RESUMO
Microplastics pollution and salinity intrusion in freshwater ecosystem is one of the worldwide climate change consequences those have negative impacts on the physiology of aquatic organisms. Hence, a 15-day experiment was carried out where Nile tilapia (Oreochromis niloticus) was exposed to different salinity gradients i.e. 0â¯, 3â¯, 6â¯, 9â¯, and 12⯠alone and along with 10â¯mg/L polyamide microplastics (PA-MP) in order to measure its effects on the hematology, gill, and intestinal morphology. The results exhibited that all the fish treated with PA-MP ingested microplastics and the quantity of accumulation was significantly greater in higher salinity gradients (9⯠and 12â¯). In addition, the PA-MP treated fish showed increased glucose level and at the same time reduced hemoglobin concentration with the increase of salinity. The percentages of abnormalities in erythrocytes both cellular (twin, teardrop and spindle shaped) and nuclear (notched nuclei, nuclear bridge and karyopyknosis) significantly enhanced with PA-MP exposure again in higher salinity treatments (9⯠and 12â¯). The principal component analysis (PCA) exhibited that the addition of 10â¯mg/L PA-MP negatively affected the hematology of Nile tilapia than that of salinity treatments alone. Besides, the exposure of PA-MP in 9⯠and 12⯠salinity gradients escalated the severity of histological damages in gills and intestine. Overall, this experiment affirms that the increase of salinity enhanced the microplastics ingestion and toxicity in Nile tilapia, therefore, PA-MP possibly is addressed as additional physiological stressors along with increased salinity gradients in environment.
Assuntos
Ciclídeos , Brânquias , Microplásticos , Nylons , Salinidade , Poluentes Químicos da Água , Animais , Microplásticos/toxicidade , Poluentes Químicos da Água/toxicidade , Ciclídeos/metabolismo , Brânquias/efeitos dos fármacos , Brânquias/patologia , Brânquias/metabolismo , Nylons/toxicidade , Intestinos/efeitos dos fármacosRESUMO
BACKGROUND: Exposure to micro- and nanoplastic particles (MNPs) in humans is being identified in both the indoor and outdoor environment. Detection of these materials in the air has made inhalation exposure to MNPs a major cause for concern. One type of plastic polymer found in indoor and outdoor settings is polyamide, often referred to as nylon. Inhalation of combustion-derived, metallic, and carbonaceous aerosols generate pulmonary inflammation, cardiovascular dysfunction, and systemic inflammation. Additionally, due to the additives present in plastics, MNPs may act as endocrine disruptors. Currently there is limited knowledge on potential health effects caused by polyamide or general MNP inhalation. OBJECTIVE: The purpose of this study is to assess the toxicological consequences of a single inhalation exposure of female rats to polyamide MNP during estrus by means of aerosolization of MNP. METHODS: Bulk polyamide powder (i.e., nylon) served as a representative MNP. Polyamide aerosolization was characterized using particle sizers, cascade impactors, and aerosol samplers. Multiple-Path Particle Dosimetry (MPPD) modeling was used to evaluate pulmonary deposition of MNPs. Pulmonary inflammation was assessed by bronchoalveolar lavage (BAL) cell content and H&E-stained tissue sections. Mean arterial pressure (MAP), wire myography of the aorta and uterine artery, and pressure myography of the radial artery was used to assess cardiovascular function. Systemic inflammation and endocrine disruption were quantified by measurement of proinflammatory cytokines and reproductive hormones. RESULTS: Our aerosolization exposure platform was found to generate particles within the micro- and nano-size ranges (thereby constituting MNPs). Inhaled particles were predicted to deposit in all regions of the lung; no overt pulmonary inflammation was observed. Conversely, increased blood pressure and impaired dilation in the uterine vasculature was noted while aortic vascular reactivity was unaffected. Inhalation of MNPs resulted in systemic inflammation as measured by increased plasma levels of IL-6. Decreased levels of 17ß-estradiol were also observed suggesting that MNPs have endocrine disrupting activity. CONCLUSIONS: These data demonstrate aerosolization of MNPs in our inhalation exposure platform. Inhaled MNP aerosols were found to alter inflammatory, cardiovascular, and endocrine activity. These novel findings will contribute to a better understanding of inhaled plastic particle toxicity.
