RESUMEN
Cholesterol esterase significantly contributes to cell membrane structure. It also facilitates transfer of cholesterol and phospholipids across membranes. Inhibition of this enzyme by a number of xenobiotics has been reported. This research sought to confirm if a widely used methacrylate monomer, bisphenol A dimethacrylate, inhibits porcine cholesterol esterase since this and other methacrylates are known to leach from various biomaterial preparations. A quantum mechanically developed computational chemistry model is presented. Specific chemical information linking potential mechanisms of cholesterol esterase inhibition to chemical structure is shown. Model chemical descriptors identified the importance of maximum oxygen valency and molecular shape/size to cholesterol esterase inhibition. A porcine cholesterol esterase inhibition mechanism is inherent in bisphenol A dimethacrylate which mimics chemical properties of reported cholesterol esterase inhibitors. This predictive semiempirical quantum mechanical model can be used to design new cholesterol esterase non-inhibitors for biocompatible biomaterials used in an aqueous environment.
Asunto(s)
Simulación por Computador , Esterol Esterasa/antagonistas & inhibidores , Porcinos , Animales , Modelos Moleculares , Conformación Molecular , Relación Estructura-Actividad Cuantitativa , Teoría Cuántica , Esterol Esterasa/metabolismoRESUMEN
Unpolymerized dental monomers can leach out into the oral biophase and are bioavailable for metabolism. We hypothesize that metabolites would be less toxic than parent monomers. We first identified the formation of metabolites from bisphenol F diglycidyl ether (BFDGE) and Bisphenol A glycidyl methacrylate (BISGMA) after their exposure to liver S9 fractions. Then, the metabolites and parent compounds were subjected to in vitro cytotoxicity, mutagenicity, and estrogenicity studies. Bisphenol A bis(2,3-dihydroxypropyl) ether and bisphenol F bis(2,3-dihydroxypropyl) ether were the hydroxylated metabolites of BISGMA and BFDGE, respectively. Cytotoxicity against L929 cells showed that the metabolites were significantly (p < 0.05) less cytotoxic than the parent monomers. Only BFDGE was mutagenic in the Ames assay with strain TA100 of Salmonella typhimurium. Parent and metabolite compounds did not stimulate estrogen-dependent MCF-7 cell proliferation above solvent controls. These results indicated that the hydroxylated metabolites were non-mutagenic, non-estrogenic, and less cytotoxic than their parent monomers.
Asunto(s)
Bisfenol A Glicidil Metacrilato/farmacocinética , Bisfenol A Glicidil Metacrilato/toxicidad , Materiales Dentales/metabolismo , Materiales Dentales/toxicidad , Compuestos Epoxi/farmacocinética , Compuestos Epoxi/toxicidad , Animales , Compuestos de Bencidrilo , Materiales Biocompatibles/metabolismo , Materiales Biocompatibles/toxicidad , Células Cultivadas/efectos de los fármacos , Estrógenos no Esteroides/farmacología , Humanos , Hidroxilación , Inactivación Metabólica , Células L/efectos de los fármacos , Ensayo de Materiales , Ratones , Microsomas Hepáticos/metabolismo , Pruebas de Mutagenicidad , Pruebas de ToxicidadRESUMEN
OBJECTIVES: Visible light cure oxirane/polyol resins of Cyracure UVR-6105 with pTHF-250 has been previously shown useful for development of dental composites. This oxirane/polyol (4016) in combination with other oxiranes were formulated into composites (4016E, 4016G and 4016GB) containing 72.9-74.9% quartz filler. The main objective of the study was to evaluate some of the physical properties and the biocompatibility of the composites. RESULTS: PhotoDSC analysis of composites demonstrated twice the enthalphy values of Z100 (31J/g). Composites 4016E and 4016G showed compressive strengths similar to Z100 (337+/-35Mpa), P>0.05. Discs of composite 4016E, containing Epon 825 oxirane (E), and composite 4016G containing Araldite GY 281 oxirane (G) were non-cytotoxic (-) while the composite 4016GB, containing G and Ebecryl 1830 (B), was mildly (+) cytotoxic to L929 cells in the agar diffusion assay. Seven-day extracts of 4016GB composite were cytotoxic while extracts of 4016E and 4016G were less cytotoxic to L929 cells in the MTT assay. Extracts were obtained from 7 day incubations of composite (3 cm(2) surface area/ml) in