RESUMEN
Human Fortilin, an antiapoptotic protein, has also been implicated in several diseases; however, several potential uses of fortilin have also been proposed. Bearing the implications of fortilin in mind, fortilin analog, which has no complication with diseases, is required. Since a recombinant full-length fortilin from Fenneropenaeus merguiensis (rFm-Fortilin (FL)) reported only 44% (3e-27) homologous to human fortilin, therefore the biological activities of the Fm-Fortilin (FL) and its fragments (F2, F12, and F23) were investigated for potential use against HEMA toxicity from filling cement to pulp cell. The rFm-Fortilin FL, F2, 12, and F23 were expressed and assayed for proliferation activity. The rFm-Fortilin (FL) showed proliferation activity on human dental pulp cells (HDPCs) and protected the cells from 2-hydroxy-ethyl methacrylate (HEMA) at 1-20 ng/ml. In contrast, none of the rFm-Fortilin fragments promoted HDPC growth that may be due to a lack of three conserved amino acid residues together for binding with the surface of Rab GTPase for proliferative activity. In addition, rFm-Fortilin (FL) activated mineralization and trend to suppressed production of proinflammatory cytokines, including histamine (at 10 ng/ml) and TNF-α (at 100 ng/ml). Besides, the rFm-Fortilin (FL) did not mutate the Chinese hamster ovary (CHO) cell. Therefore, the rFm-Fortilin (FL) has the potential use as a supplementary medical material to promote cell proliferation in patients suffering severe tooth decay and other conditions.
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Proteínas de Artrópodos/farmacología , Penaeidae/química , Animales , Proteínas de Artrópodos/genética , Proteínas de Artrópodos/aislamiento & purificación , Células Cultivadas , Pulpa Dental/citología , Pulpa Dental/efectos de los fármacos , Histamina/metabolismo , Metacrilatos/toxicidad , Proteínas Recombinantes , Alineación de Secuencia , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
New insights about nanomaterials' biodistribution revealed their ability to achieve tumor accumulation by taking advantage from the dynamic vents occurring in tumor's vasculature. This paradigm-shift emphasizes the importance of extending nanomaterials' blood circulation time to enhance their tumor uptake. The classic strategy to improve nanomaterials' stability during circulation relies on their functionalization with poly(ethylene glycol). However, recent reports have been showing that PEGylated nanomaterials can suffer from the accelerated blood clearance phenomenon, emphasizing the importance of developing novel coatings for functionalizing the nanomaterials. To address this limitation, the modification of natural carriers' surface to enhance their stability appears to be a promising strategy. Herein, sulfobetaine methacrylate (SBMA)-functionalized bovine serum albumin (BSA) was synthesized for the first time to investigate the capacity of this modification to improve the resulting nanoparticles' physicochemical properties, colloidal stability and in vitro performance. This novel polymer was then employed in the formulation of nanoparticles loaded with IR780 for application in breast cancer phototherapy (IR/SBMA-BSA NPs). When compared to their non-functionalized equivalents, the IR/SBMA-BSA NPs presented a neutral surface charge and a higher stability in biologically relevant media. Due to these features, the IR/SBMA-BSA NPs could achieve a 1.9-fold greater uptake by breast cancer cells than IR/BSA NPs. Furthermore, the IR/SBMA-BSA NPs were cytocompatible towards normal cells and reduced breast cancer cells' viability up to 42%. The phototherapy mediated by IR/SBMA-BSA NPs could further decrease cancer cells' viability to about 12%. Overall, the IR/SBMA-BSA NPs have enhanced features that propel their application in breast cancer phototherapy.
