Cesar Koppe Grisolia

Background: Ultraviolet (UV) radiation, urban pollution and depletion of cutaneous antioxidant reserves are interconnected drivers of photoaging, matrix metalloproteinase (MMP) activation, mitochondrial dysfunction and dermal extracellular matrix degradation. Plant polyphenols can counteract these processes, but topical efficacy is limited by chemical instability, polarity-driven partitioning and a hostile stratum corneum delivery interface. Here we report a green nanostructured lipid carrier co-loading a phenolic-rich jabuticaba (Plinia cauliflora) peel extract and caffeine (NLC-JC), engineered with Amazonian lipids (Astrocaryum murumuru butter and Orbignya phalerata oil) and stabilized by a Natural Deep Eutectic Solvent (NaDES)-like microenvironment. Methods: UPLC profiling was used to determine the polyphenolic composition of the extract. Physicochemical characterization included hydrodynamic diameter and PDI, ζ-potential, lipid matrix organization (XRD) and morphology (TEM), as well as colloidal stability over 30 days. Cytotoxicity was assessed in SH-4 melanocytes (ISO 10993-5) and in zebrafish (Danio rerio) embryos (96-h IC₅₀, GHS acute hazard threshold). Functional antioxidant and photoprotective activity was evaluated by DPPH radical scavenging, and by attenuation of UVC- and H₂O₂-induced anion superoxide generation, alongside assessment of cellular and nuclear morphology under combined oxidative stress. Results: UPLC profiling revealed a polyphenolic fingerprint dominated by ellagic acid (476.8 ± 19.7 µg g⁻¹), quercetin-3-rhamnoside (90.9 ± 4.6 µg g⁻¹), quercetin (83.7 ± 1.4 µg g⁻¹), cyanidin-3-glucoside (73.0 ± 1.2 µg g⁻¹) and protocatechuic acid (68.2 ± 2.2 µg g⁻¹), bioactives with documented anti-MMP, NF-κB-suppressing and ROS-scavenging activities. Optimized NLC-JC showed a hydrodynamic diameter of 56.1 nm (PDI = 0.131), negative ζ-potential, amorphous lipid matrix (XRD) and spherical morphology (TEM, ≈150–200 nm electron-dense particles), with 30-day colloidal stability. NLC-JC exhibited low cytotoxicity in SH-4 melanocytes (IC₅₀ = 2304 μg mL⁻¹), preserving ≥70% viability across the cosmetically relevant range (ISO 10993-5), and a 96-h LC₅₀ of 2013 μg mL⁻¹ in zebrafish (Danio rerio) embryos, three orders of magnitude above the GHS acute hazard threshold (100 mg L⁻¹). Functionally, NLC-JC produced ≈50% DPPH radical scavenging at concentrations comparable to ascorbic acid, attenuated UVC- and H₂O₂-induced anion superoxide generation and preserved cellular and nuclear morphology under combined oxidative stress. Conclusion: These data position NLC-JC as a safe-by-design, multifunctional dermocosmetic platform combining agro-industrial valorization with mechanism-anchored photoprotection.

With the escalating frequency and intensity of global wildfires driven by climate change, fire retardants (FRs) have become essential tools in wildfire management. Despite their widespread use, the environmental safety of newer FR formulations—particularly in relation to aquatic ecosystems and developmental toxicity—remains insufficiently understood. In particular, their effects on fish embryos, which represent a sensitive and ecologically important life stage, are poorly characterized. This study investigated the acute toxicity of three commercially available FRs—N-Borate, N-Phosphate+, and N-Phosphate-—on early life stages of zebrafish (Danio rerio), based on an OECD 236 Fish Embryo Toxicity (FET) test. Notably, N-Phosphate- FR exhibited significant toxicity with a 96 h LC50 of 60 mg/L (0.0055%), while N-Borate (>432 mg/L, >0.032%) and N-Phosphate+ (>1181 mg/L, >0.08%) showed substantially lower toxicity. Sublethal effects, including reduced yolk sac absorption and yolk sac darkening, were observed across all FRs, highlighting potential developmental disruptions. The elevated toxicity of N-Phosphate- FR likely stems from its surfactant content. These findings reveal variations in the acute toxicity of different FR formulations, emphasizing the need for ecotoxicological assessments to guide the selection of safer FRs for wildfire management and to protect aquatic biodiversity. The results highlight the importance of incorporating developmental endpoints in FR risk assessments and provide foundational data for regulatory decisions regarding FR application near aquatic habitats. Further research is necessary to elucidate the mechanisms underlying observed effects and to evaluate cross-species toxicity at environmentally relevant concentrations.

