Uncovering the Fate and Risks of Intravenously Injected Prussian Blue Nanoparticles in mice by an Integrated Methodology of Toxicology, Pharmacokinetics, Proteomics, and Metabolomics.

BACKGROUND: Prussian blue (PB) nanoparticles (NPs) have been intensively investigated for medical applications, but an in-depth toxicological investigation of PB NPs has not been implemented. In the present study, a comprehensive investigation of the fate and risks of PB NPs after intravenous administration was carried out by using a mouse model and an integrated methodology of pharmacokinetics, toxicology, proteomics, and metabolomics. RESULTS: General toxicological studies demonstrated that intravenous administration of PB NPs at 5 or 10 mg/kg could not induce obvious toxicity in mice, while mice treated with a relatively high dose of PB NPs at 20 mg/kg exhibited loss of appetite and weight decrease in the first two days postinjection. Pharmacokinetic studies revealed that intravenously administered PB NPs (20 mg/kg) underwent fast clearance from blood, highly accumulated in the liver and lungs of mice, and finally cleared from tissues. By further integrated proteomics and metabolomics analysis, we found that protein expression and metabolite levels changed significantly in the liver and lungs of mice due to the high accumulation of PB NPs, leading to slight inflammatory responses and intracellular oxidative stress. CONCLUSIONS: Collectively, our integrated experimental data imply that the high accumulation of PB NPs may cause potential risks to the liver and lungs of mice, which will provide detailed references and guidance for further clinical application of PB NPs in the future.

Nanoprotection Against Retinal Pigment Epithelium Degeneration via Ferroptosis Inhibition.

Lethal oxidative stress and ferrous ion accumulation-mediated degeneration/death in retinal pigment epithelium (RPE) exert an indispensable impact on retinal degenerative diseases with irreversible visual impairment, especially in age-related macular degeneration (AMD), but corresponding pathogenesis-oriented medical intervention remains controversial. In this study, the potent iron-binding nanoscale Prussian blue analogue KCa[FeIII (CN)6 ] (CaPB) with high biocompatibility is designed to inhibit RPE death and subsequently photoreceptor cell degeneration. In mice, CaPB effectively prevents RPE degeneration and ultimately fulfills superior therapeutic outcomes upon a single intravitreal injection: significant rescue of retinal structures and visual function. Through high-throughput RNA sequencing and sophisticated biochemistry evaluations, the findings initially unveil that CaPB nanoparticles protect against RPE degradation by inhibiting ferroptotic cell fate. Together with the facile, large-scale preparations and in vivo biosafety, it is believed that the synthesized CaPB therapeutic nanoparticles are promising for future clinical treatment of diverse retinal diseases involving pathological iron-dependent ferroptosis, including AMD.

Complexions therapy and severe intoxication by Thallium salts.

The aim of this paper is to study the clinical features of severe intoxications with thallium salts and developing effective care schemes for the application of potassium hexacyanoferrate (II) and deferasirox for correction of detected disorders. A total of 39 patients diagnosed with severe thallium salt poisoning were examined in two groups. Group I comprised 20 patients with severe thallium salt poisoning, who were prescribed with potassium-iron hexacyanoferrate in a dose of 250 mg/kg/day per os, intravenous potassium infusions, furosemide intravenously in amount of 40 mg three times per, and hemodialysis until the thallium level in the blood dropped below 10 mg/L, lactulose 30 mL two times per day per os. Group II consisted of 19 people with severe thallium salt poisoning, which in addition to the above treatment, received Deferasirox in a dosage of 500 mg two times per day per os. The clinical picture of severe poisoning with thallium salts is characterized by lesions of the gastrointestinal tract, nervous system (central and peripheral), alopecia, heart rhythm disorders, and myocardial ischemia zones. Extension of standard therapy with potassium-iron by adding hexacyanoferrate deferasirox showed better effect on thallium elimination rate and improved functional state of liver and kidneys in patients with severe thallium salt poisoning.

Prussian blue nanoparticles: synthesis, surface modification, and biomedical applications.

