Systemic Toxicity Testing Research Articles

The antimicrobial peptide Microcin C7 inhibits the growth of Porphyromonas gingivalis and improves the perodontal status in a rat model.

This study aimed to investigate the antibacterial and anti-inflammatory effects of the antimicrobial peptide Microcin C7 for Porphyromonas gingivalis-associated diseases. Reverse-phase high-performance liquid chromatography revealed that Microcin C7 could remain 25.5% at 12 h in saliva. At a concentration of <10 mg ml-1, Microcin C7 showed better cytocompatibility, as revealed by a hemolysis test and a subchronic systemic toxicity test. Moreover, the minimum inhibitory concentration and minimum bactericidal concentration of Microcin C7 were analyzed using a broth microdilution method, bacterial growth curve, scanning electron microscopy, and confocal laser microscopy and determined to be 0.16 and 5 mg ml-1, respectively. Finally, in a rat model, 5 mg ml-1 Microcin C7 showed better performance in decreasing the expression of inflammatory factors (IL-1β, IL-6, IL-8, and TNF-α) and alveolar bone resorption than other concentrations. Microcin C7 demonstrated favorable biocompatibility, antibacterial activity, and anti-inflammatory effect, and could decrease the alveolar bone resorption in a rat model, indicating the promising potential for clinical translation and application on P. gingivalis-associated diseases.

Preparation of Ag@3D-TiO2 Scaffolds and Determination of its Antimicrobial Properties and Osteogenesis-promoting Ability.

The micro-nano structure of 3D-printed porous titanium (Ti) alloy with excellent performance in avoiding stress shielding and promoting bone tissue differentiation provides a new opportunity for the development of bone implants, but it necessitates higher requirements for bone tissue differentiation and the antibacterial properties of bone implants in clinical practice. This study investigated the preparation, antimicrobial properties, and osteogenesis-promoting ability of the 3D printed porous Ti alloy anodic oxidized Ag-carrying (Ag@3D-TiO2) scaffolds. The 3D printed porous Ti alloy (3D-Ti), anodized 3D printed porous Ti alloy (3D-TiO2), and Ag@3D-TiO2 scaffolds were synthesized using electron beam melting. The antimicrobial properties of the scaffolds were examined using antibacterial tests and their cytocompatibility was assessed using a cell proliferation assay and acridine orange/ethidium bromide (AO/EB) staining. In vitro cellular assays were used to investigate the effects of the scaffold microstructural features on cell activity, proliferation, and osteogenesis-related genes and proteins. In vivo animal experiments were used to evaluate the anti-inflammatory and osteogenesis-promoting abilities of the scaffolds. The Ag@3D-TiO2 scaffolds exhibited sustained anti-microbial activity over time, enhanced cell proliferation, facilitated osteogenic differentiation, and increased extracellular matrix mineralization. In addition, alkaline phosphatase (ALP), collagen type I (COL-I), and osteocalcin (OCN)-related genes and proteins were upregulated. In vivo animal implantation experiments, the anti-inflammatory effect of the Ag@3D-TiO2 scaffolds were observed using histology, and a large amount of fibrous connective tissue was present around it; the Ag@3D-TiO2 scaffolds were more bio-compatible with the surrounding tissues compared with 3D-Ti and 3D-TiO2; a large amount of uniformly distributed neoplastic bone tissue existed in their pores, and the chronic systemic toxicity test showed that the 3D-Ti, 3D-TiO2, and Ag@3D-TiO2 scaffolds are biologically safe. The goal of this study was to create a scaffold that exhibits antimicrobial properties and can aid bone growth, making it highly suitable for use in bone tissue engineering.

Cosmetic Formulations from Natural Sources: Safety Considerations and Legislative Frameworks in the European Union