Assuntos
Nylons , Pneumonia , Humanos , Ratos , Feminino , Animais , Ratos Sprague-Dawley , Nylons/toxicidade , Microplásticos , Exposição por Inalação/efeitos adversos , Exposição por Inalação/análise , Dilatação , Aerossóis e Gotículas Respiratórios , Pneumonia/induzido quimicamente , Pulmão , Inflamação/induzido quimicamente , Tamanho da Partícula , Líquido da Lavagem BroncoalveolarRESUMO
Nylon has been widely used all over the world, and most of it eventually enters the aquatic environment in the form of microplastics (MPs). However, the impact of Nylon MPs on aquatic ecosystem remains largely unknown. Thus, the long-term biological effects and toxicity mechanism of Nylon MPs on Microcystis aeruginosa (M. aeruginosa) were explored in this study. Results demonstrated that Nylon MPs had a dose-dependent growth inhibition of M. aeruginosa at the initial stage, and the maximum inhibition rate reached to 47.62% at the concentration of 100 mg/L. Meanwhile, Nylon MPs could obstruct photosynthesis electron transfer, reduce phycobiliproteins synthesis, destroy algal cell membrane, enhance the release of extracellular polymeric substances, and induce oxidative stress. Furthermore, transcriptomic analysis indicated that Nylon MPs dysregulated the expression of genes involved in tricarboxylic acid cycle, photosynthesis, photosynthesis-antenna proteins, oxidative phosphorylation, carbon fixation in photosynthetic organisms, and porphyrin and chlorophyll metabolism. According to the results of transcriptomic and biochemical analysis, the growth inhibition of M. aeruginosa is inferred to be regulated by three pathways: photosynthesis, oxidative stress, and energy metabolism. Our findings provide new insights into the toxicity mechanism of Nylon MPs on freshwater microalgae and valuable data for risk assessment of MPs.
Assuntos
Microcystis , Poluentes Químicos da Água , Ecossistema , Transporte de Elétrons , Microplásticos , Nylons/toxicidade , Estresse Oxidativo , Fotossíntese , Plásticos , Poluentes Químicos da Água/toxicidadeRESUMO
The safety of microplastics (MPs) and associated health effects has been one of the major concerns worldwide. However, the role of photoaging toward the risk of MPs in water ecosystems remains inconclusive yet. In this study, the size of polyamide (PA, â¼32.50 µm) MPs was obviously decreased after photoaging in water containing fulvic acid (FA) and humic acid (HA) (â¼19.75 and â¼24.30 µm, respectively). Nanoplastics were formed (4.65% and 2.03%, respectively) and hydrophilia and colloidal stability was improved due to the formation of oxygen-containing functional groups. FA-aged PA exhibited higher inhibition on body length and weight of developing zebrafish than HA-aged and pristine PA. Photoaged MPs in intestine were more difficult to be depurated by zebrafish, leading to the disappearance of intestinal folding, shedding of more enterocytes, and emaciation of intestinal microvilli. Dietary lipid digestion in larvae was inhibited by aged PA due to oxidative stress-triggered lipid peroxidation and inhibition of lipase activities and bile acids secretion. Exposure of photoaged MPs down-regulated genes (cd36, dgat1a, dgat2, mttp, etc.) associated with triglyceride resynthesis and transportation, resulting in lipid maladsorption and growth inhibition. Our findings highlight the potential negative effects of environmentally aged MPs on diet digestion and nutrient assimilation in fish.
Assuntos
Envelhecimento da Pele , Poluentes Químicos da Água , Animais , Ecossistema , Intestinos/química , Lipídeos , Microplásticos , Nylons/toxicidade , Plásticos , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidade , Peixe-ZebraRESUMO
The relevance of the environmental hazard evaluation of virgin plastics particles is problematic, as plastics almost never occur in a virgin state after being discarded into the environment. However, the producers or importers must evaluate the environmental effect of their products as they are produced. Many plastic types e.g., polyamide, polyethylene are already under pre-registration, according to the database of the European Chemicals Agency (ECHA), in order to restrict the placing on the market of polymers (as defined by Article 3(5) of EU's REACH regulation (Registration, Evaluation, Authorization & Restriction of Chemicals), as a substance or in a mixture (ECHA, 2019). However, the hazard of microplastics could not be evaluated without relevant data on its (eco)toxic effects. In this work, the long-term toxicity of virgin polyamide microplastic (PA-MP) (size from 0 to 180 µm) and UV-weathered virgin PA-MP was investigated in the controlled life cycle experiments conducted in accordance with the OECD guidelines for testing of chemicals using Chironomus riparius (OECD test 218). In addition, a three-generation test was conducted to understand the trans-generational toxicity potential of virgin PA-MP. After UV irradiation (26 d) the buoyancy and color of the particles was changed and the share of smaller particles (of a few micrometer size range) increased. The exposure of C. riparius larvae to UV-weathered PA-MP (1000 mg kg-1) during their life cycle (28 d), negatively affected their development and subsequent emergence as adults. However, the exposure to virgin PA-MP throughout the life cycle and also over three consecutive generations did not significantly reduced the number of emerged adults. From the point of view of environmental hazard, the virgin polyamide plastics have probably no long-term hazard to chironomids. While it may not be relevant as environmental pollutant in the strict sense, UV-weathering may turn it hazardous.
