Toxicity Studies Research Articles

Low-melting mixture solvents as novel solubilizing agents: Improving tolvaptan formulation and therapeutic efficacy

Presently, a significant number of pharmaceutical drugs exhibit poor solubility in water and fall within categories II and IV of the biopharmaceutical classification system. This predicament has prompted the exploration for novel solvents or excipients to enhance the solubility and bioavailability of the active compounds in aqueous environments. Low-melting mixture solvents (LoMMSs) have gained recognition as potential solvents for active pharmaceutical ingredients to improve therapeutic efficacy and delivery. This study focuses on the preparation of a eutectic mixture composed of betaine, malic acid, proline, and water (BE-MA-PR-WA) to enhance the formulation options for tolvaptan, an FDA-approved medication used to treat acute congestive heart failure, which has limited solubility in aqueous solutions. The findings presented in this article represent the first reported results on the administration of LoMMS-solubilized tolvaptan. Furthermore, an examination of the solubility and bioavailability of tolvaptan utilizing this mixture was conducted, alongside an assessment of its toxicity. The solubility of tolvaptan in BE-MA-PR-WA (1:1:1:7) increased 483 times compared to solubility in water. Toxicity studies indicated that the LoMMS formulation BE-MA-PR-WA was safe for oral administration. The oral bioavailability of tolvaptan was significantly increased by 1.9-fold in the BE-MA-PR-WA than that in water. In addition, the 5-hour urine volume of rats indicated that BE-MA-PR-WA also enhanced the diuretic effects of tolvaptan in rats. The findings suggest that BE-MA-PR-WA enhance the solubility and bioavailability of tolvaptan, and are deemed non-toxic. Consequently, LoMMS may be deemed promising solubilizing agents, potentially facilitating the absorption of poorly bioavailable medications.

Synthesis, molecular docking, drug likeness, in silico toxicity and DFT studies of small molecules as p53-MDM2 interaction and COX-2 dual inhibitors

Synthesis, molecular docking, drug likeness, in silico toxicity and DFT studies of small molecules as p53-MDM2 interaction and COX-2 dual inhibitors

Antidiarheal Activity of Methanol Seed Extract of Cola Acuminata Schott & Endl (Malvaceae) In Mice

Cola acuminata nut extract has been shown to relax smooth muscles of the intestine in vitro, and this has made the plant popular as a potential antidiarrheal agent. This study aimed at evaluating the antidiarrheal activity of methanolic seed extract of Cola acuminata in mice. The extract was subjected to phytochemical analysis. The acute toxicity (LD50) study was conducted according to Lorke's method. The antidiarrheal activity was evaluated using castor oil-induced diarrhea and charcoal meal method in mice. Groups of mice were orally administered water (10 ml/kg), charcoal, Cola acuminata nut extract (150, 300 and 600 mg/kg), loperamide 2mg/kg, and atropine 1 mg/kg before castor oil. Phytochemical screening revealed the presence of alkaloids, cardiac glycosides, tannins, triterpenes, and saponins. The oral LD50 of the extract was estimated to be 2154 mg/kg. The extract produced a significant p < 0.05 increase in time of onset of diarrhea compared to the distilled water group. The extract produced dose dependent inhibition of the number of wet faeces. Extract at 600 mg/kg had more protection against diarrhea than the Loperamide. The extract did not significantly (p > 0.05) affect the intestinal movement of charcoal meal.

Molecular Docking, Molecular Dynamics, pkCSM Drug‐Likeness Profiles, Toxicity, and DFT Study of the Antioxidant and Anticancer Activities of Three Flavonoid Derivatives

AbstractThis work presents a theoretical study of the antioxidant and anticancer properties of three flavonoid derivatives Isoquercitin, Ophioglonol, and Quercetin. DFT calculations were performed to analyze their electronic properties, providing insights into their reactivity and mechanism of action. Molecular docking and molecular dynamics simulations were conducted to examine the stability behavior of the protein‐ligand complexes, providing a detailed mechanism of the interactions at the atomic level. Additionally, some physicochemical and pharmacokinetic predictions were examined by using several web servers. Quercetin and isoquercetin show therapeutic properties, with MESP plots revealing interaction regions with SOD and hAPN targets. However, isoquercetin raises concerns related to AMES toxicity and hERG inhibition. The results of Molecular docking and MD simulations provide theoretical guidance for developing effective inhibitors to treat cancer, but further experimental validation is needed to optimize therapeutic efficacy. The MM/GBSA show that Ophioglonol has the strongest binding affinity with the protein, driven by many interactions. The Actionon also demonstrates strong binding, while Isoquercitin and Quercetin have moderate to weaker affinities. The PCA revealed distinct conformational changes in the protein upon binding with the ligand, highlighting specific regions of flexibility, which could be crucial for understanding their binding mechanisms and potential therapeutic effects.