Consumer preferences, safety, and sustainability aspects of conventional cosmetic ingredients have contributed to an increase in the demand for natural cosmetic ingredients and products. Naturally derived active cosmetic agents and excipients may come into contact with various naturally occurring and synthetic contaminants throughout the supply chain, and substantiating their safety is essential. This review examines the safety and legislative requirements applicable to natural cosmetic ingredients in the European Union (EU). Cosmetic safety requirements include technical data based on the ingredient profile, presence of hazards and the risks associated with the intended conditions of use. The hazard analysis includes screening for microbial contaminants such as aerobic mesophilic bacteria, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans; chemical contaminants such as lead, cadmium, arsenic, and mercury; and naturally occurring toxins, such as allergens. The toxicological assessment considers both local effects (such as skin sensitisation, eye/skin irritation, and photo-induced effects) and systemic effects (including acute dermal toxicity, sub-acute and sub-chronic toxicity, mutagenicity and carcinogenicity, reproductive toxicity, and toxicokinetics). The EU legislative requirements prohibit the use of animal-based tests for the toxicological evaluation of cosmetic ingredients, paving the way for alternatives termed as New Approach Methodologies (NAMs). The validation of NAMs is critical for their wider usage, and despite advancements, few have been validated, particularly for systemic toxicity testing. The use of NAMs in evaluating the safety of complex natural cosmetic ingredients is further examined.

Biosafety and chemical solubility studies of multiscale crystal-reinforced lithium disilicate glass-ceramics.

Lithium disilicate (Li2 Si2 O5 ) glass-ceramics are currently a more widely used all-ceramic restorative material due to their good mechanical properties and excellent aesthetic properties. However, they have a series of problems such as high brittleness and low fracture toughness, which has become the main bottleneck restricting its development. Therefore, in order to compensate for these shortcomings, we propose to prepare a reinforced glass-ceramics with better mechanical properties and to test the biosafety and chemical solubility of the material. Li2 Si2 O5 whiskers were synthesized by a one-step hydrothermal method, and multi-scale crystal-enhanced Li2 Si2 O5 glass-ceramics were prepared by reaction sintering. The biosafety of multi-scale crystal-reinforced Li2 Si2 O5 glass-ceramics was investigated by in vitro cytotoxicity test, rabbit pyrogen test, mice bone marrow micronucleus test, skin sensitization test, sub-chronic systemic toxicity test, and chronic systemic toxicity test. Additionally, the chemical solubility of multi-scale crystal-reinforced Li2 Si2 O5 glass-ceramics was investigated. The test results showed that the material was non-cytotoxic, non-thermogenic, non-mutagenic, non-sensitizing, and non-systemic. The chemical solubility, determined to be 377 ± 245 μg/cm2 , complied with the ISO 6872 standard for the maximum solubility of ceramic materials. Multi-scale crystal-reinforced Li2 Si2 O5 glass-ceramics' biosafety and chemical solubility met current normative criteria, and they can move on to mechanical property measurements (such as flexural strength test, fatigue life test, friction and wear property study, etc.) and bonding property optimization, which shows promise for future clinical applications.

3D printed polycaprolactone/β-tricalcium phosphate/carbon nanotube composite – Physical properties and biocompatibility

Three-dimensional (3D) printing is a bio-fabrication technique used to process tissue-engineered scaffolds for bone repair and remodeling. Polycaprolactone (PCL)/β-tricalcium phosphate (TCP) has been used as a base and osteoconductive biomaterial for bone tissue engineering in the past decades. The current study reveals the fabrication of a polycaprolactone (PCL)/β-tricalcium phosphate (TCP) scaffold by incorporating carbon nanotubes (CNT) via 3D printing. The physical properties and cytocompatibility of a new type of tissue engineering composite from polycaprolactone/β-tri-calcium phosphate/carbon nanotubes were investigated, and it was an absorbable scaffold prepared via furnace deposition 3D printing technology. The scaffold was designed with CAD software, and the composite material was fabricated via 3D printing. The printed composite material was tested for mechanical strength, scanning electron microscope (SEM) analysis, porosity calculation, systemic toxicity test, hemolysis rate determination, and effect on the proliferation of rat adipose-derived stem cells cultured in vitro. A composite scaffold with a length of 15 mm, width of 10 mm, and height of 5 mm was manufactured through CAD software drawing and 3D printing technology. Scanning electron microscopy measurements and analysis of the internal pore size of the stent are appropriate; the pores are interconnected, and the mechanical strength matches the strength of human cancellous bone. The calculated porosity of the stent was >60%, non-toxic, and non-hemolytic. The proliferation activity of the ADSC co-cultured with different scaffold materials was as follows: polycaprolactone/β-tricalcium phosphate/0.2% carbon nanotube scaffolds > polycaprolactone/β-tricalcium phosphate/0.1% carbon nanotube scaffolds > polycaprolactone/β-tricalcium phosphate/0.3% carbon nanotube scaffolds > polycaprolactone/β-tricalcium phosphate scaffolds (P < 0.05). The results showed that polycaprolactone/β-tricalcium phosphate/0.2% carbon nanotube scaffolds promoted the adhesion and proliferation of ADSC. The combination of 3D printing technology and CAD software can be used to print personalized composite stents, which have the characteristics of repeatability, high precision, and low cost. Through 3D printing technology, combining a variety of materials with each other can provide the greatest advantages of materials. The waste of resources was avoided. The prepared polycaprolactone/β-tri-calcium phosphate/0.2% carbon nanotube scaffold has a good pore structure and mechanical properties that mimic human cancellous bone, is non-toxic and non-hemolytic, and is effective in promoting ADSC proliferation in vitro. Given this correspondence, 3D printed scaffold shows good biocompatibility and strength, and the fabrication method provides a proof of concept for developing scaffolds for bone tissue engineering applications.