acetone or ethanol/saline (1:20) at 37 degrees C. All composite extracts were non-mutagenic to Ames strains TA100, TA98, TA97a and TA1535. The overall results with composite 4016GB suggest that leachable components were cytotoxic but non-mutagenic. With the exception of oxirane components, G and E, the oxirane Cyracure UVR-6105 and other components were non-mutagenic. From cytotoxicity studies, the photoinitiator, Sarcat CD 1012, was the most cytotoxic (TC(50)=14 microM) component. Components G (TC(50)=17 microM), E (TC(50)=50 microM) and B (TC(50)=151 microM) were significantly (p < 0.05) more cytotoxic than Cyracure UVR-6105 (1488 microM) and the polyol, pTHF-250 (TC(50)=6072 microM). SIGNIFICANCE: Favorable results obtained with composites 4016G and 4016E indicates that suitable oxirane/polyol formulations can be designed and optimized for development of dental composites with acceptable mechanical properties and biocompatibility. However, leachable analysis of extracts obtained from longer incubation periods is needed before final conclusions could be drawn about the leachability of oxirane components.
Asunto(s)
Materiales Biocompatibles/química , Materiales Biocompatibles/toxicidad , Resinas Compuestas/química , Resinas Compuestas/toxicidad , Óxido de Etileno/toxicidad , Animales , Área Bajo la Curva , Supervivencia Celular/efectos de los fármacos , Fenómenos Químicos , Química Física , Electroforesis en Gel de Agar , Resinas Epoxi/toxicidad , Células L/efectos de los fármacos , Dosificación Letal Mediana , Ensayo de Materiales , Ratones , Pruebas de Mutagenicidad , Polímeros/toxicidad , SolubilidadRESUMEN
The dental restorative monomer, BISGMA (2,2-bis[4-(2-hydroxy-3-methacryloxypropoxy)phenyl]propane), and bisphenol A diglycidyl ether (BADGE) increase the velocity of the reaction catalyzed by pancreatic cholesterol esterase (CEase, bovine). The metabolite of these monomers, bisphenol A bis(2,3-dihydroxypropyl) ether, and a common plasticizer, di-2-ethylhexyl phthalate (DEHP), also increase the velocity of CEase-catalyzed ester hydrolysis. BISGMA at concentrations of 1.5-8.0 microM increases the velocity to 126-169% of its value in the absence of BISGMA. Increasing BISGMA above 8 microM caused no further increase in velocity. BADGE at 7-25 microM increases the velocity to 112-205% of its value without BADGE. The metabolite of BISGMA and BADGE at concentrations of 2.0-7.1 microM increases the velocity to 103-113% of its value without metabolite. DEHP at concentrations of 0.52-4.3 microM increases the velocity to 108-187% of its value without DEHP. On the other hand, bisphenol A dimethacrylate is a competitive inhibitor of CEase, with a K(i) of 3.1 microM.
Asunto(s)
Recubrimientos Dentinarios/farmacología , Compuestos Epoxi/farmacología , Metacrilatos/farmacología , Esterol Esterasa/química , Compuestos de Bencidrilo , Butiratos/farmacología , Dietilhexil Ftalato/farmacología , Activación Enzimática/efectos de los fármacos , Cinética , Estructura Molecular , Esterol Esterasa/antagonistas & inhibidoresRESUMEN
This study addressed whether methacrylate monomers and polymers used in dentistry might degrade from enzymolysis by acetylcholinesterase (ACHE), cholesterol esterase (CHE), porcine liver esterase (PRLE), and a pancreatic lipase (PNL). Short (hour) and long-term (day) exposures were performed. Product ratios were used to determine surface hydrolysis of the polymeric materials. Enzyme kinetics were studied for the monomers when challenged by ACHE, CHE, and PRLE. In the case of PRLE, the V(max) for the dimethacrylate substrates varied slightly, but amounted to as much as 10% of that of p-nitrophenylacetate. The K(m) for triethylene glycol dimethacrylate (TEGDMA) was 197 microM for ACHE and 1107 microM for CHE. The V(max) was 2.7 nmol/min for ACHE and 3.5 nmol/min for CHE. TEGDMA was converted by CHE at 2% the rate of cholesteryl oleate. Long-term incubations of monomers with CHE and ACHE produced degrees of hydrolysis that evidenced structure dependency in the ability of the enzymes to effect hydrolysis. Particularly resistant were aromativ derivatives and those with branching in methacrylate linkages. Overall, the study confirms the ability of physiologically important esterases to catalyze the hydrolysis of biomaterial methacrylates.
