeting.

Hypothesis. Phase-shift perfluorocarbon microdroplets (MDs) are a promising and flexible method for challenging applications in colloid science, such as customizing chemical reactions or enhancing tumor gene delivery efficiency. To achieve these goals, we focused on stable MDs made of a decafluoropentane core and a dimethyldioctadecylammonium bromide (DDAB) lipid shell that binds DNA through electrostatic interactions to create DNA-MDs assemblies.Experimental. The MDs were obtained via ultrasound homogenization and decorated with complementary DNAs. Fluorescence, confocal microscopy, and dynamic light scattering methodologies were combined to measure the binding features driving DNA-MDs formation and their coalescing interactions. The effect of the DNA-MDs at the cell interface was evaluated by MTT viability assay and DNA transfection in tumor melanoma cells.Findings. We defined the colloidal formulation of DNA-MDs, functional for both chemical and biomedical relevant applications. First, we identified the conditions that enable DNA-MDs to rapidly and selectively coalesce via oligonucleotide hybridization at the MDs interface. Then, we used DNA-MDs to efficiently enter nucleic acids into human melanoma cells, even within a two-step transfection protocol to promote intracellular DNA strand displacement.Our results could shed new light on conducting controlled reactions in small volumes, realizing microreactors, as well as designing consensus gene transfection strategies.

BACKGROUND:Extracellular vesicles (EVs) are stable carriers of molecular signals and can cross the blood-brain barrier, making them promising non-invasive biomarkers for diseases of the central nervous system. While their diagnostic potential is already established and is present in many clinical trials for brain tumors, applications in neurodegenerative disorders such as Parkinson's disease (PD) are still emerging. Although researchers have shown that EVs are involved in the intercellular diffusion of aggregated α-synuclein, a hallmark of PD pathology, the clinical use of EVs as disease biomarkers is still limited by the lack of standardized and selective protocols for EV isolation and analysis. RESULTS:Here, we proposed a dual biochemical and spectroscopic approach to selectively detect PD-associated EVs. As a cellular model of dopaminergic neurons, we employed the human neuroblastoma cell line SH-SY5Y differentiated with retinoic acid and then subjected to 6-hydroxydopamine (6-OHDA) insult to mimic PD-like neurodegeneration. Western blot analysis of SH-SY5Y cells exposed to the neurotoxin revealed high-molecular-weight α-synuclein species consistent with oligomeric forms, which are known to be highly toxic and were also detected in EVs. Moreover, a dot-blot assay with selective sensitivity for aggregated α-synuclein revealed its localization on the membrane surface of fresh, intact PD-like-derived EVs. In addition, Fourier transform infrared spectroscopy identified biochemical signatures that may correlate with pathological states, distinguishing EVs carrying aggregated α-synuclein. SIGNIFICANCE:By combining these biochemical and spectroscopic methods, which required minimal sample volumes, we were able to selectively identify the class of vesicles carrying the aggregated, toxic-form of α-synuclein. As proof of concept, our findings highlight the diagnostic and non-invasive potential of these biomarkers for distinguishing pathology, supporting their possible use in liquid biopsy for neurodegenerative diseases.

Plastics are widely produced due to their stability and ease of manufacturing, but many of them quickly become a waste, breaking down into microplastics and nanoplastics. While methods for the identification and characterization of plastic particles are well consolidated, the small size of nanoplastics presents challenges for their detection and analysis. Furthermore, due to the difficulty of identifying nanoplastics, analytical studies concerning their effect on cells and a comprehensive spectroscopic characterization are still lacking. In this paper, we overcome this obstacle by synthesizing and characterizing, for the first time, PET nanoparticles with specific, stable dimensions through a top-down approach. Using hexafluoroisopropanol-chloroform as a solvent, we prepared PET solutions at various concentrations and analyzed their spectral properties over time. Our results show that PET aggregates into nanoparticles, the quantity of which increases with concentration. These findings provide crucial insights for the detection of nanoplastics in environmental samples through fluorescence measurements and can potentially be used to produce stable PET nanoparticles to evaluate their cytotoxicity.