Towards the Clinical Translation of 3D PLGA/β-TCP/Mg Composite Scaffold for Cranial Bone Regeneration.

Recent years have witnessed the rapid development of 3D porous scaffolds with excellent biocompatibility, tunable porosity, and pore interconnectivity, sufficient mechanical strength, controlled biodegradability, and favorable osteogenesis for improved results in cranioplasty. However, clinical translation of these scaffolds has lagged far behind, mainly because of the absence of a series of biological evaluations. Herein, we designed and fabricated a composite 3D porous scaffold composed of poly (lactic-co-glycolic) acid (PLGA), β-tricalcium phosphate (β-TCP), and Mg using the low-temperature deposition manufacturing (LDM) technique. The LDM-engineered scaffolds possessed highly porous and interconnected microstructures with a porosity of 63%. Meanwhile, the scaffolds exhibited mechanical properties close to that of cancellous bone, as confirmed by the compression tests. It was also found that the original composition of scaffolds could be maintained throughout the fabrication process. Particularly, two important biologic evaluations designed for non-active medical devices, i.e., local effects after implantation and subchronic systemic toxicity tests, were conducted to evaluate the local and systemic toxicity of the scaffolds. Additionally, the scaffolds exhibited significant higher mRNA levels of osteogenic genes compared to control scaffolds, as confirmed by an in vitro osteogenic differentiation test of MC3T3-E1 cells. Finally, we demonstrated the improved cranial bone regeneration performance of the scaffolds in a rabbit model. We envision that our investigation could pave the way for translating the LDM-engineered composite scaffolds into clinical products for cranial bone regeneration.

Development of an AI-Assisted Embryo Selection System Using Iberian Ribbed Newts for Embryo-Fetal Development Toxicity Testing.

The 3Rs (Reduction, Refinement, Replacement) principle is driving the need for alternative methods in animal testing. Despite advancements in in vitro testing, complex systemic toxicity tests still necessitate in vivo approaches. The aim of this study was to develop a developmental toxicity test protocol using the Iberian ribbed newt (Pleurodeles waltl) as a model organism, integrating AI image analysis for embryo selection to improve test accuracy and reproducibility. We established a developmental toxicity test protocol based on the zebrafish test. Gonadotropin was administered to induce ovulation, and in vitro fertilization was performed. Embryos were imaged at 5-6 and 6-7 h post-fertilization. AI image analysis was utilized to assess embryo viability. The test chemical was administered 24-48 h post-fertilization, and morphological changes were observed daily until day 8. Additionally, a time-lapse photography system was constructed to monitor embryonic development. Out of 24 cultured embryos, 75% developed normally to the late tail bud stage or initial hatching stage, whereas 25% experienced developmental arrest or death. AI image analysis achieved high accuracy in classifying embryos, with overall accuracies of 92.0% and 92.9% for two learning models. The AI system demonstrated higher precision in the selection of viable embryos compared to visual inspection. The Iberian ribbed newt presents a viable alternative model for developmental toxicity testing, adhering to the 3Rs principles. The integration of AI image analysis substantially enhances the accuracy and reproducibility of embryo selection, providing a reliable method for evaluating developmental toxicity in pharmaceuticals.