Non-clinical steps for development, validation and biosafety of new medicines and products comprises studies on cells, proteins, and animals. Herein, we evaluated the toxic activity of antitumoral 2-chloro-N-arylacetamides on eukaryotic dividing cells and animal replacement models. Firstly, the cytotoxicity of chloro (compound 2), bromo (compound 3) and nitro (compound 4) acetamides was analyzed by fluorescent assays in fibroblasts. Next, toxicity was evaluated on Allium cepa meristematic cells and 48h-living Artemia salina larvae. Finally, embryos of Danio rerio (zebrafish) were exposed to the compound 2 (0.14 - 7.2 μg/mL) for 120 h exposure. All arylacetamides were cytotoxic on murine and human fibroblasts, with IC₅₀ values ranging from 1.2 μg/mL (5.6 µM = compound 4 on L-929) to 4.9 μg/mL (24 µM = compound 2 on MRC-5 cells), respectively, and inhibited root growth from 10 to 100 µg/mL, corroborated by mitotic index reduction and cell cycle arrest in interphase (p < 0.05) without clastogenic injuries. Compound 2 showed time- and concentration-dependent killing effects on zebrafish embryos. Its 24 h-acute toxicity at higher concentrations (1.93 and 7.2 μg/mL with 90% and 100% death) corroborated toxicity on aquatic A. salina organisms. After 96 h exposure at 0.52 μg/mL (2.55 µM), almost 100% of the embryos showed more than one lethal/sublethal morphological abnormality (p < 0.05). Then, all arylacetamides showed unspecific toxic effects, mainly the halogenated electrophile chloroacetamide. They present strong antimitotic action on vertebrate and vegetal cells, although such antiproliferative activity does not seem to be directly related to chromosomal damage inductions.

This study explores the development and characterization of lipid nanostructures (NLCs) containing natural deep eutectic solvents (NaDESs) derived from taperebá peel extract (Spondias mombin), a by-product rich in bioactive phenolic compounds, including ellagic acid and quercetin. The taperebá extract exhibited a high polyphenol content (2623 mg GAE/L) and notable antioxidant activity, as demonstrated by DPPH (258 mM TEAC/100 mL) and ABTS (495 mM TEAC/100 mL) assays. NLCs were developed using NaDESs to enhance the stability and bioavailability of the antioxidant compounds. Physicochemical characterization confirmed the formation of stable, nanometric, and monodispersed formulations with efficient encapsulation. Biological evaluation of the NLC-TAP-NaDES formulation demonstrated its remarkable capacity to mitigate oxidative stress in cells subjected to H₂O₂-induced ROS generation. Fluorescence imaging revealed a significant reduction in intracellular ROS levels in treated cells compared to untreated controls, confirming the antioxidant efficacy of the formulation. This outcome underscores the synergy between NaDESs and NLC systems in protecting and delivering phenolic compounds. This study highlights the potential of utilizing underexplored by-products, such as taperebá peels, to develop sustainable and effective antioxidant delivery systems. The NLC-TAP-NaDES platform combines nanotechnology with green chemistry principles, presenting significant implications for the treatment of oxidative stress-related conditions and broader applications in pharmaceutical and nutraceutical sciences. These findings contribute to advancing sustainable innovations in antioxidant therapies, leveraging the dual benefits of bioeconomy and high-performance nanomaterials.