Asunto(s)
Acetilcolinesterasa/metabolismo , Materiales Biocompatibles/metabolismo , Lipasa/metabolismo , Metacrilatos/metabolismo , Esterol Esterasa/metabolismo , Animales , Cromatografía Líquida de Alta Presión , Humanos , Hidrólisis , Cinética , Metacrilatos/química , Estructura Molecular , Polímeros/metabolismo , Factores de TiempoRESUMEN
Doxorubicin has a high affinity for inorganic iron, Fe(III), and has potential to form doxorubicin-Fe(III) complexes in biological systems. Indirect involvement of iron has been substantiated in the oxidative mutagenicity of doxorubicin. In this study, however, direct involvement of Fe(III) was evaluated in mutagenicity studies with the doxorubicin-Fe(III) complex. The Salmonella mutagenicity assay with strain TA102 was used with a pre-incubation step. The highest mutagenicity of doxorubicin-Fe(III) complex was observed at the dose of 2.5nmol/plate of the complex. The S9-mix decreased this highest mutagenicity but increased the number of revertants at a higher dose of 10nmol/plate of the complex. On the other hand, the mutagenicity of the doxorubicin-Fe(III) complex at the doses of 0.25, 0.5, 1 and 2nmol/plate was enhanced about twice by the addition of glutathione plus H(2)O(2). This enhanced mutagenicity as well as of the complex itself, the complex plus glutathione, and the complex plus H(2)O(2) were reduced by the addition of ADR-529, an Fe(III) chelator, and potassium iodide, a hydroxyl radical scavenger. These results indicate that doxorubicin-Fe(III) complex exert the mutagenicity through oxidative DNA damage and that Fe(III) is a required element in the mutagenesis of doxorubicin.
Asunto(s)
Daño del ADN , ADN Bacteriano/efectos de los fármacos , Doxorrubicina/toxicidad , Compuestos Organometálicos/toxicidad , Salmonella typhimurium/efectos de los fármacos , Animales , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Depuradores de Radicales Libres/farmacología , Radicales Libres , Genes Bacterianos/efectos de los fármacos , Glutatión/toxicidad , Peróxido de Hidrógeno/toxicidad , Quelantes del Hierro/farmacología , Masculino , Microsomas Hepáticos/metabolismo , Estructura Molecular , Pruebas de Mutagenicidad , Oxidación-Reducción , Yoduro de Potasio/farmacología , Ratas , Ratas Sprague-Dawley , Razoxano/farmacología , Salmonella typhimurium/genéticaRESUMEN
Many reports have demonstrated inflammation after the placement of dental restorations. To explain this side-effect, we studied a biomarker in the inflammatory response. The intercellular adhesion molecule-1 (ICAM-1) is a key mediator for recruitment of leukocytes to the site of inflammation. Therefore, we investigated whether methacrylates (a BISGMA-based dental resin, BISGMA, and MAA) and Cyracure UVR 6105, an epoxy monomer, could alter ICAM-1 expression in unstimulated and TNF-alpha-stimulated endothelial cells. Six-well plates with monolayers of human umbilical vein cells, ECV 304 (ATCC CRL 1998), were exposed to TNF-alpha (1 ng/mL) in the presence and absence of subtoxic and TC50 doses of chemicals for 24 hrs at 37 degrees C/5% CO2. Several doses of TNF-alpha (0.5-2 ng/mL) were coincubated with 100 microL of undiluted aqueous dental resin extracts. Cells were harvested and stained with mAB FITC-conjugated anti-human ICAM-1 (CD54). ICAM-1 expression was measured by flow cytometry. Cells expressed basal levels of ICAM-1, which was up-regulated by TNF-alpha but was not changed by all samples studied. Except for UVR 6105, the methacrylates significantly decreased ICAM-1 expression in TNF-alpha-stimulated cells. These findings suggest that methacrylates may decrease the recruitment of leukocytes to sites of inflammation.