Investigating business toxic leadership

This manuscript investigates toxic leadership in modern business. It focuses on the key role that poor ethics, and Machiavellianism, Narcissism, and Psychopathy (the three elements of toxic leadership (Nahavandi, 2016)) have played in the failures of a variety of modern major business enterprises. Toxic leadership is management that goes beyond corruption and results in irreparable damage to the organization and its stakeholders (e.g., Adolf Hitler). The emphasis is on toxic leadership that has contributed to the catastrophic failures of several major firms over the past quarter century. It uses a book by Stephen Arbogast entitled "Resisting Corporate Corruption (RCC): Practical Cases in Business Ethics from Enron. The manuscript complements this book with a New Leadership Model that enhances the analysis of a variety of recent toxic leadership case studies. The model is illustrated in failures such as the Madoff Ponzi Scheme and in major firms such as Theranos, Nikola, We Work and Volkswagen. It provides a useful tool and shows that the dark side of leadership can be examined in real-world cases. The model is recommended for firms to employ in finding toxic leaders and in evaluating the consequences of their nefarious actions.

Toxicity study of Stachybotrys chartarum (CASRN 67892-26-6) administered by inhalation to B6C3F1/N mice.

Stachybotrys chartarum, also known as "black mold," is a cellulolytic saprophyte with a worldwide distribution. Public concern for potential illnesses associated with water-damaged indoor environments has been heightened since the report of pediatric acute idiopathic pulmonary hemorrhage/hemosiderosis cases in the United States and following recent natural disasters. Although mycotoxicosis and pulmonary immunological endpoints have been previously examined, the systemic toxicity following subchronic inhalation of viable S. chartarum remains undercharacterized. To evaluate the toxicological responses to S. chartarum, B6C3F1/N mice were exposed to viable S. chartarum conidia (spores) twice a week for 3 months. All in-life procedures, including inhalation exposure, test article preparation, and hematology analysis, were completed by the National Institute for Occupational Safety and Health (NIOSH, Morgantown, WV). Battelle (Columbus, OH) conducted terminal necropsies, measured terminal body and organ weights, and evaluated gross lesions onsite at the NIOSH facility. Tissue processing and histopathology were completed at Battelle. Genetic toxicology studies on mouse peripheral blood erythrocytes were conducted by Integrated Laboratory Systems, LLC (Research Triangle Park, NC). (Abstract Abridged).

Acute oral toxicity and genotoxicity assessment of the essential oil from Croton pulegiodorus Baill (Euphorbiaceae) leaves in mice

Essential oils obtained from Croton pulegiodorus leaf are renowned for their biological activities; however, data on their toxicity are limited. Therefore, this study aimed to evaluate the acute oral toxicity and genotoxicity of a C. pulegiodorus leaf essential oil (CPLEO). Chemical characterization of CPLEO was conducted by gas chromatography coupled to mass spectrometry (GC-MS). In vitro assay was performed to verify the hemolytic capacity of the oil in mice erythrocytes. Next, an acute oral toxicity study was conducted on female mice at CPLEO doses of 2000, 1000, 500, 250, 100, and 50 mg/kg. Hematological, biochemical, and histopathological markers were assessed in mice from groups were no death occurred. Relative consumption of water and food and the weight of animals and their organs were also recorded. Finally, a genotoxicity analysis was performed using the micronucleus and comet assays. The extraction yield of CPLEO was 1.149% and its major compounds were ascaridole (23.18%), eucalyptol (17.20%), camphor (14.20%), p-cymene (7.91%), α-terpineol (4.69%), and isobornyl acetate (4.57%). CPLEO showed a hemolytic effect only at high concentrations (185.5–1000 mg/mL). It showed acute oral toxicity in mice with a LD50 of 460.42 mg/kg. CPLEO (50–250 mg/kg) caused some significant changes in hematological and biochemical parameters. Histopathological evaluation indicated alterations in liver and kidneys but transaminases, urea and creatinine levels remained like the negative control. CPLEO administration impaired weight gain and reduced water and food consumption. Finally, it was not genotoxic by both comet and micronucleus tests. The results highlight the need for attention when choosing doses to evaluate the bioactivities of CPLEO.