Prussian blue nanoparticles (PBNPs) are a nanomaterial that presents unique properties and an excellent biocompatibility. They can be synthesized in mild conditions and can be derivatized with polymers and/or biomolecules. PBNPs are used in biomedicine as therapy and diagnostic agents. In biomedical imaging, PBNPs constitute contrast agents in photoacoustic and magnetic resonance imaging (MRI). They are a good adsorbent to be used as antidotes for poisoning with cesium and/or thallium ions. Moreover, the ability to convert energy into heat makes them useful photothermal agents (PAs) in photothermal therapy (PTT) or as nonantibiotic substances with antibacterial properties. Finally, PBNPs can be both reduced to Prussian white and oxidized to Prussian green. A large window of redox potential exists between reduction and oxidation, which result in the enzyme-like characteristics of these NPs.

Zinc-doped Prussian blue enhances photothermal clearance of Staphylococcus aureus and promotes tissue repair in infected wounds.

The application of photothermal therapy to treat bacterial infections remains a challenge, as the high temperatures required for bacterial elimination can damage healthy tissues. Here, we develop an exogenous antibacterial agent consisting of zinc-doped Prussian blue (ZnPB) that kills methicillin-resistant Staphylococcus aureus in vitro and in a rat model of cutaneous wound infection. Local heat triggered by the photothermal effect accelerates the release and penetration of ions into the bacteria, resulting in alteration of intracellular metabolic pathways and bacterial killing without systemic toxicity. ZnPB treatment leads to the upregulation of genes involved in tissue remodeling, promotes collagen deposition and enhances wound repair. The efficient photothermal conversion of ZnPB allows the use of relatively few doses and low laser flux, making the platform a potential alternative to current antibiotic therapies against bacterial wound infections.

Nanozyme-mediated catalytic nanotherapy for inflammatory bowel disease.

The overproduction of reactive oxygen species (ROS) is linked to inflammatory bowel disease (IBD) and causes oxidative damage to DNA, proteins, and lipids. These ROS promote the initiation and progression of ulcerative colitis (UC). This study proposes a unique concept of nanomaterials with intrinsic enzyme-like activity (nanozymes) to mediate catalytic nanotherapy for IBD. Methods: We first synthesized manganese Prussian blue nanozymes (MPBZs) with multi-enzyme activity. A dextran sulfate sodium (DSS)-induced mouse model of colitis was built. The ROS scavenging capacity and anti-inflammatory effects of the MPBZs were investigated. Results: As a proof of concept, MPBZs with multi-enzyme activity were constructed of variable valence elements (Mn and Fe) via a facile and efficient strategy. Due to the increased intestinal permeability and positively charged surfaces of inflamed mucosa in murine colitis, the prepared MPBZs with nanoscale sizes and negative charges preferentially accumulated at inflamed sites after oral administration. Importantly, MPBZs mediated catalytic nanotherapy for IBD in mice via a primary effect on the toll-like receptor signaling pathway without adverse side effects. Conclusion: MPBZs with multi-enzyme activity were constructed to treat IBD. This nanozyme-based approach is a promising strategy for catalytic nanotherapy in patients with colonic IBD.

Large-scale synthesis of monodisperse Prussian blue nanoparticles for cancer theranostics via an "in situ modification" strategy.