Assuntos
Chironomidae , Poluentes Químicos da Água , Animais , Microplásticos , Nylons/toxicidade , Plásticos/toxicidade , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidadeRESUMO
Microplastics (MPs) are ubiquitous in the environment and pose substantial threats to the water ecosystem. However, the impact of natural aging of MPs on their toxicity has rarely been considered. This study found that visible light irradiation with hydrogen peroxide at environmentally relevant concentration for 90 days significantly altered the physicochemical properties and mitigated the toxicity of polyamide (PA) fragments to infantile zebrafish. The size of PA particles was reduced from â¼8.13 to â¼6.37 µm, and nanoparticles were produced with a maximum yield of 5.03%. The end amino groups were volatilized, and abundant oxygen-containing groups (e.g., hydroxyl and carboxyl) and carbon-centered free radicals were generated, improving the hydrophilicity and colloidal stability of degraded MPs. Compared with pristine PA, the depuration of degraded MPs mediated by multixenobiotics resistance was much quicker, leading to markedly lower bioaccumulation in fish and weaker inhibition on musculoskeletal development. By integrating transcriptomics and transgenic zebrafish [Tg(lyz:EGFP)] tests, differences in macrophages-triggered proinflammatory effects, apoptosis via IL-17 signaling pathway, and antioxidant damages were identified as the underlying mechanisms for the attenuated toxicity of degraded MPs. This work highlights the importance of natural degradation on the toxicity of MPs, which has great implications for risk assessment of MPs.
Assuntos
Microplásticos , Poluentes Químicos da Água , Animais , Apoptose , Ecossistema , Larva , Macrófagos , Nylons/toxicidade , Estresse Oxidativo , Plásticos , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidade , Peixe-ZebraRESUMO
Microplastic polyamide 66 (PA66) was used to explore its mechanism of influence on the contaminants removal from aerobic granular sludge (AGS) and the corresponding change to the microbial community. Results showed that the removal pollution efficiency of the experimental groups with PA66 were inhibited during the early treatment stage. However, as the experiment progressed, the removal efficiencies of chemical oxygen demand (COD) (92.66%, 93.10%, 93.11%, 93.79%) and ammonia nitrogen (94.25%, 94.58%, 95.61%, 94.73%) were similar in the addition 0 g/L (A), 0.1 g/L (B), 0.2 g/L (C) and 0.5 g/L (D) PA66 beakers at the last 10 days. On the first day, the intensity of fluorescence peaks representing tryptophan protein-like and aromatic protein-like substances of loosely-bound extracellular polymeric substances (LB-EPS) indicated that the PA66 microplastic caused damage to the sludge structure, and the intensity of fluorescence peaks representing fulvic acid-like and humic acid-like substances were stronger than those in the control beaker (A). Microbial community analysis showed that the main phyla were Firmicutes (49.11%, 59.77%, 44.33%, 41.21%), Proteobacteria (26.44%, 11.96%, 31.44%, 19.4%) and Bacteroidetes (9.24%, 13.05%, 11.89%, 14.71%) in the four beakers. According to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, genes representing [T] Signal transduction mechanisms illustrated that adding PA66 microplastic resulted in more signaling molecules in the AGS.