Safety evaluation of the food enzyme endo-1,3(4)-β-glucanase from the non-genetically modified Talaromyces versatilis strain PF8.

The food enzyme endo-1,3(4)-β-glucanase (3-(1-3;1-4)-β-d-glucan 3(4)-glucanohydrolase; EC 3.2.1.6) is produced with the non-genetically modified Talaromyces versatilis strain PF8 by Erbslöh Geisenheim AG. The food enzyme was free from viable cells of the production organism. It is intended to be used in four food manufacturing processes. Dietary exposure to the food enzyme-total organic solids (TOS) was calculated to be up to 0.110 mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests did not indicate a safety concern. The systemic toxicity was assessed by means of a repeated dose 90-day oral toxicity study in rats. The Panel identified a no observed adverse effect level of 2229 mg TOS/kg bw per day, the highest dose tested, which when compared with the estimated dietary exposure resulted in a margin of exposure of at least 20,264. A search for homology of the amino acid sequence of the food enzyme to known allergens was made and four matches with respiratory or contact allergens were found. The Panel considered that the risk of allergic reactions upon dietary exposure cannot be excluded, but the likelihood is low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns, under the intended conditions of use.

Dermal toxicity studies of 1,2-benzisothiazolin-3-one (CAS number: 2634-33-5) in Sprague-Dawley rats

Biocides are used as preservatives in various household products, and 1,2-benzisothiazolin-3-one (BIT) is one of the popular chemicals. Therefore, BIT is highly likely to be exposed to human skin, necessitating dermal toxicity evaluation. In this study, we aimed to investigate dermal toxicity, eyes and skin irritation, and skin sensitization of BIT. All studies were conducted according to the Organisation for Economic Co-operation and Development testing guidelines. In acute dermal toxicity using rats, no treatment-related responses were observed at the highest dose (2000 mg/kg). A 28-day repeated dermal toxicity study at 1, 4, and 12 mg/kg/day showed transient local skin irritation lesions, including erythema, exfoliation, and crust formation. Based on no systemic effects, the no observed adverse effect level (NOAEL) of BIT of the 28-day repeated dermal toxicity study was determined to be 12 mg/kg/day. Eye and skin irritation tests showed that BIT is a strong irritant and corrosive to the eyes and a mild irritant to the skin. However, BIT showed no skin sensitization reactions in a local lymph node assay. These dermal toxicity studies can provide valuable information for the risk assessment of BIT.

Development of a B‐risk

Abstract In specific contract No 12 issued under the framework agreement OC/EFSA/AMU/2019/02, EFSA requested Open Analytics to implement a web application to do a risk assessment for honey bees, solitary bees and bumble bees. The software is developed in R and consists of a WEB‐based tool composed by several modules providing data entry for active substances, uses, metabolites and the modelling of toxicity studies. The application is developed in a modular form such that new modules can be added when available, either by the developers of the application or by EFSA.

Comprehensive evaluation of the nephrotoxicity of carbon quantum dots: Effects of the surface charge

Carbon quantum dots (CQDs) are garnering attention for their broad applications. This study offers a detailed evaluation of the biomedical safety and health risks of carbon quantum dots (CQDs) with different surface modifications, addressing a key gap in their safe application. It focuses on three CQD types: diammonium citrate-based (CQDs-A), spermidine trihydrochloride-based (CQDs-S), and a combination (CQDs-A/S), analyzing their physicochemical properties, cytotoxicity, oxidative stress, inflammatory responses, and nephrotoxicity. While all CQDs were under 10 nm, their biological impacts varied. Positively charged CQDs-S and CQDs-A/S showed significant cytotoxicity in HEK293 cells, inducing oxidative stress but not activating NLRP3 inflammasome, indicating a limited inflammatory response. Renal integrity remained unaffected, with stable zonula occludens 2 expression and unaltered renal markers. In vivo studies in BALB/c mice further supported the safety of CQDs, showing no organ damage or kidney pathology at high doses. The findings underscore the potential for safe biomedical use of CQDs, particularly when their retention time is minimized. This research makes a novel contribution by linking CQDs' surface charge to cytotoxic effects and oxidative stress, providing key insights into their safe use in biomedicine and filling a critical gap in nanoparticle toxicity studies.