Asunto(s)
Materiales Dentales/toxicidad , Endotelio/efectos de los fármacos , Molécula 1 de Adhesión Intercelular/biosíntesis , Resinas Sintéticas/toxicidad , Factor de Necrosis Tumoral alfa/farmacología , Análisis de Varianza , Bisfenol A Glicidil Metacrilato/toxicidad , Línea Celular Transformada , Supervivencia Celular/efectos de los fármacos , Resinas Compuestas/toxicidad , Ácidos Ciclohexanocarboxílicos/toxicidad , Relación Dosis-Respuesta a Droga , Endotelio/citología , Endotelio/metabolismo , Humanos , Técnicas In Vitro , Leucocitos/efectos de los fármacos , Metacrilatos/toxicidad , Dióxido de Silicio/toxicidad , Circonio/toxicidadRESUMEN
In development of photopolymerized expanding monomers with epoxy resin systems, there is a need for reactive expanding monomers that exert a good biocompatibility profile. The objective of this study was to evaluate the in vitro toxicology of new spiroorthocarbonates designed to be expanding monomers. The expanding monomers investigated were: trans/trans-2,3,8,9-di(tetramethylene)-1,5,7,11-tetraoxaspiro[5,5] undecane (DTM-TOSU), 5,5-diethyl-19-oxadispiro-[1,3-dioxane-2,2'-1,3-dioxane-5',4'-bicy clo[4.1.0]heptane] (DECHE-TOSU); 3,9-diethyl-3,9-dipropionyloxy methyl-1,5,7,11-tetraoxaspiro[5.5]undecane (DEDPM-TOSU); and 3,9-diethyl-3,9-diacetoxy methyl-1,5,7,11-tetraoxaspiro[5.5]undecane (DAMDE-TOSU). The in vitro toxicology of these monomers measured their cytotoxicity and mutagenicity potential. Succinic dehydrogenase (SDH) activity in the MTT assay was used to assess the toxic dose that kills 50% of cells (TC50) for all the monomers. Their mutagenic potential was measured in the Ames Salmonella assay with and without metabolic activation. Two solvents, DMSO and acetone, were used to validate effects. Appropriate controls included the solvents alone. All the expanding monomers in this study were less cytotoxic than BISGMA (p < 0.01), a commercial component of dental restoratives. The relative cytotoxicity of the expanding monomers in DMSO was defined in the following order: DTM-TOSU (more toxic) > DECHE-TOSU > DEDPM-TOSU > DAMDE-TOSU. Each was significantly different from the other (p < 0.05). Overall, the TC50 values of all expanding monomers were significantly greater in DMSO than in acetone (p < 0.05). However, for BISGMA this trend was opposite. For mutagenicity results, the expanding monomers were non-mutagenic and there was no solvent effect on this outcome. The non-mutagenicity and low cytotoxicity profile of these expanding monomers suggests their potential for development of biocompatible non-shrinking composites.
Asunto(s)
Carbonatos/química , Resinas Compuestas/toxicidad , Compuestos de Espiro/toxicidad , Animales , Línea Celular/efectos de los fármacos , Ratones , Pruebas de Mutagenicidad/métodos , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genética , Sales de Tetrazolio/análisis , Sales de Tetrazolio/farmacología , Pruebas de Toxicidad/métodosRESUMEN
Bisphenol A (BPA) is a common structural component in a wide variety of biomaterial monomers. The effects of BPA and the following derivatives: bisphenol A glycidyl methacrylate (BisGMA), bisphenol A glycidyl diacrylate (BAGDA), bisphenol A ethoxylate dimethacrylate (BAEDM), bisphenol A dimethacrylate (BADM), and bisphenol A diglycidyl ether (BADGE) on mixed function oxidases (MFOs) are reported in this study. The rate of formation of metabolites from isoform-specific substrates for the MFOs (or cytochromes) CYP 1A, 2A, 2C, 2E, 3A, and 4A in the absence (control) and presence of BPA and derivatives was used to assess inhibition or stimulation of human, rat (male and female) liver, and minipig liver microsomal MFO activity. For human preparations the strongest inhibition by BPA was observed for CYP 2C. The inhibition was most prominent when a lower dose of BPA was used on the complete post-mitochondrial fraction. BPA inhibited rat microsomal CYP 1A isoform-specific metabolite production to 29 +/- 3% of control levels (100%). Biomaterial monomers exhibited mixed effects. For example, BPA stimulated CYP 4A in pooled human S9 to 129 +/- 1% of control. Also, BADM and BAGDA stimulated CYP 4A to 141% and 142% of control values, respectively.