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Neoplasias de la Mama/tratamiento farmacológico , Portadores de Fármacos , Indoles/farmacología , Metacrilatos/química , Nanopartículas , Fármacos Fotosensibilizantes/farmacología , Fototerapia , Albúmina Sérica Bovina/química , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Composición de Medicamentos , Estabilidad de Medicamentos , Femenino , Humanos , Indoles/química , Indoles/metabolismo , Células MCF-7 , Metacrilatos/toxicidad , Fármacos Fotosensibilizantes/química , Fármacos Fotosensibilizantes/metabolismo , Albúmina Sérica Bovina/toxicidad , Propiedades de SuperficieRESUMEN
Indomethacin is a non-steroidal anti-inflammatory agent included in one of the most commonly used drug classes worldwide. The use of this drug results in certain side effects, including gastrointestinal complications. Therefore, there exists a need to develop better methods for the delivery of such drugs into the body, such as those employing nanoparticles. The aim of the present study was to evaluate the cytotoxic and genotoxic effects of indomethacin-loaded Eudragit(®) L 100 nanocapsules (NI; based on methacrylic acid and methyl methacrylate) on cells unable (lymphocytes) and able to metabolize drugs (HepG2 cells), using comet and cytokinesis-block micronucleus (CBMN) assays in vitro. Cells were exposed to NI at concentrations of 5, 10, 50, 125, 250, and 500 µg/mL. The comet assay showed that NI induced no significant DNA damage in either cell type at any of the concentrations tested. The CBMN test confirmed these results; however, the highest concentration of 500 µg/mL resulted in a small but statistically significant clastogenic/aneugenic effect in HepG2 cells. These findings should encourage the development of new investigations of this nanomaterial as a delivery vehicle for anti-inflammatory drugs, such as indomethacin.
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Antiinflamatorios no Esteroideos/toxicidad , Excipientes/toxicidad , Indometacina/toxicidad , Metacrilatos/toxicidad , Nanocápsulas/toxicidad , Polímeros/toxicidad , Adolescente , Adulto , Supervivencia Celular/efectos de los fármacos , Daño del ADN , Evaluación Preclínica de Medicamentos , Femenino , Células Hep G2 , Humanos , Linfocitos/efectos de los fármacos , Linfocitos/fisiología , Masculino , Adulto JovenRESUMEN
A new generation of photothermal theranostic agents based on assembling 6 nm gold nanoparticles (AuNPs) is developed by using a novel comb-like amphipathic polymer as the template. The small AuNPs are assembled into DOX@gold nanomicelles, which show strong absorbance in the near-infrared region, for multimodal bioimaging and highly effective in vivo chemotherapy and photothermal therapy.
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Materiales Biocompatibles , Compuestos de Oro , Nanopartículas del Metal , Micelas , Polímeros , Adenocarcinoma/patología , Adenocarcinoma/terapia , Animales , Materiales Biocompatibles/uso terapéutico , Materiales Biocompatibles/toxicidad , Supervivencia Celular , Estudios de Factibilidad , Fibroblastos/fisiología , Compuestos de Oro/uso terapéutico , Compuestos de Oro/toxicidad , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Terapia por Láser/instrumentación , Células MCF-7 , Nanopartículas del Metal/uso terapéutico , Nanopartículas del Metal/toxicidad , Metacrilatos/toxicidad , Ratones , Imagen Multimodal/instrumentación , Trasplante de Neoplasias , Fototerapia/instrumentación , Polihidroxietil Metacrilato/toxicidad , Polímeros/uso terapéutico , Polímeros/toxicidad , Nanomedicina Teranóstica/instrumentación , Venas Umbilicales/citología , Venas Umbilicales/fisiologíaRESUMEN
Dental materials that can promote cell proliferation and function is required for regenerative pulp therapy. Resin modified glass ionomer cement (RMGIC), a broadly used liner or restorative material, can cause apoptosis to pulp cells mainly due to HEMA (2-hydroxyethyl methacrylate), the released residual monomer. Recent studies found that chitosan and albumin could promote release of protein in GIC while translationally controlled tumor protein (TCTP) has an anti-apoptotic activity against HEMA. The aim of this study was to examine the effect of chitosan and albumin modified RMGIC (Exp-RMGIC) supplemented with TCTP on pulp cell viability and mineralization. Exp-RMGIC+TCTP was composed of RMGIC powder incorporated with 15 % of chitosan, 5 % albumin and supplemented with TCTP mixed with the same liquid components of RMGIC. The effect of each specimen on pulp cells was examined using the Transwell plate. From the MTT assay, Exp-RMGIC+TCTP had the highest percentages of viable cells (P < 0.05) at both 24 and 74 h. Flow cytometry revealed that, after 24 h, Exp-RMGIC+TCTP gave the lowest percentages of apoptotic cells compared to other groups. There was no difference in alkaline phosphatase (ALP) activity among different formula of the specimens, while cells cultured in media with TCTP had higher ALP activity. Von Kossa staining revealed that RMGIC+TCTP, and Exp-RMGIC+TCTP had higher percentages of calcium deposit area compared to those without TCTP. It was concluded that Exp-RMGIC supplemented with TCTP had less cytotoxicity than RMGIC and can protect cells from apoptosis better than RMGIC supplemented with TCTP.