Lack of genotoxicity and subchronic toxicity in safety assessment studies of Akkermansia muciniphila formulation

A powder formulation of viable Akkermansia muciniphila bacteria (AMUC) was evaluated in a 90-day repeated-dose toxicity study in rats and a battery of genotoxicity studies to evaluate AMUC as a food ingredient. All studies followed Organisation for Economic Co-operation and Development protocols (OECD TG 408, 471 473, 474). AMUC was administered to rats via gavage at 0, 500, 1000, and 2000 mg/kg body weight/day (equivalent to 0, 4.1 × 1010, 9.2 × 1010, and 1.64 × 1011 CFU/kg body weight/day). No mortality or treatment-related adverse effects were reported in any endpoints that were attributed to AMUC consumption. No bacterial translocation of viable A. muciniphila from the intestinal tract was found to the liver, mesenteric lymph nodes, or blood. The no-observed-adverse-effect level was concluded to be the highest dose tested (2000 mg/kg body weight/day), approximately 1.64 × 1011 CFU/kg body weight/day. AMUC (nonviable) was not mutagenic when examined in an in vitro bacterial reverse mutation assay and not clastogenic in an in vitro mammalian chromosomal aberration test. Viable AMUC was not genotoxic when evaluated in an in vivo mammalian cell micronucleus assay when administered at up to 1.64 ×1011 CFU/kg body weight/day. These results confirm that AMUC is not toxic under the conditions of these studies.

Non-targeted Screening and Toxicity Study of Safety Risk Substances in Facial Skincare Products: Molecular Networking and Computational Toxicology Strategy

Non-targeted Screening and Toxicity Study of Safety Risk Substances in Facial Skincare Products: Molecular Networking and Computational Toxicology Strategy

Safety evaluation of the food enzyme endonuclease from the non-genetically modified Penicillium citrinum strain NP 11-15.

The food enzyme endonuclease (Aspergillus nuclease S1; EC 3.1.30.1) is produced with the non-genetically modified Penicillium citrinum strain NP 11-15 by Shin Nihon Chemical Co., Ltd. The food enzyme is free from viable cells of the production organism. It is intended to be used in the processing of yeast and yeast products. Dietary exposure to the food enzyme-total organic solids (TOS) was estimated to be up to 0.006 mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests did not indicate a safety concern. The systemic toxicity was assessed by means of a repeated dose 90-day oral toxicity study in rats. The Panel identified a no observed adverse effect level of 1010 mg TOS/kg bw per day, the highest dose tested, which when compared with the estimated dietary exposure, resulted in a margin of exposure of at least 168,333. A search for homology of the amino acid sequence of the food enzyme to known allergens was made and no match was found. The Panel considered that the risk of allergic reactions by dietary exposure cannot be excluded, especially for individuals allergic to Penicillium. However, the likelihood of such reactions will not exceed the likelihood of allergic reactions to Penicillium. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use.

Unveiling the 4-aminoquinoline derivatives as potent agents against pancreatic ductal adenocarcinoma (PDAC) cell lines

Common antimalarials such as artemisinins, chloroquine and their derivatives also possess potent anti-inflamantory, antiviral and anticancer properties. In the search for new therapeutics to combat difficult-to-treat pancreatic carcinomas, we unveiled that 4-aminoquinoline derivatives, with significant antiplasmodial properties and a great safety profile in vivo, have remarkable anticancer activity against pancreatic ductal adenocarcinoma (PDAC) and considerable efficacy in the xenograft model in vivo. The aim of the present study was to further investigate anticancer properties of these compounds in a drug-repurposing manner. The compounds showed profound cytotoxic effects at nanomolar to low micromolar concentration in 2D cultured cells (in vitro) and in the zebrafish PDAC xenograft model (in vivo). A deeper insight into their mechanisms of cytotoxic action showed these compounds induce apoptosis while increasing reactive oxygen species levels along with autophagy inhibition. Additional investigation of the autophagy modulation proved that tested quinoline derivatives cause P62 and LC3-II accumulation in PDAC cells alongside lysosomal alkalinization. Further, in vivo toxicity studies in the zebrafish model showed low toxicity without developmental side effects of the investigated 4-aminoquinolines, while the applied compounds effectively inhibited tumor growth and prevented the metastasis of xenografted pancreatic cells. Taken together, these results highlight the 4-aminoquinolines as privileged structures that ought to be investigated further for potential application in pancreatic carcinoma treatment.