Assuntos
Microbiota/efeitos dos fármacos , Microplásticos/toxicidade , Nylons/toxicidade , Esgotos/microbiologia , Amônia/análise , Benzopiranos/análise , Análise da Demanda Biológica de Oxigênio , Substâncias Húmicas/análise , Nitrogênio/análise , Esgotos/química , Eliminação de Resíduos LíquidosRESUMO
Microplastics attract widespread attention, including for their potential to transport toxic chemicals in the form of plasticisers and associated hydrophobic organic chemicals, such as polybrominated diphenyl ethers (PBDEs). The aims of this study were to investigate how nylon (polyamide) microplastics may affect PBDE accumulation in snails, and the acute effects of nylon particles and PBDEs on survival, weight change and inherent microbiome diversity and community composition of the pond snail Lymnaea stagnalis. Snails were exposed for 96â¯h to BDEs-47, 99, 100 and 153 in the presence and absence of 1% w/w nylon microplastics in quartz sand sediment. No mortality was observed over the exposure period. Snails not exposed to microplastics lost significantly more weight compared to those exposed to microplastics. Increasing PBDE concentration in the sediment resulted in an increased PBDE body burden in the snails, however microplastics did not significantly influence total PBDE uptake. Based on individual congeners, uptake of BDE 47 by snails was significantly reduced in the presence of microplastics. The diversity and composition of the snail microbiome was not significantly altered by the presence of PBDEs nor by the microplastics, singly or combined. Significant effects on a few individual operational taxonomic units (OTUs) occurred when comparing the highest PBDE concentration with the control treatment, but in the absence of microplastics only. Overall within these acute experiments, only subtle effects on weight loss and slight microbiome alterations occurred. These results therefore highlight that L. stagnalis are resilient to acute exposures to microplastics and PBDEs, and that microplastics are unlikely to influence HOC accumulation or the microbiome of this species over short timescales.
Assuntos
Éteres Difenil Halogenados/metabolismo , Lymnaea/efeitos dos fármacos , Microbiota/efeitos dos fármacos , Microplásticos/toxicidade , Poluentes Químicos da Água/metabolismo , Animais , Carga Corporal (Radioterapia) , Exposição Ambiental/análise , Retardadores de Chama/análise , Retardadores de Chama/metabolismo , Retardadores de Chama/toxicidade , Éteres Difenil Halogenados/análise , Éteres Difenil Halogenados/toxicidade , Lymnaea/metabolismo , Lymnaea/microbiologia , Nylons/toxicidade , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidadeRESUMO
Due to the increasing need of new treatment options against bacterial lung infections, novel antimicrobial peptides (AMPs) are under development. Local bioavailability and less systemic exposure lead to the inhalation route of administration. Combining AMPs with nanocarriers (NCs) into nanosystems (NSs) might be a technique for improved results. An air-liquid interface (ALI) in vitro inhalation model was set up including a human alveolar lung cell line (A549) and an optimized exposure system (P.R.I.T.® ExpoCube®) to predict acute local lung toxicity. The approach including aerosol controls (cupper-II-sulfate and lactose) delivered lowest observable adverse effect levels (LOAELs). Different combinations of AMPs (AA139, M33) and NCs (polymeric nanoparticles (PNPs), micelles and liposomes) were tested under ALI and submerged in vitro conditions. Depending on the nature of AMP and NCs, packing of AMPs into NSs reduced the AMP-related toxicity. Large differences were found between the LOAELs determined by submerged or ALI testing with the ALI approach indicating higher sensitivity of the ALI model. Since aerosol droplet exposure is in vivo relevant, it is assumed that ALI based results represents the more significant source than submerged testing for in vivo prediction of local acute lung toxicity. In accordance with the current state-of-the-art view, this study shows that ALI in vitro inhalation models are promising tools to further develop in vitro methods in the field of inhalation toxicology.
Assuntos
Antibacterianos/toxicidade , Nanopartículas/toxicidade , Peptídeos/toxicidade , Células A549 , Aerossóis , Antibacterianos/administração & dosagem , Infecções Bacterianas/tratamento farmacológico , Sobrevivência Celular/efeitos dos fármacos , Humanos , Lipossomos , Pulmão/efeitos dos fármacos , Pneumopatias/tratamento farmacológico , Metacrilatos/administração & dosagem , Metacrilatos/toxicidade , Micelas , Nanopartículas/administração & dosagem , Nylons/toxicidade , Peptídeos/administração & dosagemRESUMO
Microplastics are emerging contaminants in the marine environment. They enter the ocean in a variety of sizes and shapes, with plastic microfiber being the prevalent form in seawater and in the guts of biota. Most of the laboratory experiments on microplastics has been performed with spheres, so knowledge on the interactions of microfibers and marine organisms is limited. In this study we examined the ingestion of microfibers by the sea anemone Aiptasia pallida using three different types of polymers: nylon, polyester and polypropylene. The polymers were offered to both symbiotic (with algal symbionts) and bleached (without algal symbionts) anemones. The polymers were introduced either alone or mixed with brine shrimp homogenate. We observed a higher percentage of nylon ingestion compared to the other polymers when plastic was offered in the absence of shrimp. In contrast, we observed over 80% of the anemones taking up all types of polymers when the plastics were offered in the presence of shrimp. Retention time differed significantly between symbiotic and bleached anemones with faster egestion in symbiotic anemones. Our results suggest that ingestion of microfibers by sea anemones is dependent both on the type of polymers and on the presence of chemical cues of prey in seawater. The decreased ability of bleached anemones to reject plastic microfiber indicates that the susceptibility of anthozoans to plastic pollution is exacerbated by previous exposure to other stressors. This is particularly concerning given that coral reef ecosystems are facing increases in the frequency and intensity of bleaching events due to ocean warming.