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Compuestos de Aluminio/química , Biomarcadores de Tumor/administración & dosificación , Quitosano/química , Pulpa Dental/citología , Pulpa Dental/efectos de los fármacos , Fluoruros/química , Cementos de Ionómero Vítreo/química , Compuestos de Silicona/química , Fosfatasa Alcalina/metabolismo , Animales , Apoptosis/efectos de los fármacos , Biomarcadores de Tumor/genética , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Pulpa Dental/fisiología , Cementos de Ionómero Vítreo/toxicidad , Humanos , Ensayo de Materiales , Metacrilatos/química , Metacrilatos/toxicidad , Penaeidae/genética , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/genética , Regeneración/efectos de los fármacos , Proteína Tumoral Controlada Traslacionalmente 1RESUMEN
Neutral Methacrylate Copolymer is a fully polymerised copolymer used in the pharmaceutical industry to permit pH-independent delayed release of active ingredients from oral dosage forms. This function has potential use with food supplements and this article describes available information on the safety of the substance. Oral administration of radiolabelled copolymer to rats resulted in the detection of chemically unchanged copolymer in the faeces, with negligible absorption. Safety studies revealed no adverse toxicity following repeated administration at doses of up to 2000 mg/kg bw/d in a sub-chronic study in rats or 250 mg/kg bw/d in a sub-chronic study in dogs. No reproductive toxicity occurred at up to 2000 mg/kg bw/d in rats or rabbits. The substance shows no evidence of genotoxicity, has low acute toxicity and no irritation or sensitisation potential. An ADI value of 20 mg/kg bw was concluded from two alternative approaches. Daily exposure from use in dietary supplements is estimated as up to 10.0 mg/kg bw in adults and 13.3 mg/kg bw in children. There would therefore appear to be no safety concerns under the intended conditions of use. The information provided is intended to support an evaluation that the substance may be "generally recognized as safe" (GRAS).
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Seguridad de Productos para el Consumidor , Excipientes/toxicidad , Aditivos Alimentarios/toxicidad , Metacrilatos/toxicidad , Animales , Evaluación Preclínica de Medicamentos , Excipientes/química , Aditivos Alimentarios/química , Aditivos Alimentarios/farmacocinética , Metacrilatos/química , Metacrilatos/farmacocinética , Microscopía Electrónica de Rastreo , Nivel sin Efectos Adversos Observados , Conejos , Ratas , Propiedades de Superficie , Pruebas de Toxicidad/métodosRESUMEN
A study of the in vitro sensitivity of 12 isolates of Phytophthora infestans to metalaxyl, azoxystrobin, dimethomorph, cymoxanil, zoxamide and mancozeb, was conducted. The isolates derived from infected potato leaves collected at eight different localities in Serbia during 2005-2007. The widest range of EC(50) values for mycelial growth of the isolates was recorded for metalaxyl. They varied from 0.3 to 3.9 µg mL(-1) and were higher than those expected in a susceptible population of P. infestans. The EC(50) values of the isolates were 0.16-0.30 µg mL(-1) for dimethomorph, 0.27-0.57 µg mL(-1) for cymoxanil, 0.0026-0.0049 µg mL(-1) for zoxamide and 2.9-5.0 µg mL(-1) for mancozeb. The results indicated that according to effective concentration (EC(50)) the 12 isolates of P. infestans were sensitive to azoxystrobin (0.019-0.074 µg mL(-1)), and intermediate resistant to metalaxyl, dimethomorph and cymoxanil. According to resistance factor, all P. infestans isolates were sensitive to dimethomorph, cymoxanil, mancozeb and zoxamide, 58.3% of isolates were sensitive to azoxystrobin and 50% to metalaxyl. Gout's scale indicated that 41.7% isolates were moderately sensitive to azoxystrobin and 50% to metalaxyl.