Herein, we reported a potentiometric pH screen-printed electrode incorporating Carbon Black (CB) nanomaterial and chemically stable pH-sensitive Polyaniline (PANI) for revealing orthopedic infections, as pH is a pivotal biomarker. A dispersion of CB/PANI nanocomposite was easily drop-cast onto the working electrode surface to deliver a straightforward and efficient functionalization of the printed sensor. An in-depth psychochemical and morphological characterization of the nanocomposite was conducted by Dynamic Light Scattering, Electrophoretic Light Scattering, and Transmission Electron Microscopy. 90 % CB - 10 % PANI mixture was selected for electrode functionalization owing to an outstanding dispersibility and an efficient trade-off between linearity in the pH range 3-8 (R2 = 0.994), reliable reproducibility (RSD% = 0.9 %, n = 3), and lack of memory effect, alongside noteworthy sensitivity (-74 ± 3 mV/pH unit). The proposed sensor further showcased a notable selectivity, resulting in negligible pH deviations when exposed to feasible interfering candidates in synovial fluid. Storage stability was also tracked for over 1 month by monitoring the potentiometric response of the designed sensor, kept dry at room temperature. The ANOVA analysis and post-hoc Tukey test demonstrated a stable potential signal, which has been achieved from the second week, with a performance coherence up to one month. Finally, sensor validation was carried out in real samples of healthy and infected synovial fluid by comparing the response of the developed sensor to the pH-meter and the outcome of microbiological analysis, demonstrating the accuracy and the effectiveness of the stable, highly sensitive, and mass-produced printed sensor.

Polymeric microbubbles (MBs) are increasingly being explored as contrast agents for ultrasound (US) imaging and as carriers for US-mediated therapies. Many of these applications benefit from MBs that exhibit strong nonlinear acoustic responses and robust cavitation capabilities. Although structural features, such as size, are known to influence the overall acoustic behavior of MBs, their specific impact on nonlinear responses and cavitation dynamics remains poorly understood and characterized. In this study, we investigated the size-dependent acoustic properties of poly(butyl cyanoacrylate) (PBCA) MBs. Our results show that larger PBCA MBs produce stronger acoustic signals and more pronounced nonlinear responses, including second harmonic generation, whereas smaller PBCA MBs exhibit greater acoustic stability and a higher capacity to sustain stable cavitation. These findings elucidate the role of size in shaping the acoustic behavior of polymeric MBs in a quantitative manner and may inform the design of new agents for enhanced US imaging and US-mediated therapies.

Microbubbles (MB) are widely used as contrast agents for ultrasound (US) imaging and US‐enhanced drug delivery. While the majority of studies utilize commercial MB formulations, increasing experimental evidence indicates that distinct MB features critically determine their diagnostic and therapeutic performance. Here, it is shown that shell stiffness engineering of poly(alkyl cyanoacrylate) (PACA) MB, via introducing monomers with varying alkyl chain lengths and glass transition temperatures, preserves a narrow size distribution ≈2–3 µm, while enhancing MB drug loading, in vitro sonoporation capability, and in vitro and in vivo acoustic responses. All‐atom molecular dynamics simulations and spectroscopic experiments demonstrate that MB shell engineering increases drug diffusion rates in the shell, maximizing the loading capacity of the formulations. Atomic force microscopy demonstrates that the stiffness of the MB shell can be tailored by more than ten‐fold, boosting sonoporation and imaging performance. Altogether, the work provides new insights into the control of polymeric MB structure and performance via dedicated shell engineering, promoting applications in US imaging and therapy.

The establishment of theranostic devices by combining multimodal real-time intraoperative imaging for brain tumor surgery with targeted drug delivery may provide therapeutic advantages for patients with malignant gliomas. Our group has recently developed a new generation of novel microbubbles (MBs), with an air core and a crosslinked poly(vinyl alcohol) shell, called PVA MBs. The PVA MB surface was engineered to support near-infrared (NIR) imaging with a fluorescence probe (C790) for the surgical microscope. The attachment to a cyclic pentapeptide containing the RGD sequence promotes active adhesion and direct targeting of endothelial tumor integrins. The conjugation of temozolomide (TMZ), an alkylating chemotherapy proven to be effective against malignant gliomas, provides a unique therapeutic advantage. The potential toxicity of this novel technology was assessed in rats by intravenous injections of two doses of naked MBs and MBs equipped with RGD for targeting tumor integrins, NIR fluorescence (CF790) for real-time visualization, and TMZ as a cytotoxic component, at two time points, 10 min and 7 days, for potential acute and chronic responses in rats [(1) MB, (2) MB-C790-RGD, and (3) MB-C790-RGD-TMZ]. No mortality occurred during the 7-day study period in any of the dosing groups. Decreased hemoglobin and hematocrit levels and increased triglyceride levels were noticed in the high-dose naked MBs and MBs-CF790-RGD groups. These findings may be associated with their enlarged spleen and liver, observed during necropsy. Histopathology examination in the high-dose animals showed the development of treatment-related changes seen mostly 7 days post dosing, consisting of granulomatous inflammation and foreign body reaction. Accordingly, we concluded that the low-dose tested items appeared to be safe. The results allow us to proceed with planning for an efficacy study before making the first attempt to use this technology in clinical practice.