Assuntos
Artemia/metabolismo , Nanoestruturas/toxicidade , Plásticos/toxicidade , Anêmonas-do-Mar/fisiologia , Poluentes da Água/análise , Poluentes da Água/metabolismo , Animais , Recifes de Corais , Ecossistema , Nylons/toxicidade , Poliésteres/toxicidade , Polipropilenos/toxicidade , Poluição da Água/análiseRESUMO
OBJECTIVE: To compare the mechanical and biological properties of newly developed hybrid ceramics filled and unfilled polyamide 12 (PA 12) for craniofacial reconstruction via a fused deposition modelling (FDM) framework. METHODS: 15wt% of zirconia (ZrO2) as well as 30, 35, and 40wt% of beta-tricalcium phosphate (ß-TCP) were compounded with PA 12, followed by the fabrication of filament feedstocks using a single screw extruder. The fabricated filament feedstocks were used to print the impact specimens. The melt flow rate, tensile properties of fabricated filament feedstocks, and 3D printed impact properties of the specimens were assessed using melt flow indexer, universal testing machine, and Izod pendulum tester, respectively. The microstructure of selected filament feedstocks and broken impact specimens were analysed using a field emission scanning electron microscope and universal testing machine. Human periodontal ligament fibroblast cells (HPdLF) were used to evaluate the cytotoxicity of the materials by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromid) (MTT) assay. RESULTS: Hybrid ceramics filled PA 12 indicated sufficient flowability for FDM 3D printing. The tensile strength of hybrid ceramics filled PA 12 filament feedstocks slightly reduced as compared to unfilled PA 12. However, the tensile modulus and impact strength of hybrid ceramics filled PA 12 increased by 8%-31% and 98%-181%, respectively. A significant increase was also detected in the cell viability of the developed composites at concentrations of 12.5, 25, 50 and 100mg/ml. SIGNIFICANCE: The newly developed hybrid ceramics filled PA 12 filament feedstock with improved properties is suitable for an FDM-based 3D printer, which enables the creation of patient-specific craniofacial implant at a lower cost to serve low-income patients.
Assuntos
Cerâmica/química , Fibroblastos/efeitos dos fármacos , Prótese Maxilofacial , Nylons/química , Desenho de Prótese/métodos , Fosfatos de Cálcio/química , Fosfatos de Cálcio/toxicidade , Cerâmica/toxicidade , Humanos , Técnicas In Vitro , Nylons/toxicidade , Ligamento Periodontal/citologia , Impressão Tridimensional , Resistência à Tração , Zircônio/química , Zircônio/toxicidadeRESUMO
The clinical success of chimeric antigen receptor (CAR) T cell immunotherapy in treating multiple blood cancers has created a need for efficient methods of ex vivo gene delivery to primary human T cells for cell engineering. Here, we synthesize and evaluate a panel of cationic polymers for gene delivery to both cultured and primary human T cells. We show that a subset of comb- and sunflower-shaped pHEMA-g-pDMAEMA polymers can mediate transfection with efficiencies up to 50% in the Jurkat human T cell line with minimal concomitant toxicity (>90% viability). We then optimize primary human T cell transfection conditions including activation time, cell density, DNA dose, culture media, and cytokine treatment. We demonstrate transfection of both CD4+ and CD8+ primary human T cells with messenger RNA and plasmid DNA at efficiencies up to 25 and 18%, respectively, with similarly high viability.