Statement complementing the EFSA Scientific Opinion on application (EFSA-GMO-NL-2015-126) for authorisation of food and feed containing, consisting of and produced from genetically modified soybean MON 87705 × MON 87708 × MON 89788.

Following a request from the European Commission, the GMO Panel assessed additional information related to the application for authorisation of food and feed containing, consisting of and produced from genetically modified soybean MON × MON 87708 × MON 89788 (EFSA-GMO-NL-2015-126). The applicant conducted a 90-day feeding study on GM soybean MON 87705 and provided a proposal for post-market monitoring considering the altered fatty acid profile of GM soybean MON 87705 × MON 87708 × MON 89788, to fulfil the deficiencies identified by EFSA GMO Panel, addressing elements that remained inconclusive from a previous EFSA scientific opinion issued in 2020. The GMO Panel concludes that the 90-day feeding study on GM soybean MON 87705 is in line with the requirements of Regulation (EU) No 503/2013 and that no treatment-related adverse effects were observed in rats after feeding diets containing soybean MON 87705 meals at 30% or 15% for 90 days. The GMO Panel reiterates the recommendation for a PMM for food in accordance with Regulation (EC) No 1829/2003 and Regulation (EU) No 503/2013 and concludes that the proposal provided by the applicant is in line with the recommendations described for the PMM plan of soybean MON 87705 × MON 87708 × MON 89788 in the adopted scientific opinion. Taking into account the previous assessment and the new information, the GMO Panel concludes that soybean MON 87705 × MON 87708 × MON 89788, as assessed in the scientific opinion on application EFSA-GMO-NL-2015-126 and in the supplementary toxicity study, is as safe as its non-GM comparator and the non-GM reference varieties tested and does not represent a nutritional concern in humans and animals, within the scope of this application.

Fumaric acid per se and in combination with methotrexate arrests inflammation via moderating inflammatory and oxidative stress biomarkers in arthritic rats

Objective: Fumaric acid is a dicarboxylic acid that belongs to the phenolic class enriched in fruits and vegetables that are traditionally used for the treatment of various ailments. The research was planned to find out the anti-inflammatory and anti-arthritic activities of fumaric acid using in-vitro and in-vivo assays. Moreover, safety study was also done. Materials and methods: The 0.1 ml complete Freund’s adjuvant was injected in left hind paw in all Wistar rats except normal rats at day 1 to induced arthritis. The treatment with fumaric acid at 10, 20, 40, and fumaric acid 40 mg/kg together with methotrexate (MTX) was administered to immunized rats at 8th day via oral gavage and continued till 28th day though, MTX was administered as standard control. Results: The fumaric acid notably (p < 0.0001) lessened the paw edema and arthritic scoring, reinstated body and immune organ weight, and oxidation status in treated rats. Fumaric acid notably restored altered C-reactive protein, rheumatoid factor, liver function tests, ESR, WBCs, RBCs and Hb levels in treated rats. The fumaric acid in combination noticeably (p < 0.01–0.0001) suppressed the expression of TNF- α, IL-6, IL-1β, NF-kβ, and COX-2, and over expressed IL-4, and IL-10 in contrast to other treated groups. Fumaric acid had presented a dose-dependent antioxidant, anti-inflammatory and anti-arthritic activities while notable activity exhibited by fumaric acid in combination with MTX. The fumaric acid exhibited non-significant clinical signs of toxicity and mortality in acute toxicity study. The LD50 was more than 2000 mg/kg. Conclusion: Fumaric acid in combination can be used as disease-modifying anti-rheumatic drug but it will need extensive pre-clinical and clinical studies.