Background: Intra-arterial cerebral infusion (IACI) of radiotherapeutics is a promising treatment for glioblastoma (GBM) recurrence. We investigated the in silico feasibility and safety of Yttrium-90-Poly(vinyl alcohol)-Microbubble (90Y-PVA-MB) IACI in patients with recurrent GBM and compared the results with those of external beam radiation therapy (EBRT). Methods: Contrast-enhanced T1-weighted magnetic resonance imaging (T1W-MRI) was used to delineate the tumor volumes and CT scans were used to automatically segment the organs at risk in nine patients with recurrent GBM. Volumetric Modulated Arc Therapy (VMAT) treatment plans were generated using a clinical treatment planning system. Assuming the relative intensity of each voxel from the MR-T1W as a valid surrogate for the post-IACI 90Y-PVA-MB distribution, a specific 90Y dose voxel kernel was obtained through Monte Carlo (MC) simulations and convolved with the MRI, resulting in a 90Y-PVA-MB-based dose distribution that was then compared with the VMAT plans. Results: The physical dose distribution obtained from the simulation of 1GBq of 90Y-PVA-MBs was rescaled to ensure that 95% of the prescribed dose was delivered to 95% or 99% of the target (i.e., A95% and A99%, respectively). The calculated activities were A95% = 269.2 [63.6–2334.1] MBq and A99% = 370.6 [93.8–3315.2] MBq, while the mean doses to the target were 58.2 [58.0–60.0] Gy for VMAT, and 123.1 [106.9–153.9] Gy and 170.1 [145.9–223.8] Gy for A95% and A99%, respectively. Additionally, non-target brain tissue was spared in the 90Y-PVA-MB treatment compared to the VMAT approach, with a median [range] of mean doses of 12.5 [12.0–23.0] Gy for VMAT, and 0.6 [0.2–1.0] Gy and 0.9 [0.3–1.5] Gy for the 90Y treatments assuming A95% and A99%, respectively. Conclusions: 90Y-PVA-MB IACI using MR-T1W appears to be feasible and safe, as it enables the delivery of higher doses to tumors and lower doses to non-target volumes compared to the VMAT approach.

Range verification procedures are crucial in carbon-ion radiotherapy (CIRT) to avoid undesired exposure while harnessing the superior features in dose-depth distribution that allow for better biological effectiveness than other adopted radiotherapies. Recently, we have proposed an innovative approach to dosimetry and uncertainties measurement in CIRT by upgrading phase-change ultrasound contrast agents to function as injectable tracers for ionizing particle beams. We developed a stable formulation of superheated nanodroplets (NDs) made of a liquid perfluorobutane core encapsulated within a cross-linked poly-(vinyl alcohol) shell. The core vaporization is triggered by the high-linear energy transfer (LET) carbon ions at the Bragg peak, generating echogenic microbubbles, whereby the resulting echo-contrast relates to beam range with submillimeter precision. So far, in vitro experiments revealed the dependence of the NDs' response to carbon-ion radiation on their size, concentration, as well as particle fluence, which is associated with CIRT dose and beam energy. In this study, we further investigate the biological effect of carbon-ion radiation, before and after the Bragg peak, in 2D cell layer system and correlate this effect with the NDs' radiation response. We evaluated the interaction of nanodroplets of various sizes with the V-79 cell line, a well-established radiobiological model. Tissue-mimicking phantoms containing NDs, along with untreated cells and cells treated with size-sorted NDs, were exposed to C-ion doses of 4 and 8 Gy (130 mm, E = 257.5 MeV/u) at 37 °C. Results indicated that nanodroplets show affinity for the cells and good biocompatibility after incubation. Ultrasound image processing of the vaporization range of nanodroplets, triggered by the carbon-ion radiation, reveals a strong correlation with the cell mortality at the Bragg peak, as assessed by Crystal Violet staining. Our findings highlight the potential of nanodroplets as an injectable and noninvasive dosimeter from the perspective of real-time range verification, offering the ability to optimize treatment plans.