Assuntos
DNA/administração & dosagem , Portadores de Fármacos/química , Metacrilatos/química , Nylons/química , Poli-Hidroxietil Metacrilato/química , RNA Mensageiro/administração & dosagem , Linfócitos T/metabolismo , Transfecção/métodos , Sobrevivência Celular/efeitos dos fármacos , DNA/genética , Portadores de Fármacos/metabolismo , Portadores de Fármacos/toxicidade , Humanos , Células Jurkat , Metacrilatos/metabolismo , Metacrilatos/toxicidade , Nylons/metabolismo , Nylons/toxicidade , Plasmídeos/administração & dosagem , Plasmídeos/genética , Poli-Hidroxietil Metacrilato/metabolismo , Poli-Hidroxietil Metacrilato/toxicidade , RNA Mensageiro/genética , Linfócitos T/efeitos dos fármacosRESUMO
PEGylated PAMAM-G4 dendrimers with substitution percentages of 50% and intermediate size PEG chains (0.55 and 2.0 kDa) were synthesized and evaluated as solubility enhancers and potential supramolecular carriers for the poorly soluble drug Silybin (SIL). Aqueous solubility profiles revealed that the PEGylated system with 2.0 kDa chains induced a five-fold solubility increase for SIL and the largest drug-loading capacity within the systems under study with an average complex stoichiometry of 71:1 according to the Higuchi-Connors formulation for multiple binding sites. The supramolecular interaction between SIL and PEGylated PAMAM-G4 dendrimers was confirmed by 2D-NOESY experiments, which evidenced the simultaneous complexation of the drug in both PAMAM-G4 branches and outermost PEG chains. In vitro release studies showed that 2.0 kDa PEG chains induced a more extended release time compared with 0.5 kDa PEG chains. This result was attributed to the enhancement of PEG assistance to SIL complexation in systems with longer PEG chains, which are able to self-penetrate into dendrimer cavities and cooperate in the stabilization of SIL complexes, thus delaying the release of SIL from the supramolecular host. These results are valuable for the future design and development of novel PAMAM-based systems for SIL complexation and delivery.
Assuntos
Antioxidantes/administração & dosagem , Dendrímeros/química , Portadores de Fármacos/química , Nylons/química , Polietilenoglicóis/química , Silimarina/administração & dosagem , Antioxidantes/química , Sobrevivência Celular/efeitos dos fármacos , Dendrímeros/toxicidade , Portadores de Fármacos/toxicidade , Liberação Controlada de Fármacos , Células HEK293 , Humanos , Nylons/toxicidade , Polietilenoglicóis/toxicidade , Silibina , Silimarina/química , SolubilidadeRESUMO
Antiapoptotic Bcl-2 protein's upregulated expression is a key reason for drug resistance leading to failure of chemotherapy. In this report, a series of biocompatible amphiphilic cationic poly[(R)-3-hydroxybutyrate] (PHB)-b-poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) copolymer, comprising hydrophobic PHB block and cationic PDMAEMA block, is designed to codeliver hydrophobic chemotherapeutic paclitaxel and Bcl-2 converting gene Nur77/ΔDBD with enhanced stability, due to the micelle formation by hydrophobic PHB segment. This copolymer shows less toxicity but similar gene transfection efficiency to polyethyenimine (25k). More importantly, this codelivery approach by PHB-PDMAEMA leads to increased drug resistant HepG2/Bcl-2 cancer cell death, by increased expression of Nur77 proteins in the Bcl-2 present intracellular mitochondria. This work signifies for the first time that cationic amphiphilic PHB-b-PDMAEMA copolymers can be utilized for the drug and gene codelivery to drug resistant cancer cells with high expression of antiapoptosis Bcl-2 protein and the positive results are encouraging for the further design of codelivery platforms for combating drug resistant cancer cells.
Assuntos
Sistemas de Liberação de Medicamentos , Resistencia a Medicamentos Antineoplásicos , Hidroxibutiratos/química , Metacrilatos/química , Neoplasias/tratamento farmacológico , Nylons/química , Paclitaxel/uso terapêutico , Poliésteres/química , Proteínas Proto-Oncogênicas c-bcl-2/genética , Tensoativos/química , Cátions/química , DNA/metabolismo , Células HEK293 , Células Hep G2 , Humanos , Interações Hidrofóbicas e Hidrofílicas , Hidroxibutiratos/síntese química , Hidroxibutiratos/toxicidade , Metacrilatos/síntese química , Metacrilatos/toxicidade , Neoplasias/patologia , Nylons/síntese química , Nylons/toxicidade , Paclitaxel/farmacologia , Plasmídeos/metabolismo , Poliésteres/síntese química , Poliésteres/toxicidade , Proibitinas , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , TransfecçãoRESUMO
The practical application of engineered nanomaterials or nanoparticles like polyamidoamine (PAMAM) dendrimers has been promoted in medical devices or industrial uses. The safety of PAMAM dendrimers needs to be assessed when used as a drug carrier to treat brain disease. However, the effects of PAMAM on the human nervous system remain unknown. In this study, human neural progenitor cells cultured as a 3D neurosphere model were used to study the effects of PAMAM dendrimers on the nervous system. Neurospheres were exposed to different G4-PAMAM dendrimers for 72 h at concentrations of 0.3, 1, 3, and 10 µg/ml. The biodistribution was investigated using fluorescence-labeled PAMAM dendrimers, and gene expression was evaluated using microarray analysis followed by pathway and network analysis. Results showed that PAMAM dendrimer nanoparticles can penetrate into neurospheres via superficial cells on them. PAMAM-NH2 but not PAMAM-SC can inhibit neurosphere growth. A reduced number of MAP2-positive cells in flare regions were inhibited after 10 days of differentiation, indicating an inhibitory effect of PAMAM-NH2 on cell proliferation and neuronal migration. A microarray assay showed 32 dendrimer toxicity-related genes, with network analysis showing 3 independent networks of the selected gene targets. Inducible immediate early gene early growth response gene 1 (Egr1), insulin-like growth factor-binding protein 3 (IGFBP3), tissue factor pathway inhibitor (TFPI2), and adrenomedullin (ADM) were the key genes in each network, and the expression of these genes was significantly down regulated. These findings suggest that exposure of neurospheres to PAMAM-NH2 dendrimers affects cell proliferation and migration through pathways regulated by Egr1, IGFBP3, TFPI2, and ADM.