This study explores the development and characterization of lipid nanostructures (NLCs) containing natural deep eutectic solvents (NaDES) derived from taperebá peel extract (Spondias mombin), a by-product rich in bioactive phenolic compounds, including ellagic acid and quercetin. The taperebá extract exhibited a high polyphenol content (2623 mg GAE/L) and notable antioxidant activity, as demonstrated by DPPH (258 mM TEAC/100 mL) and ABTS (495 mM TEAC/100 mL) assays. NLCs were developed using NaDES to enhance the stability and bioavailability of the antioxidant compounds. Physicochemical characterization confirmed the formation of stable, nanometric, and monodispersed formulations with efficient encapsulation. Biological evaluation of the NLC-TAP-NaDES formulation demonstrated its remarkable capacity to mitigate oxidative stress in cells subjected to H₂O₂-induced ROS generation. Fluorescence imaging revealed a significant reduction in intracellular ROS levels in treated cells compared to untreated controls, confirming the antioxidant efficacy of the formulation. This outcome underscores the synergy between NaDES and NLC systems in protecting and delivering phenolic compounds. The study highlights the potential of utilizing underexplored by-products, such as taperebá peels, to develop sustainable and effective antioxidant delivery systems. The NLC-TAP-NaDES platform combines nanotechnology with green chemistry principles, presenting significant implications for the treatment of oxidative stress-related conditions and broader applications in pharmaceutical and nutraceutical sciences. These findings contribute to advancing sustainable innovations in antioxidant therapies, leveraging the dual benefits of bioeconomy and high-performance nanomaterials.

Different phthalate compounds, known as emerging contaminants, such as Di-n-butyl (DBP) and di-iso-pentyl (DiPeP) phthalates, have been found in aquatic environments and are widely used as additives to increase the flexibility of plastics. This study intended to evaluate the toxicity in zebrafish embryos and larvae of DBP and DiPeP, as well as the mixture of the two compounds. Fish embryo toxicity, neurotoxicity, oxidative stress, and genotoxicity studies were carried out in Danio rerio (zebrafish) embryos and larvae. Fish were exposed to DBP and DiPeP in concentrations ranging from 0.001 to 0.125 mg/L and also a combination of compounds DBP + DiPeP (1:1 MIX). DBP caused mortality and embryo malformations from 0.062 mg/L, DiPeP at 0.125 mg/L and the mixture at 0.031 mg/L, indicating the potential toxicity of these phthalates. Isolated phthalates and in mixture induced neurotoxicity and oxidative stress, at low concentrations in zebrafish larvae. No genotoxicity was found through comet assay in larvae. DBP was more toxic than DiPeP, and the mixture was more toxic than both phthalates when tested in isolation, showing some kind of interaction.

The industries are searching for greener alternatives for their productions due to the rising concern about the environment and creation of waste and by-products without industrial utility for that specific line of products. This investigation describes the development of two stable nanostructured lipid carriers (NLCs): one is the formulation of a standard NLC, and the other one is the same NLC formulation associated with a natural deep eutectic solvent (NaDES). The research presents the formulation paths of the NLCs through completeness, which encompass dynamic light scattering (DLS), zeta potential tests, and pH. Transmission electron microscopy (TEM) and confocal microscopy were performed to clarify the morphology. Cytotoxicity tests with zebrafish were realized, and the results are complementary to the in vitro outcomes reached with fibroblast L132 tests by the MTT technique and the zymography test. Infrared spectroscopy and X-ray diffractometry tests elucidated the link between the physicochemical characteristics of the formulation and its behavior and properties. Different cooling techniques were explored to prove the tailorable properties of the NLCs for any industrial applications. In conclusion, the compiled results show the successful formulation of new nanocarriers based on a sustainable, eco-friendly, and highly tailorable technology, which presents low cytotoxic potential.

Iron oxide nanoparticles (FeO-NPs) are widely used in scientific and technological fields. Environmental concerns have been raised about residual FeO-NPs levels as their toxicity and bioaccumulative potential are not well understood. Oreochromis niloticus were exposed to nanoparticles of γ-Fe₂O₃ and Fe₃O_4. Micro-CT 3D image and grayscale graphic assessments revealed the accumulation of radiopaque material in the digestive tract of fish exposed to FeO-NPs. Histological analysis showed the presence of such NPs in the hepatopancreas, gills, kidneys, and muscles. No genotoxicity occurred, through micronucleus test and comet assay in peripheral erythrocytes. Body clearance was confirmed by iron-content reduction in organisms exposed to FeO-NPs after recovery period. No tissue injuries were observed in the exposed animals which may be attributed to the absence or low toxicity of iron oxide nanoparticles under the study conditions. O. niloticus showed tolerance to sublethal exposures to FeO-NPs.