BACKGROUND: The intrinsic properties of Prussian blue (PB) nanoparticles make them an attractive tool in nanomedicine, including magnetic resonance imaging (MRI), photoacoustic imaging (PAI), and photothermal therapy (PTT) properties. However, there still remains the challenge of their poor dispersible stability in the physiological environment. In this study, we developed an efficient hydrothermal method to address the poor dispersible stability of PB nanoparticles in the physiological environment. MATERIALS AND METHODS: The concentration of H+, the mass of polyvinylpyrrolidone (PVP), and iron sources (K3[Fe(CN)6]) are very vital in the preparation of PB nanoparticles. Through exploring the preparation process, optimized PB nanoparticles (OPBs) with excellent physiological stability were prepared. Hydrodynamic diameter and UV-vis absorption properties were measured to verify the stability of the prepared OPBs. Properties of dual-mode imaging, including MRI/PAI, and PTT of OPBs were investigated both in vitro and in vivo. In addition, the in vivo biosafety of OPBs was systematically assessed. RESULTS: OPBs were stable in different environments including various media, pH, and temperatures for at least 90 days, indicating that they are suitable for biomedical application via intravenous administration and easily stored in a robust environment. Compared with other research into the synthesis of PB nanoparticles, the "in situ modification" synthesis of PB nanoparticles had advantages, including a simple process, low cost, and easy mass preparation. OPBs showed no significant signs of toxicity for 90 days. As a proof of concept, the OPBs served as dual-mode image contrast agents and photothermal conversion agents for cancer diagnosis and therapy both in vitro and in vivo. CONCLUSION: Our findings suggest a facile but efficient strategy with low cost to address the poor dispersible stability of PB nanoparticles in physiological environments. This will promote the development of further clinical transformations of PB nanoparticles, especially in cancer theranostics.

Prussian blue nanoparticles: Synthesis, surface modification, and application in cancer treatment.

This review outlines recently developed Prussian blue nanoparticle (PB NPs)-based multimodal imaging-guided chemo-photothermal strategies for cancer diagnosis and treatment in order to provide insight into the future of the field. The primary limitation of existing therapeutics is the lack of selectivity in drug delivery: they target healthy and cancerous cells alike. In this paper, we provide a thorough review of diverse synthetic and surface engineering techniques for PB NP fabrication. We have elucidated the various targeting approaches employed to deliver the therapeutic and imaging ligands into the tumor area, and outlined methods for enhancement of the tumor ablative ability of the NPS, including several important combinatorial approaches. In addition, we have summarized different in vitro and in vivo effects of PB NP-based therapies used to overcome both systemic and tumor-associated local barriers. An important new approach - PB NP-based immune drug delivery, which is an exciting and promising strategy to overcome cancer resistance and tumor recurrence - has been discussed. Finally, we have discussed the current understanding of the toxicological effects of PB NPs and PB NP-based therapeutics. We conclude that PB NP-based multimodal imaging-guided chemo-photothermal therapy offers new treatment strategies to overcome current hurdles in cancer diagnosis and treatment.

Ag-HPBs by a coating-etching strategy and their derived injectable implants for enhanced tumor photothermal treatment.

Herein, we demonstrate a coating-etching strategy to directly synthesize hollow Prussian blue (PB) nanocubes with well-dispersed Ag nanoparticles (denoted as Ag-HPB). The method is accomplished by introduction of PB precursors, K3Fe(CN)6 and Fe3+ into a reaction system containing AgNO3 and ascorbic acid, in which a series reactions contain formation of Ag nanoparticles, coating of PB on the nanoparticles, and diffusion of Ag into the PB frameworks occur. The strategy for preparation of the hollow structured Ag-HPB is intrinsically simple and does not require pre-preparation of any sacrificial templates or toxic etching agents. The obtained Ag-HPB nanocubes possess uniform size (69 nm), well-defined hollow structure, strong near-infrared photothermal conversion capacity, and excellent photoacoustic and magnetic resonance imaging abilities. Furthermore, an injectable photothermal implants are prepared for the first time by mixing the Ag-HPB nanocubes with clinically used biological glue, which significantly enhance photothermal anti-tumor efficacy, showing great potential for clinical tumor treatment.

Hyaluronic Acid Conjugated Magnetic Prussian Blue@Quantum Dot Nanoparticles for Cancer Theranostics.

A multifunctional nanotheranostic agent was developed by conjugating both hyaluronic acid and bovine serum albumin coated CuInS2-ZnS quantum dots onto the surface of magnetic Prussian blue nanoparticles. The obtained nanoagent could serve as an efficient contrast agent to simultaneously enhance near infrared (NIR) fluorescence and magnetic resonance (MR) imaging greatly. The coexistence of magnetic core and CD44 ligand hyaluronic acid was found to largely improve the specific uptake of the nanoagent by CD44 overexpressed HeLa cells upon applying an external magnetic field. Both NIR fluorescence and MR imaging in vivo proved high accumulation of the nanoagent at tumor site due to its excellent CD44 receptor/magnetic dual targeting capability. After intravenous injection of the nanoagent and treatment of external magnetic field, the tumor in nude mice was efficiently ablated upon NIR laser irradiation and the tumor growth inhibition was more than 89.95%. Such nanotheranostic agent is of crucial importance for accurately identifying the size and location of the tumor before therapy, monitoring the photothermal treatment procedure in real-time during therapy, assessing the effectiveness after therapy.