Assuntos
Dendrímeros/toxicidade , Células-Tronco Neurais/efeitos dos fármacos , Nylons/toxicidade , Adrenomedulina/genética , Adrenomedulina/metabolismo , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Biologia Computacional , Bases de Dados Genéticas , Dendrímeros/metabolismo , Relação Dose-Resposta a Droga , Proteína 1 de Resposta de Crescimento Precoce/genética , Proteína 1 de Resposta de Crescimento Precoce/metabolismo , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica/efeitos dos fármacos , Redes Reguladoras de Genes/efeitos dos fármacos , Glicoproteínas/genética , Glicoproteínas/metabolismo , Humanos , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Nanopartículas , Células-Tronco Neurais/metabolismo , Neurogênese/efeitos dos fármacos , Nylons/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Esferoides Celulares , Fatores de Tempo , Distribuição Tecidual , Transcriptoma/efeitos dos fármacosRESUMO
Pyrrole-imidazole (Py-Im) polyamides are high affinity DNA-binding small molecules that can inhibit protein-DNA interactions. In VCaP cells, a human prostate cancer cell line overexpressing both AR and the TMPRSS2-ERG gene fusion, an androgen response element (ARE)-targeted Py-Im polyamide significantly downregulates AR driven gene expression. Polyamide exposure to VCaP cells reduced proliferation without causing DNA damage. Py-Im polyamide treatment also reduced tumor growth in a VCaP mouse xenograft model. In addition to the effects on AR regulated transcription, RNA-seq analysis revealed inhibition of topoisomerase-DNA binding as a potential mechanism that contributes to the antitumor effects of polyamides in cell culture and in xenografts. These studies support the therapeutic potential of Py-Im polyamides to target multiple aspects of transcriptional regulation in prostate cancers without genotoxic stress.
Assuntos
Nylons/toxicidade , Animais , Antineoplásicos/química , Antineoplásicos/uso terapêutico , Antineoplásicos/toxicidade , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , DNA/química , DNA/metabolismo , Dano ao DNA/efeitos dos fármacos , DNA Topoisomerases/química , DNA Topoisomerases/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Imidazóis/química , Masculino , Camundongos , Nylons/síntese química , Nylons/química , Proteínas de Fusão Oncogênica/metabolismo , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Ligação Proteica , Pirróis/química , Receptores Androgênicos/metabolismo , Análise de Sequência de RNA , Transplante HeterólogoRESUMO
Safe delivery systems that can not only encapsulate hydrophobic drug molecules, but also release them in response to specific triggers are important in several therapeutic and biomedical applications. In this paper, we have designed a nanogel based on molecules that are generally recognized as safe (GRAS). We have shown that the resultant polymeric nanogels exhibit responsive molecular release and also show high in vitro cellular viability on HEK 293T, HeLa, MCF 7, and A549 cell lines. The toxicity of these nanogels was further evaluated with a highly sensitive assay using mouse preimplantation embryo development, where blastocysts were formed after 4 days of in vitro culture, and live pups were born when morulae/early blastocysts were transferred to the uteri of surrogate recipients. Our results indicate that these nanogels are nontoxic during mammalian development and do not alter normal growth or early embryo success rate.