Vaping product exposure system (VaPES): a novel in vitro aerosol deposition system

Objective Due to recent increases in the use of vaping devices, there is a high demand for research addressing the respiratory health effects of vaping products. Given the constantly changing nature of the vaping market with new devices, flavors, metals, and other chemicals rapidly emerging, there is a need for inexpensive and highly adaptable vaping device exposure systems. Here, we describe the design and validation of a novel in vitro aerosol exposure system for toxicity testing of vaping devices. Materials and methods We developed an inexpensive, open-source in vitro vaping device exposure system that produces even deposition, can be adapted for different vaping devices, and allows for experiments to be performed under physiological conditions. The system was then validated with deposition testing and a representative exposure with human bronchial epithelial cells (hBECs). Results The Vaping Product Exposure System (VaPES) produced sufficient and uniform deposition for dose-response studies and was precise enough to observe biological responses to vaping exposures. VaPES was adapted to work with both pod and cartridge-based vaping devices. Conclusion We have designed and validated a novel vaping device exposure system that will eliminate the need to use high-cost commercial exposure systems, lowering the barrier to entry of physiologically relevant vaping studies.

P14-31: Validation of an ALI exposure system for toxicity testing of particle emissions in outdoor environments

P14-31: Validation of an ALI exposure system for toxicity testing of particle emissions in outdoor environments

Natural mineral fibers: conducting inhalation toxicology studies – part A: Libby Amphibole aerosol generation and characterization method development

Background Asbestos has been classified as a human carcinogen, and exposure may increase the risk of diseases associated with impaired respiratory function. As the range of health effects and airborne concentrations that result in health effects across asbestos-related natural mineral fiber types are not fully understood, the National Institute of Environmental Health Sciences has established a series of research studies to characterize hazards of natural mineral fibers after inhalation exposure. This paper presents the method development work of this research project. Results A prototype nose-only exposure system was fabricated to explore the feasibility of generating natural mineral fiber aerosol for in vivo inhalation toxicity studies. The prototype system consisted of a slide bar aerosol generator, a distribution/delivery system and an exposure carousel. Characterization tests conducted using Libby Amphibole 2007 (LA 2007) demonstrated the prototype system delivered stable and controllable aerosol concentration to the exposure carousel. Transmission electron microscopy (TEM) analysis of aerosol samples collected at the exposure port showed the average fiber length and width were comparable to the bulk LA 2007. TEM coupled with energy dispersive spectrometry (EDS) and selected area electron diffraction (SAED) analysis further confirmed fibers from the aerosol samples were consistent with the bulk LA 2007 chemically and physically. Conclusions Characterization of the prototype system demonstrated feasibility of generating LA 2007 fiber aerosols appropriate for in vivo inhalation toxicity studies. The methods developed in this study are suitable to apply to a multiple-carousel exposure system for a rat inhalation toxicity testing using LA 2007.

A biocompatible silicon nitride dental implant material prepared by digital light processing technology

For decades, titanium has been the preferred material for dental implant fabrication. However, metallic ions and particles can cause hypersensitivity and aseptic loosening. The growing demand for metal-free dental restorations has also promoted the development of ceramic-based dental implants, such as silicon nitride. In this study, silicon nitride (Si3N4) dental implants were fabricated for biological engineering by photosensitive resin based digital light processing (DLP) technology, comparable to conventionally produced Si3N4 ceramics. The flexural strength was (770 ± 35) MPa by the three-point bending method, and the fracture toughness was (13.3 ± 1.1) MPa · m1/2 by the unilateral pre-cracked beam method. The elastic modulus measured by the bending method was (236 ± 10) GPa. To confirm whether the prepared Si3N4 ceramics possessed good biocompatibility, in vitro biological experiments were performed with the fibroblast cell line L-929, and preferable cell proliferation and apoptosis were observed at the initial stages. Hemolysis test, oral mucous membrane irritation test, and acute systemic toxicity test (oral route) further confirmed that the Si3N4 ceramics did not exhibit hemolysis reaction, oral mucosal stimulation, or systemic toxicity. The findings indicate that Si3N4 dental implant restorations with personalized structures prepared by DLP technology have good mechanical properties and biocompatibility, which has great application potential in the future.

Biaxial stretching of polytetrafluoroethylene in industrial scale to fabricate medical ePTFE membrane with node-fibril microstructure.