Toxicological evaluation of Prussian blue nanoparticles after short exposure of mice.

Prussian blue nanoparticle (PBNP), a new type of theranostic nanomaterial, had been used for cancer magnetic resonance imaging and photothermal therapy. However, their long-term toxicity after short exposure in vivo was still unclear. In this study, we investigated the dynamic changes of the biochemical and immunity indicators of mice after PBNPs injection through tail vein. Histological results showed that the PBNPs were mainly accumulated in liver and spleen. In the spleen, we found the frequency of T cells was starting to decrease after 1 day of PBNPs injection, but then slowly recovered to normal level after 60 days of injection. Meanwhile, the frequency of T cells in the blood was firstly decreased after the PBNPs injection, and then the T cell frequency kept increasing and recovered back to normal levels after 7 days of injection. The serum indexes of liver functions (alanine transaminase, aspartate transaminase, total bilirubin, and alkaline phosphatase) increased rapidly to a relatively high level only after 1 h of injection, which meant certain acute liver damage, but these indexes were gradually decreased to normal levels after 60 days of injection. These results indicate that PBNPs have acute toxicity in vivo, however, their long-term toxicity after short-time exposure is low, which might provide guidance for further applications of PBNPs in future.

[Reduction of 137caesium contamination in wild boars by supplementing offered food with ammonium-iron-hexa-cyanoferrate]. / Reduktion der 137Cäsium-Aktivität in Wildschweinen durch Zusatz von Ammonium-Eisen-Hexacyanoferrat ("Berliner Blau") zum Kirrfutter.

OBJECTIVE: This replication study investigated whether the 137caesium (137Cs) contamination of wild boars could be relevantly reduced under field conditions by adding ammonium-iron-hexa-cyanoferrate (AFCF; Prussian blue) to the food. MATERIAL AND METHODS: In 285 wild boars that had been shot in six Bavarian hunting territories during the season (November until May) between 01 November 2010 and 10 December 2011 137Cs contamination was analysed. Thirty-five animals originated from two hunting territories in which offered food had been supplemented with 1250 mg AFCF per kilogram food. RESULTS: The control animals showed a mean 137Cs contamination of 522 Bq/kg lean skeletal muscle meat. Direct (univariable) comparisons of the two experimental territories with the four control territories yielded a mean reduction in 137Cs contamination due to Prussian bluefeeding by -211 Bq/kg (p < 0.001). Multivariable models that took potential confounders into account (age, weight, sex, hunting date, territory) estimated the effect to be -344 Bq/kg (p < 0.05). CONCLUSION AND CLINICAL RELEVANCE: This replication study confirmed the finding of Kienzle et al. (12) who described a statistically significant reduction in 137Cs contamination by -380 Bq/kg due to the feeding of Prussian blue in other territories.

2,4-dinitrophenol partially alleviates ferrocyanide-induced toxicity in Drosophila melanogaster.

The toxicity of potassium ferrocyanide (PFC) and protective effects of 2,4-dinitrophenol (DNP) under PFC treatment were tested on the Drosophila melanogaster model system. Fly larvae were raised on food supplemented with PFC at concentrations of 1.0 mM and mixtures with DNP in concentrations of 0.50 and 1.25 mM, either alone or in combination with 1.0 mM PFC. Food supplementation with PFC decreased larvae viability or pupation height, whereas when larvae were fed by PFC and DNP combination the decrease was less pronounced. Larval exposure to PFC and mixtures of DNP and PFC lowered activities of aconitase. Larval treatment with PFC resulted in higher carbonyl protein, uric acid, and low molecular mass thiols content and higher activity of thioredoxin reductase in adult flies, while DNP in mixtures with PFC relieved these effects. Furthermore, treatment with PFC/DNP mixtures resulted in higher activities of superoxide dismutase and glutathione-S-transferase. It is proposed that PFC toxicity is mainly related to the cyanide and iron ions, released during its decomposition. The potential mechanisms of protective DNP effects against PFC toxicity are discussed.