Vascular Endothelial Growth Factor C (VEGFC) is a promising biological drug, with preclinical studies indicating its potential for treating myocardial infarction, neurodegenerative diseases, and lymphedema, a condition that currently lacks curative treatment. While adenoviral VEGFC gene therapy has progressed to phase II studies, its clinical efficacy is limited by rapid immune inactivation. This study explores lignin nanoparticles (LNPs) as an alternative VEGFC delivery system. Lignin was extracted from grape shoot-derived wood biomass via the organosolv method and used to synthesize LNPs. The resulting particles, with a Z-Average of 143.6 nm, a PDI of 0.126 and a zeta potential of -41 ± 8 mV, were thoroughly characterized using dynamic light scattering and microscopic techniques to evaluate their structural and morphological features. VEGFC loading and release profiles were assessed, showing efficient loading and controlled release capabilities. Stability in plasma and cell viability were evaluated using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay to assess biocompatibility and therapeutic potential. Our data indicate that VEGFC is relatively stable on its own, suggesting that the key advantage of LNPs lies in their ability to modulate the release profile rather than protect the protein from degradation/inactivation.

N-Acyl amino acids are biodegradable anionic amphiphilic molecules made up of linear fatty acids as hydrophobic tails and amino acids as polar heads, which are promising for their applicability in different technological fields. In the light of widening their use, a deeper understanding of their interactions with biological membranes is required, especially to further assess their toxicological profile. We investigated the interaction between N-decanoyl amino acid surfactants and phospholipid bilayers as simple in vitro models for biological membranes in comparison to sodium dodecyl sulfate using neutron scattering techniques. The information from small angle neutron scattering (SANS, q range from 0.008 to 0.25 Å-1) focusing on liposome-to-surfactant interactions and neutron reflectivity (NR, Q range measured at three incident angles θ = 0.35, 0.65, and 1.5°) focusing on lipid bilayer-to-surfactant interactions was combined to provide a detailed characterization. All amino acid surfactants (C10-alanine, C10-glycine, C10-leucine, C10-methionine, C10-serine, and C10-proline) exhibited a similar behavior in terms of incorporation in liposomes and lipid removal as well as adsorption profiles in bilayers up to their critical micelle concentration (CMC). Notably, bilayer destabilization occurred for all surfactants (except for C10-serine and C10-alanine) at a concentration between CMC and 2× CMC. Such a result demonstrates the exceptional ability of C10-serine and C10-alanine to integrate into bilayers without disruption up to concentrations as high as ∼3-4× CMC. These findings support the lower cytotoxic effect of C10-serine and C10-alanine surfactants, observed in previous studies, and provide new insights on the mechanism of interaction of N-decanoyl amino acids with lipid membranes.

Low-intensity pulsed ultrasound (LIPUS) is a widely used non-invasive approach with therapeutic purposes since it provides physical stimulation with minimal thermal effects. The skin epithelium is the first barrier of the human body that interfaces with LIPUS and is subjected to the highest intensity. Little is known about the impact of LIPUS on the skin surface. This work investigates the biological effects of one-hour exposure to 1 MHz LIPUS on human keratinocytes HaCaT and tumoral SK-MEL-28 skin cells. Specifically, we evaluated the cellular state immediately after LIPUS treatment by analyzing cytogenetic endpoints and the response of cytoskeleton and cell junction proteins. Herein we demonstrate that LIPUS induces genomic damage as shown by an increase of chromosome malsegregation and a consequent decrease of cellular proliferation. The mechanical stimulus produced by LIPUS is also transmitted to the cytoskeletal compartment, inducing the expression and re-organization of junction proteins (i.e., E-cadherin and Desmosomes) and intermediate filaments (i.e., F-actin and Cytokeratins) with impact on cell morphology and cell adhesion. These in vitro results highlight the different outcomes following the cytogenetic damage and the resilience response exerted by the cytoskeleton upon mechanical stress, laying the foundation for future in vivo investigations.