Assuntos
Blastocisto/efeitos dos fármacos , Nylons/química , Polietilenoglicóis/química , Polietilenoimina/química , Animais , Sobrevivência Celular/efeitos dos fármacos , Feminino , Células HEK293 , Células HeLa , Humanos , Interações Hidrofóbicas e Hidrofílicas , Células MCF-7 , Camundongos , Nanogéis , Nylons/toxicidade , Polietilenoglicóis/toxicidade , Polietilenoimina/toxicidadeRESUMO
The purpose of this study is to develop biodegradable cationic polyamides for non-viral gene delivery and elucidate their structural effects on gene transfection activity. To this end, a group of novel cationic polyamides were synthesized by polycondensation reaction between different di-p-nitrophenyl esters and tertiary amine-containing primary diamines. These linear polyamides have flexible alkylene group (ethylene or propylene), protonable amino group and bioreducible disulfide linkage in the polyamide main chain. The alkylene group and disulfide linkage in these polyamides have a distinct effect on their gene delivery properties including buffering capacity, gene binding ability and intracellular gene release profile. Those cationic polyamides containing disulfide linkage and 1,4-bis(3-aminopropyl)piperazine (BAP) residue exhibited high buffering capacity (endosomal escape ability), high gene binding ability, and intracellular gene release ability, thus inducing fast gene nucleus translocation and robust gene transfection in vitro against different cell lines and rat bone marrow mesenchymal stem cells. Moreover, the transfection efficiencies in vitro were comparable or higher than those of 25 kDa branched polyethylenimine and Lipofectamine 2000 transfection agent as positive controls. These cationic polyamides and their polyplexes were of low cytotoxicity when an optimal transfection efficacy was achieved. In vivo transfection tests showed that bioreducible BAP-based polyamides were applicable for intravenous gene delivery in a mouse model, leading to higher level of transgene expression in the liver as compared to 22 kDa linear polyethylenimine as a positive control. These cationic polyamides provide a useful platform to elucidate the relationship between chemical functionalities and gene transfection activity.
Assuntos
Cátions/química , Nylons/química , Transfecção/métodos , Ácidos/química , Animais , Morte Celular/efeitos dos fármacos , Linhagem Celular , Endocitose/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Humanos , Camundongos , Nylons/síntese química , Nylons/toxicidade , TransgenesRESUMO
For the first time, polyelectrolyte complex based on poly[(2-dimethylamino) ethyl methacrylate] (PDMAEMA) and chondroitin sulfate (CS) was prepared. The properties of novel material and precursors were investigated by WAXS, FTIR, TGA, SEM and DLS analysis. The PDMAEMA/CS PECs presented hydrophilic-hydrophobic transition at pHs 6.0, 7.0 and 8.0 whereas the non-complexed PDMAEMA showed such a transition at pH 8.0 and not at pHs 6.0 and 7.0. Studies of CS release from PECs at pHs 6 and 8 confirmed that the samples possess the potential to release the CS in alkaline and not in acidic conditions. Since PECs are thermo-responsive due to the reduction of LCST caused by the increase in pH, the release of CS was dependent on temperature and pH factors. Cytotoxicity assays using healthy VERO cells showed that the complexation between CS and PDMAEMA increased the PECs' biocompatibility related to PDMAEMA. However, the biocompatibility depends on the amount of CS present in the PECs.
Assuntos
Sulfatos de Condroitina/química , Composição de Medicamentos , Metacrilatos/química , Nylons/química , Animais , Sobrevivência Celular/efeitos dos fármacos , Chlorocebus aethiops , Sulfatos de Condroitina/toxicidade , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Eletrólitos , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Metacrilatos/toxicidade , Nylons/toxicidade , Solubilidade , Propriedades de Superfície , Temperatura , Células VeroRESUMO
Pyrrole-imidazole polyamides targeted to the androgen response element were cytotoxic in multiple cell lines, independent of intact androgen receptor signaling. Polyamide treatment induced accumulation of S-phase cells and of PCNA replication/repair foci. Activation of a cell cycle checkpoint response was evidenced by autophosphorylation of ATR, the S-phase checkpoint kinase, and by recruitment of ATR and the ATR activators RPA, 9-1-1, and Rad17 to chromatin. Surprisingly, ATR activation was accompanied by only a slight increase in single-stranded DNA, and the ATR targets RPA2 and Chk1, a cell cycle checkpoint kinase, were not phosphorylated. However, ATR activation resulted in phosphorylation of the replicative helicase subunit MCM2, an ATR effector. Polyamide treatment also induced accumulation of monoubiquitinated FANCD2, which is recruited to stalled replication forks and interacts transiently with phospho-MCM2. This suggests that polyamides induce replication stress that ATR can counteract independently of Chk1 and that the FA/BRCA pathway may also be involved in the response to polyamides. In biochemical assays, polyamides inhibit DNA helicases, providing a plausible mechanism for S-phase inhibition.