New approach to the study of intraosseous vasculature.

BACKGROUND: The study of intraosseous vasculature has always been challenging due to the hard, calcified structure of bone. Techniques used to study intraosseous vasculature usually involve diaphanization with a Spalteholz technique, followed by X-ray imaging to display the radio contrast-injected blood vessels. These techniques produce results with fine detail when successfully executed. However, high failure rates and the extensive length of time required to perform these techniques are drawbacks. This paper describes a faster, alternative method for the study of intraosseous vasculature. METHOD: Five embalmed and two fresh shoulder girdles were harvested from six cadavers. Cannulas were inserted into the origins of the anterior (ACHA) and posterior (PCHA) circumflex humeral arteries and injected with ink diluted in water or 6% hydrogen peroxide. The arteries and their branches were dissected until they entered their respective bony foraminae. A hammer, chisel, bone nibbler and mounted needles were used to follow the intraosseous course of these arteries and their branches. RESULTS: The intraosseous vasculature was seen in all specimens. The branches of the main nutrient artery to the proximal humerus were followed until they reached articular cartilage or crossed cortical bone again to enter the rotator cuff tendons. DISCUSSION: An innovative, new approach to the study of intraosseous vasculature with direct visualization of the intraosseous arteries of the proximal humerus is described.

[Detoxification effects of two drugs in thallium -poisoned mice].

OBJECTIVE: To observe the thallium eliminating effect of prussian blue, pentetate zinc trisodium (Zn-DTPA), and their combined use in the treatment of acute thallium poisoning in mice. METHODS: Thallium poisoned mice were reproduced by oral administration of 0.2 ml thallous nitrate (3 mg/ml). They were assigned randomly to four groups according to the random number table method, namely, model group, prussian blue group, Zn-DTPA group and the combination therapy group, with 10 mice in each group. Prussian blue was administered orally [4.52 g×kg(-1)×d(-1), total four times], and Zn-DTPA was injected intraperitoneally [500 mg×kg(-1)×d(-1), one time]4 hours after giving thallium. The dosage of both drugs in combination treatment was as the same as described above. After treatment for 5 days, all the animals were sacrificed. Brain, intestine, kidney and liver of 1 mouse from each group were collected for pathological examination to observe the necrosis. Thallium contents of blood, brain, urine and feces from the other mice were determined. RESULTS: Pathological examination showed that the damage to intestine, kidney and liver was less obvious in treatment group compared with those of the model group. The effect was most obvious in the combination treatment group. However, brain damage was slightly improved. Thallium content in blood (mg/ml) of prussian blue group and the combination treatment group decreased obviously compared with the model group, and the decrease was more obvious in the combination treatment group (0.05 ± 0.01 vs. 0.18 ± 0.02). Thallium content in urine (mg/ml) and feces (mg/kg) was significantly increased after treatment, and the thallium elimination was most significant in the combined treatment group (urine: 11.34 ± 0.81 vs. 0.02 ± 0.01, feces: 13.11 ± 1.84 vs. 0.21 ± 0.07, both P < 0.01). Thallium content in brain was similar among all the groups. CONCLUSIONS: The single and combined use of prussian blue and Zn-DTPA could reduce the damage in intestine, kidney and liver. Combined use of prussian blue and Zn-DTPA for acute thallium poisoning mice is more efficacious than single use of either drug.

[Application of the ferrocin containing waste of wine-making for reduction of 137Cs tansit from forage to laboratory animal organisms and cow milk].

Addition to rat ration of ferrocin containing wastes of wine-making formed during the process of wine demetalization in the amount of 0.2 g per animal per day reduces the 137Cs content in organs and tissues in 1.5-7 times. Addition of the above-mentioned substance to the ration of milk cows in the amount of 10-16 g per day reduces the radionuclide content in milk 1.5-2 times in two weeks and more than 3 times in four weeks.