Expanded polytetrafluoroethylene (ePTFE) is promising in biomedical fields such as covered stents and plastic surgery owing to its excellent biocompatibility and mechanical properties. However, ePTFE material prepared by the traditional biaxial stretching process is with thicker middle and thinner sides due to the bowing effect, which poses a major problem in industrial-scale fabrication. To solve this problem, we design an olive-shaped winding roller to provide the middle part of the ePTFE tape with a greater longitudinal stretching amplitude than the two sides, so as to make up for the excessive longitudinal retraction tendency of the middle part when it is transversely stretched. The as-fabricated ePTFE membrane has, as designed, uniform thickness and node-fibril microstructure. In addition, we examine the effects of mass ratio of lubricant to PTFE powder, biaxial stretching ratio and sintering temperature on the performance of the resultant ePTFE membranes. Particularly, the relation between the internal microstructure of the ePTFE membrane and its mechanical properties is revealed. Besides stable mechanical properties, the sintered ePTFE membrane exhibits satisfactory biological properties. We make a series of biological assessments including in vitro hemolysis, coagulation, bacterial reverse mutation and in vivo thrombosis, intracutaneous reactivity test, pyrogen test and subchronic systemic toxicity test; all of the results meet the relevant international standards. The muscle implantation of the sintered ePTFE membrane into rabbits indicates acceptable inflammatory reactions of our sintered ePTFE membrane fabricated on industrial scale. Such a medical-grade raw material with the unique physical form and condensed-state microstructure is expected to afford an inert biomaterial potentially for stent-graft membrane.

Biosafety evaluation of BaSi2O2N2:Eu2+/PDMS composite elastomers.

In recent years, mechanoluminescent (ML) materials have shown great potential in stress sensing, mechanical energy collection and conversion, so they have attracted wide attention in the field of stomatology. In the early stage of this study, BaSi2O2N2:Eu2+ ML phosphors were synthesized by two-step high temperature solid state method, and then mixed with Polydimethylsiloxane (PDMS) in different proportions to obtain BaSi2O2N2:Eu2+/PDMS ML composites with different mass fractions (10%,20%,30%,40%,50%). Then its biosafety was evaluated by Cell Counting Kit-8 (CCK-8), Calcein-AM/PI fluorescence staining, hemolysis, oral mucosal irritation, acute and subacute systemic toxicity tests. The experimental results show that the biosafety of BaSi2O2N2:Eu2+/PDMS ML composite elastomers with different mass fraction is in line with the existing standards, and other related properties can be further studied.

Environmental Hormone Effects and Bioaccumulation of Propiconazole and Difenoconazole in Procypris merus.

Studying the bioaccumulation behavior and toxicity of triazole fungicides is a crucial part of comprehensively evaluating the environmental fate and aquatic toxicity.The current research aimed to reveal the toxic effects of propiconazole and difenoconazole on fish through acute toxicity test, bioaccumulation test and oxidase system activity determination. Here, the propiconazole and difenoconazole concentrations were 11.3mg/L and 31.2mg/L for LC50-96h, both having low toxicity. LC-MS/MS was used to determine the propiconazole and difenoconazole concentrations in five organs (muscle, gill, liver, intestine, and kidney) of Procypris meru. The findings indicate that the bioconcentration coefficients of propiconazole and difenoconazole in grass flower carp were 0.66-27.08 and 2.43-22.72, which belonged to medium enrichment pesticides. The bioconcentration coefficients decreased with the increase of exposure concentration. The two fungicides could induce oxidative stress in fish liver, and the activities of three antioxidant enzymes were inhibited in varying degrees (p < 0.05). The results showed that the content of T3 increased, and T4 decreased when exposed to one-tenth LC50 for 7days. This study shows that triazole pesticides have bioaccumulation risks on aquatic organisms and clear environmental hormonal effects.

The Survival Response of Earthworm (Eisenia fetida L.) to Individual and Binary Mixtures of Herbicides.

Frequent use of herbicides may impose a risk on non-target species. The objective was to test the combined toxic effect of binary herbicide mixtures—metribuzin:halosulfuron and metribuzin:flumioxazin—on non-target earthworms in two test systems: filter paper and a soil toxicity test system. The joint action experiments were independently run twice to substantiate the findings. The most potent individual herbicide was metribuzin, with a 50% lethal concentration (LC50) of 17.17 µg ai. cm−2 at 48 h in the filter paper test. The toxicity of the individual herbicides on the filter paper test was ranked as metribuzin>halosulfuron>flumioxazin. In the soil test, metribuzin and halosulfuron had high toxicity with an LC50 of 8.48 and 10.08 mg ai. kg−1, respectively, on day 14. Thus, the individual herbicide ranking did not change between the filter paper and artificial soil tests. The herbicide’s mixed effect in both test systems showed a consistent antagonistic effect relative to a Concentration Addition reference model. It indicates that the mixtures retracted the herbicide’s action in the earthworms.