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Plaguicidas/toxicidad , Phytophthora infestans/efectos de los fármacos , Enfermedades de las Plantas/parasitología , Solanum tuberosum/parasitología , Acetamidas/toxicidad , Alanina/análogos & derivados , Alanina/toxicidad , Amidas/toxicidad , Maneb/toxicidad , Metacrilatos/toxicidad , Morfolinas/toxicidad , Phytophthora infestans/aislamiento & purificación , Pirimidinas/toxicidad , Serbia , Estrobilurinas , Zineb/toxicidadRESUMEN
PURPOSE: The aim of this study was to evaluate the ultrastructural characteristics and ionic profile of U937 cells after exposure to 2-hydroxyethyl methacrylate (HEMA) to shed light on the cytotoxicity of this dental adhesive and its relation to mechanisms of cell death. MATERIALS AND METHODS: U937 human monoblastic cells were incubated in RPMI 1640 culture medium and exposed to HEMA at LD50. Structural changes after 5, 15, 30, 60, and 120 min were observed with transmission electron microscopy. Ionic content of Na, K, Cl, Mg, P and S was evaluated by quantitative electron probe X-ray microanalysis. RESULTS: Our results in human monoblastic cell line U937 establish that exposure to HEMA at LD50 led to a singular mechanism of cell death characterized by changes in the morphology and ultrastructure of the cells (cell size, blebs, and organelle structure) compatible with apoptosis, but without changes in nuclear ultrastructure. These findings were consistent with our microanalytical data, which revealed a significant increase in intracellular Na and a decrease in K, along with a significant initial decrease in Cl concentration followed later (120 min) by an increase. CONCLUSION: All three lines of evidence (cell morphology, ultrastructural changes, and ionic profile) showed that HEMA at LD50 led to a hybrid process of cell death. We suggest that apoptosis and necrosis are part of a continuum comprising a single process of cell death.
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Materiales Dentales/toxicidad , Metacrilatos/toxicidad , Monocitos/efectos de los fármacos , Apoptosis/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Núcleo Celular/efectos de los fármacos , Núcleo Celular/ultraestructura , Tamaño de la Célula/efectos de los fármacos , Cloro/análisis , Microanálisis por Sonda Electrónica , Humanos , Dosificación Letal Mediana , Magnesio/análisis , Microscopía Electrónica de Transmisión , Monocitos/ultraestructura , Orgánulos/efectos de los fármacos , Orgánulos/ultraestructura , Fósforo/análisis , Potasio/análisis , Sodio/análisis , Azufre/análisis , Factores de Tiempo , Células U937RESUMEN
OBJECTIVES: To evaluate the cytotoxicity of Shellac F, a new fluoride varnish, and its effects on human dentin hydraulic conductance. METHODS: Shellac F was compared to another fluoride varnish (Duraphat) and a fluoride containing desensitizing agent (Isodan). The cytotoxicity test was performed on human gingival fibroblasts and through dentin slice on human pulp fibroblasts. The hydraulic conductance (Lp) was recorded by fluid filtration with a Flodec device under a constant pressure (15 cm H2O). The treated surface of the dentin disks and their sections were also investigated with SEM. RESULTS: The cytotoxicity test on gingival fibroblasts revealed that Duraphat was the least cytotoxic material, followed by Shellac F then Isodan. With dentin slice interposition, a lower level of cytotoxicity was obtained. All of them showed a lower cytotoxicity decreasing on further dilutions (p<0.001). The measurement of hydraulic conductance showed that all materials resulted in a significant decrease in dentin permeability after 24h comprising between 60 and 76%, but there was no statistically significant difference among the materials. This decrease was still over 50% of the initial values after 7 days for all three materials. SEM investigation showed dentin tubules covered with a thick layer of Shellac F or Duraphat whilst no material was observed on dentin surfaces treated with Isodan. SIGNIFICANCE: Shellac F showed an adequate cellular compatibility and a significant effect on human dentin hydraulic conductance. This indicates that the new material is safe and seems to be effective as a potential desensitizing agent.