Common bean (Phaseolus vulgaris L.), one of the most important cultivated legumes, requires a high level of water. It is included among the most sensitive species to climate change; drought and salinity cause a reduction in photosynthesis, metabolic and enzymatic alterations, and oxidative stress. To improve crop tolerance to salt, seed priming and acclimation can be useful tools. To test the salt tolerance of beans, a preliminary screening was undertaken on four cultivars of P. vulgaris (Black Turtle, Cargamanto, Bola Roja, Borlotto) by exposing the seeds to different levels of salinity. The salt-sensitive cultivar Borlotto was chosen for experimental greenhouse trials to study the effects of halopriming and acclimation. Primed and non-primed seeds were sown in non-saline soil and acclimated for 2 weeks; then, the plants were watered with non-saline and saline solutions for 4 weeks. At the end of this growth period, the primed plants showed a marked increase in salt stress tolerance, improving the chlorophyll content, phenolic compounds, and many enzymes’ activities, in turn reducing the effect of salt on growth and fruit production compared to the non-primed controls. In conclusion, halopriming can be considered a useful tool to enhance salinity tolerance in beans and other salt-sensitive crops.

Lippia alba (Mill.) N.E.Br. ex Britton and P. Wilson is used in folk medicine of Central and South America for its biological activities: i.e., antifungal, antibacterial, antiviral, and anti-inflammatory. Based on ethnopharmacological information and the increasing interest in this species, this work aimed to test a possible wide use of its essential oil (EO) in pharmaceutical and horticultural applications. Therefore, we focused the attention on the antioxidant activity of the oil as a possible tool to overcome the oxidative stress in both applications. For this purpose, we have chosen three aggressive breast cancer cell lines and two horticultural species (Solanum lycopersicum L. and Phaseolus acutifolius L.) that are very sensitive to salt stress. We determined the antioxidant activity of L. alba EO through the quantification of phenols and flavonoids. Regarding tomato and bean plants under salt stress, L. alba EO was used for the first time as a seed priming agent to enhance plant salt tolerance. In this case, the seed treatment enhanced the content of phenolic compounds, reduced power and scavenger activity, and decreased membrane lipid peroxidation, thus mitigating the oxidative stress induced by salt. While in breast cancer cells the EO treatment showed different responses according to the cell lines, i.e., in SUM149 and MDA-MB-231 the EO decreased proliferation and increased antioxidant activity and lipid peroxidation, showing high cytotoxic effects associated with the release of lactate dehydrogenase, vice versa no effect was observed in MDA-MB-468. Such antioxidant activity opens a new perspective about this essential oil as a possible tool to counteract proliferation in some cancer cell lines and in horticulture as a seed priming agent to protect from oxidative damage in crops sensitive to salinity.

The Acoustic Droplet Vaporization (ADV) (1), a phase-change of the droplets core from liquid to vapor phase upon ultrasound irradiation, burst renewed interest in droplet emulsions, a traditional topic of colloidal science, opening up innovative applications in biomedicine. Droplets undergoing ADV share similar liquid cores, typically perfluorocarbons (PFCs), however, the nature of the shells can be polymeric (2) or lipidic (3, 4). Such difference imparts to phase-change droplets diverse acoustic and mechanical behaviors (5). In this contribution we present some results concerning a general strategy for the formulation of polymer or lipid shelled submicron droplets. Other key points for the use of ADV-responsive in biomedical applications will be addressed. Recently we have extended the concepts of ADV to radiation responsive droplets for dosimetry in cancer treatment with hadronic radiation. Results on this activity will be also reported (6). References: (1) J. Acoust. Soc. Am. (2004) 116 (1): 272–281; (2) Chem. Commun., 2013, 49, 5763; (3) JoVE, 169 (2021); (4) Langmuir (2019), 35, 10116–10127; (5) Phys. Chem. Chem. Phys., 2016, 18, 8378; (6) COCIS, 49, 118–132 (2020).