First Food and Drug Administration Cleared Thin-Film Electrode for Intracranial Stimulation, Recording, and Monitoring of Brain Activity-Part 1: Biocompatibility Testing.

Subdural strip and grid invasive electroencephalography electrodes are routinely used for surgical evaluation of patients with drug-resistant epilepsy (DRE). Although these electrodes have been in the United States market for decades (first FDA clearance 1985), their fabrication, materials, and properties have hardly changed. Existing commercially available electrodes are made of silicone, are thick (>0.5 mm), and do not optimally conform to brain convolutions. New thin-film polyimide electrodes (0.08 mm) have been manufactured to address these issues. While different thin-film electrodes are available for research use, to date, only one electrode is cleared by Food and Drug Administration (FDA) for use in clinical practice. This study describes the biocompatibility tests that led to this clearance. Biocompatibility was tested using standard methods according to International Organization for Standardization (ISO) 10993. Electrodes and appropriate control materials were bent, folded, and placed in the appropriate extraction vehicles, or implanted. The extracts were used for in vitro and in vivo tests, to assess the effects of any potential extractable and leachable materials that may be toxic to the body. In vitro studies included cytotoxicity tested in L929 cell line, genotoxicity tested using mouse lymphoma assay (MLA) and Ames assay, and hemolysis tested in rabbit whole blood samples. The results indicated that the electrodes were non-cytotoxic, non-mutagenic, non-clastogenic, and non-hemolytic. In vivo studies included sensitization tested in guinea pigs, irritation tested in rabbits, acute systemic toxicity testing in mice, pyrogenicity tested in rabbits, and a prolonged 28-day subdural implant in sheep. The results indicated that the electrodes induced no sensitization and irritation, no weight loss, and no temperature increase. Histological examination of the sheep brain tissue showed no or minimal immune cell accumulation, necrosis, neovascularization, fibrosis, and astrocyte infiltration, with no differences from the control material. In summary, biocompatibility studies indicated that these new thin-film electrodes are appropriate for human use. As a result, the electrodes were cleared by the FDA for use in clinical practice [510(k) K192764], making it the first thin-film subdural electrode to progress from research to clinic. Its readiness as a commercial product ensures availability to all patients undergoing surgical evaluation for DRE.

Uptake and depuration of water-soluble fractions (WSF) of crude oil by the bivalve Corbicula fluminea (Müller) from Shatt Al-Arab river

This study includes laboratory uptake and depuration experiments. The bivalve C. fluminea is exposed to 5ml of WSF of Nahran–Umar crude oil for each liter of river water for 48 hours in the static toxicity test system. The exposure water and animals are periodically taken for analysis of total hydrocarbons concentrations by spectroflurometer. After the exposure period is over, the bivalve transferred to clean river water for depuration period of 48 hours. The experiments show that there are no mortalities due to WSF of Nahran–Umar crude oil. The bivalve is usually active during the 12–16 hours of the experiments. Hydrocarbons concentrations in the exposure water decreased are slowly in a linear fashion so that no measurable concentrations remained after 32 hours. Hydrocarbons accumulated by the bivalve are slowly during the first few hours of exposure. After 2 hours of exposure, the concentration of hydrocarbons is approximately 0.38 ppm in water, while in tissues it is 4.8 ppm. After 20 hours, hydrocarbons are present in water at a concentration of 0.09ppm, while the bivalve tissues contained 17.7 ppm hydrocarbons. After 25 hours of exposure, the hydrocarbons concentration decreased in the bivalve tissues. At the end of the exposure period, the bivalve contains approximately 7.4 ppm. After 48 hours in clean river water, tissues burden decreased to 0.22 ppm of hydrocarbons. The bivalve remained contaminated with small concentrations of hydrocarbons for 20 days. The complete depuration occurred at 26 days.&#x0D;

Application of the Adverse Outcome Pathway Concept to In Vitro Nephrotoxicity Assessment: Kidney Injury due to Receptor-Mediated Endocytosis and Lysosomal Overload as a Case Study.