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Sensibilidad de la Dentina/fisiopatología , Fluoruros Tópicos/farmacología , Fluoruro de Sodio/farmacología , Células Cultivadas , Pulpa Dental/efectos de los fármacos , Dentina/efectos de los fármacos , Dentina/ultraestructura , Permeabilidad de la Dentina/efectos de los fármacos , Evaluación Preclínica de Medicamentos , Fibroblastos/efectos de los fármacos , Fluoruros Tópicos/toxicidad , Encía/efectos de los fármacos , Humanos , Ensayo de Materiales , Metacrilatos/farmacología , Metacrilatos/toxicidad , Microscopía Electrónica de Rastreo , Nitratos/farmacología , Nitratos/toxicidad , Presión , Fluoruro de Sodio/toxicidad , Factores de TiempoRESUMEN
OBJECTIVES: The aim of this study was to examine the relationship between the monomers eluted from dentin-bonding systems and their cytotoxicities, and to investigate the biochemical effect of the monomers on tyrosine phosphorylation, especially relating to the cell growth activity, of L929 cells in vitro. METHODS: The primers, uncured or cured adhesives (3M and Kuraray) were tested to determine the cytotoxicity of confluent L929 cells cultured by Eagle's MEM medium supplemented with 10% FCS. The area of cells affected by the eluted monomers were evaluated with an image analyzer and the concentrations of monomers eluted into the medium were measured with high performance liquid chromatography (HPLC) after 24h incubation. The protein composition of the stimulated cells was compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and tyrosine phosphorylation was detected by Western blot. RESULTS: The primer and uncured adhesives revealed variable cytotoxicities. 2-hydroxyethyl-methacrylate (HEMA) was the major component eluted from uncured primers and adhesives. Small amounts of triethylene glycol dimethacrylate (TEGDMA) were also detected from the uncured adhesives. The cytotoxicities of the adhesives decreased as photo activation time increased. The amount of monomers eluted from the cured adhesives was almost undetectable and did not reach a sufficient concentration to suppress cell viability or cell growth. The cytotoxicities of the primers and adhesives correlated well with the amounts of either HEMA or TEGDMA eluted. Moreover, a high concentration of HEMA (4 mg/ml medium) affected intracellular tyrosine phosphorylation, which is related to cellular activities. SIGNIFICANCE: Although the monomers present in dentin bonding resins are cytotoxic to L929 cells, the amount from cured bonding resin is very small and does not provide a cytotoxic dose. This data does however suggest that clinical exposure to the uncured primers and adhesives of dentin bonding resins should be minimized.
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Recubrimientos Dentinarios/toxicidad , Fibroblastos/efectos de los fármacos , Proteínas Tirosina Fosfatasas/efectos de los fármacos , Piel/efectos de los fármacos , Tirosina/efectos de los fármacos , Adhesivos/toxicidad , Análisis de Varianza , Animales , Western Blotting , Recuento de Células , División Celular/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Resinas Compuestas/toxicidad , Electroforesis en Gel de Poliacrilamida , Procesamiento de Imagen Asistido por Computador , Luz , Ensayo de Materiales , Metacrilatos/toxicidad , Ratones , Fosforilación , Polietilenglicoles/toxicidad , Ácidos Polimetacrílicos/toxicidad , Proteínas Tirosina Fosfatasas/análisis , Cementos de Resina/toxicidad , Piel/citología , Tirosina/análisisRESUMEN
The effect of dosing vehicle on toxicity and metabolism of unsaturated aliphatic nitriles was investigated in male Sprague-Dawley rats. Five unsaturated aliphatic nitriles--acrylonitrile, methacrylonitrile, allylnitrile, crotononitrile and fumaronitrile--were prepared in five different dosing vehicles--saline, corn oil, safflower oil, mineral oil, olive oil and Tween-20. Groups of six male rats were given 0.5 LD50 doses of the nitriles by gavage and they were observed for 12 It for cholinomimetic and central nervous system effects. Cyanide and glutathione levels were determined in blood and various organs at 1, 3 and 6 h after nitrile administration and thiocyanate levels were determined at 6 h after nitrile administration. The results indicate that all the vehicles studied potentiated the toxicity of all the nitriles compared to nitriles administered in saline and significantly increased their metabolism to cyanide and thiocyanate and nitrile-induced depletion of glutathione in rats. This behavior of vehicles illustrates the difficulty of identifying suitable vehicles for administration of lipophilic compounds in toxicology studies.