Evaluation of Albumin Denaturation, and Phospholipase A2 (Pla2) Inhibition, and Phytochemical Composition of Methanol Leaf Extract of Psychotria Vogeliana, Benth (Rubiaceae)

Inflammation is a protective response elicited by physical, chemical, biochemical, and/or microbial assault on the body, which serves to prevent progression/escalation of injury. If the response is sufficient the body continues to operate normally, if the reaction is excessive a disease condition (inflammation) occurs which predisposes to various health disorders. Current treatment of inflammation includes immunosuppressant, steroidal, and nonsteroidal agents. Most of these are known to have sometimes serious side/adverse effects. Natural compounds from plants known to have pharmacological activities, including anti-inflammatory and are being used as alternatives. This study aimed to investigate the phytochemical composition and anti-inflammatory properties of the methanol leaf extract of Psychotria vogeliana. Acute toxicity study was conducted following the OECD guidelines 423 Annex 2C. Anti-inflammatory activity was evaluated using albumin denaturation, and phospholipase A2 (PLA2) inhibition assay. Phytochemical analysis was conducted using spectrophotometric methods, and GC-FID. The LD50 of the extract is more than 5000 mg/kg. The extract contained bioactive compounds such as catechin, epicateechin, resvesterol, rutin, narigenin, kaempferol, flavonones, tannins, steroids and anthocynins. Anti-inflammatory evaluation showed activity comparable to that of diclofenac. Possible mechanism of action was identified as Phospholipase A2 (PLA2) inhibition. The use of Psychotria vogeliana in traditional medicine for the management and treatment of inflammatory disorders is therefore justified.

Green synthesis of neuroprotective spirocyclic chalcone derivatives and their role in protecting against traumatic optic nerve injury

For clinically prevalent traumatic optic neuropathy (TON) and other retinal and optic nerve injuries lacking effective therapeutic agents, there is an urgent clinical demand for developing highly efficient and safe neuroprotective agents. Here, we have integrated naturally sourced chalcone with isatin through a catalyst-free green synthesis method, reporting a series of spirocyclic chalcone derivatives with significantly lower cytotoxicity than chalcone itself. Following in vitro cell protection assays in models of hydrogen peroxide and glutamic acid-induced damage, multiple active compounds capable of combating both forms of damage were identified. Among these, candidate compound X38 demonstrated promising neuroprotective prospects: in vitro, it attenuated glutamate-induced cell apoptosis, while in vivo, it effectively ameliorated retinal thinning and loss of optic nerve electrophysiological function induced by optic nerve injury. Preliminary mechanistic studies suggest that X38 exerts its neuroprotective effects by mitigating intracellular ROS accumulation, inhibiting JNK phosphorylation, and alleviating oxidative stress. Additionally, acute toxicity studies (intraperitoneal injection, 500 mg/kg) underscored the favorable in vivo safety profile of X38. Taken together, this study has designed a class of safe, neuroprotective spirocyclic chalcone derivatives that can be synthesized using green methods, offering an attractive candidate for treating retinal and optic nerve injuries.

Exploring 99mTc-Labeled Iron-Binding Glycoprotein Nanoparticles as a Potential Nanoplatform for Sentinel Lymph Node Imaging: Development, Characterization, and Radiolabeling Studies.

Lactoferrin, an iron binding glycoprotein-based nanoparticle, has emerged as a promising platform for drug delivery and imaging. This study presents the potential use of the protein nanocarrier in tracking sentinel lymph nodes for cancer staging. Lactoferrin nanoparticles (LF-NPs) were synthesized using a thermal treatment process and optimized to obtain 60-70 nm particle size with PDI less than 0.2. The NPs were characterized microscopically and spectroscopically, ensuring a comprehensive understanding of their physicochemical properties. The LF-NPs were found to be stable in different pH conditions. Their biocompatibility was confirmed through cytotoxicity assessments on RAW 264.7 cells, and hemolysis assay and in vivo toxicity study reveal their safe profile. Additionally, LF-NPs were successfully radiolabeled with technetium-99m (>90% labeling yield). Cell uptake studies with RAW 264.7 exhibited an uptake of ∼6%. Biodistribution studies in Wistar rats shed light on their in vivo behavior and suitability for targeted drug delivery systems. These findings collectively emphasize the multifaceted utility of LF-NPs, positioning them as a promising platform for diverse biomedical innovations.