Application of adverse outcome pathways (AOP) and integration of quantitative in vitro to in vivo extrapolation (QIVIVE) may support the paradigm shift in toxicity testing to move from apical endpoints in test animals to more mechanism-based in vitro assays. Here, we developed an AOP of proximal tubule injury linking a molecular initiating event (MIE) to a cascade of key events (KEs) leading to lysosomal overload and ultimately to cell death. This AOP was used as a case study to adopt the AOP concept for systemic toxicity testing and risk assessment based on in vitro data. In this AOP, nephrotoxicity is thought to result from receptor-mediated endocytosis (MIE) of the chemical stressor, disturbance of lysosomal function (KE1), and lysosomal disruption (KE2) associated with release of reactive oxygen species and cytotoxic lysosomal enzymes that induce cell death (KE3). Based on this mechanistic framework, in vitro readouts reflecting each KE were identified. Utilizing polymyxin antibiotics as chemical stressors for this AOP, the dose-response for each in vitro endpoint was recorded in proximal tubule cells from rat (NRK-52E) and human (RPTEC/TERT1) in order to (1) experimentally support the sequence of key events (KEs), to (2) establish quantitative relationships between KEs as a basis for prediction of downstream KEs based on in vitro data reflecting early KEs and to (3) derive suitable in vitro points of departure for human risk assessment. Time-resolved analysis was used to support the temporal sequence of events within this AOP. Quantitative response-response relationships between KEs established from in vitro data on polymyxin B were successfully used to predict in vitro toxicity of other polymyxin derivatives. Finally, a physiologically based kinetic (PBK) model was utilized to transform in vitro effect concentrations to a human equivalent dose for polymyxin B. The predicted in vivo effective doses were in the range of therapeutic doses known to be associated with a risk for nephrotoxicity. Taken together, these data provide proof-of-concept for the feasibility of in vitro based risk assessment through integration of mechanistic endpoints and reverse toxicokinetic modelling.

Nutritional and biological value of natural-bio shoots mung bean “Mungoltin”. Food and biological values

Based on the results of our own research, examination of scientific dossier materials and reference literature data, it was established that dry powder Mungoltin made from the shoots of mung beans produced by Oriona-Scorpion LLC (Uzbekistan) contains a sufficient amount of protein, minerals, vitamins and dietary fiber, does not have a negative impact on the health status of experimental animals and does not result in functional and material cumulation. Acute systemic toxicity testing with intragastric administration of Mungoltin was carried out in 18 adult white male rats. Animals were divided into 4 groups. The animals of the first group received a nutrition dose of 5000 mg/ kg; the white rats of the second group were administered with a dose of 7500 mg/kg, and the rodents in the third group were given a dose of 10000 mg/kg, respectively. The animals in group 4 (the reference group) received distilled water. Upon a prolonged intragastric exposure to Mungoltin, no changes in biochemical parameters were detected. The activity indicators of alkaline phosphatase, trans-aminase enzymes and total protein in the blood serum did not differ significantly from those found in the reference group. Therefore, using Mungoltin will not cause a cytotoxic effect in relation to normal highly proliferating cells in an organism. The results of histomorphological studies of tissues of internal organs upon intragastric administration of Mungoltin within 30 days confirm the absence of toxic effects. According to toxicity parameters under the conditions of the above acute experiments, Mungoltin can be attributed to class 5 practically as a non-toxic substance.

Biosafety evaluation of Li2Si2O5 whisker-reinforced glass-ceramics

Lithium disilicate (Li2Si2O5) glass-ceramic is a commonly used dental ceramic material. In this study, Li2Si2O5 whiskers were prepared by the hydrothermal method, mixed with Li2Si2O5 glass powders, and Li2Si2O5 whisker-reinforced glass-ceramics were prepared by reaction sintering. The biosafety of the new Li2Si2O5 glass-ceramics were evaluated by in vitro cytotoxicity, hemolysis, oral mucosal irritation, acute systemic toxicity, and subacute systemic toxicity (oral route) tests according to ISO 7405/ISO 10993 standards. The cytotoxicity test results showed that the cell growth of the experimental group was good, and the cell number and morphology were not significantly different from those of the blank group (P> 0.05). The toxicity grading for both experimental and blank control groups were 0. The hemolysis rate of the material was 1.25%, which indicated that it did not cause hemolytic reaction. The material was non-irritating to the oral mucosa. In acute systemic toxicity test, animals in the experimental group showed increased body weight, moved freely, with no signs of poisoning. The food utilization rate and relative growth rate (change of the weight) of rats in the subacute systemic toxicity test were not statistically different from those of the control group (P > 0.05). Preliminary evaluation of the biosafety of the Li2Si2O5 whisker-reinforced glass-ceramics showed that it met the existing regulatory standards, and further biosafety experiments can be conducted, following which the material may be expected to be applied in clinical practice.