In vitro and in vivo evaluation of a novel ferrocyanide functionalized nanopourous silica decorporation agent for cesium in rats.

Novel decorporation agents are being developed to protect against radiological terrorist attacks. These sorbents, known as the self-assembled monolayer on mesoporous supports (SAMMS), are hybrid materials where differing organic moieties are grafted onto mesoporous silica (SiO(2)). In vitro experiments focused on the evaluation and optimization of SAMMS for capturing radiocesium ((137)Cs); therefore, based on these studies, a ferrocyanide copper (FC-Cu-EDA)-SAMMS was advanced for in vivo evaluation. In vivo experiments were conducted comparing the performance of the SAMMS vs. insoluble Prussian blue. Groups of jugular cannulated rats (4/treatment) were evaluated. Animals in Group I were administered (137)Cs chloride (approximately 40 microg kg(-1)) by intravenous (i.v.) injection or oral gavage; Group II animals were administered pre-bound (137)Cs-SAMMS or sequential Cs chloride + SAMMS (approximately 61 ng kg(-1)) by oral gavage; and Group III was orally administered (137)Cs chloride (approximately 61 ng kg(-1)) followed by either 0.1 g of SAMMS or Prussian blue. Following dosing, the rats were maintained in metabolism cages for 72 h and blood, urine, and fecal samples were collected for (137)Cs analysis (gamma counting). Rats were then humanely euthanized, and selected tissues analyzed. Orally administered (137)Cs chloride was rapidly and well absorbed (approximately 100% relative to i.v. dose), and the pharmacokinetics (blood, urine, feces, and tissues) were very comparable to the i.v. dose group. For both exposures the urine and feces accounted for 20 and 3% of the dose, respectively. The prebound (137)Cs-SAMMS was retained primarily within the feces (72% of the dose), with approximately 1.4% detected in the urine, suggesting that the (137)Cs remained tightly bound to SAMMS. SAMMS and Prussian blue both effectively captured available (137)Cs in the gut with feces accounting for 80-88% of the administered dose, while less than 2% was detected in the urine. This study suggests that the functionalized SAMMS outperforms Prussian blue in vitro at low pH, but demonstrates comparable in vivo sequestration efficacy at low exposure concentrations. The comparable response may be the result of the low (137)Cs chloride dose and high sorbent dosage that was utilized. Future studies are planned to optimize the performance of SAMMS in vivo over a broader range of doses and conditions.

Life-threatening Torsades de Pointes resulting from "natural" cancer treatment.

INTRODUCTION: Nonradioactive cesium chloride (CsCl) is used by some alternative medicine advocates as a treatment for cancer. The therapy was proven to be neither safe nor effective. Chronic use of CsCl has resulted in cases with severe cardiotoxicity. CASE REPORT: A 65-year-old lady presented to our hospital's accident and emergency department with recurrent syncope attacks. Electrocardiogram monitoring showed QT prolongation and transient Torsades de Pointes (TDP) ventricular tachycardia. She was taking anticancer naturopathic drugs for 6 weeks before admission. One of her naturopathic drugs was subsequently confirmed containing 89% CsCl by weight. Besides conventional treatment of QT prolongation and TDP, the patient was given a 4-week course of oral Prussian blue to enhance gastrointestinal elimination of cesium. The serum half-life of cesium was reduced from 61.7 to 29.4 days after the use of Prussian blue. QT prolongation was normalized in 27 days. DISCUSSION: To our knowledge, this is the first published case of nonradioactive cesium poisoning treated with Prussian blue. A transient rise in serum cesium level was observed during Prussian blue therapy. Possible explanations for this observation include poor drug compliance during outpatient treatment and redistribution of cesium from body stores. CONCLUSION: Nonradioactive CsCl poisoning can result in severe cardiotoxicity with QT prolongation and TDP ventricular tachycardia. The key points in the management of nonradioactive cesium poisoning include cessation of cesium exposure, vigorous electrolytes replacement, and oral Prussian blue therapy.