Asunto(s)
Inhibidores Enzimáticos del Citocromo P-450 , Inhibidores Enzimáticos/farmacología , Fenoles/farmacología , Animales , Compuestos de Bencidrilo , Cafeína/química , Cumarinas/química , Sistema Enzimático del Citocromo P-450/química , Remoción de Radical Alquila , Diclofenaco/química , Inhibidores Enzimáticos/química , Humanos , Isoenzimas/antagonistas & inhibidores , Isoenzimas/química , Hígado/metabolismo , Masculino , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/enzimología , Fenoles/química , Ratas , Ratas Sprague-Dawley , Especificidad de la Especie , Porcinos , Porcinos EnanosRESUMEN
OBJECTIVE: The objective of this study was to evaluate the effect of adding a spiroorthocarbonate (SOC) or a polyol on the cytotoxicity of epoxy-based dental resins. METHODS: Resins contained one of the epoxies: diglycidyl ether Bisphenol A (GY-6004); 3,4-epoxycyclohexanemethyl-3,4-epoxycyclohexane carboxylate (UVR-6105); vinyl cyclohexane dioxide (ERL-4206) or the three-epoxy mixture (Epoxy-M). The SOC was t/t-2,3,8,9-di(tetramethylene)-1,5,7,11-tetraoxaspiro[5.5]undecane (SOC). The polyols were polytetrahydrofuran (p-THF-250) and polycaprolactone triol (TONE-301). The photoinitiator (4-octylphenyl)phenyliodonium hexafluoroantimonate and camphorquinone were used for light curing the resins. Four types of resins (epoxy, SOC/epoxy, polyol/epoxy and SOC/polyol/epoxy) were evaluated for cytotoxicity as solids in the agar diffusion assay and as aqueous extracts in the MTT assay using L929 cells. RESULTS: In agar diffusion analysis, ERL-4206 and UVR-6105 resins were severely cytotoxic (+3), but the addition of SOC changed them to non-cytotoxic (-). Addition of 1-3% SOC changed Epoxy-M from mild (+) to non-cytotoxic. Adding SOC changed GY-6004 from moderate (+2) to mild (-) cytotoxicity. Generally, addition of SOC did not change cytotoxicity when added to polyol/epoxy combinations. Either polyol produced resins with reduced cytotoxicity when added to UVR-6105, but the opposite occurred when added to Epoxy-M resins. In MTT analysis, percent cell survival from 100 microliters resin extracts were statistically compared (ANOVA, p < 0.05). Epoxy-M and GY-6004 resin extracts were significantly less cytotoxic than UVR-6105 and ERL-4206 resin extracts were. Overall, the SOC component reduced the cytotoxicity of all SOC/epoxy combinations, except SOC/ERL-4206, which was significantly more cytotoxic than ERL-4206 resin extract. This may be the result of cell fixative effects observed for SOC/ERL-4206 in agar diffusion analysis. Addition of SOC produced significantly less cytotoxic SOC/polyol/Epoxy-M resins when compared to its non-SOC counterpart. The contrary result was obtained with SOC/polyol/UVR-6105 resin combinations. Consistent with agar diffusion results, adding polyol significantly decreased cytotoxicity of UVR-6105 resins. The cytotoxicity of these resins may be related to the 50% cytotoxicity (TC50) of their components as leachates. The TC50 values of the individual components were compared to BISGMA. Polyols, epoxy monomers, SOC monomer and camphorquinone were significantly (p < 0.05) less cytotoxic than BISGMA. SIGNIFICANCE: Addition of SOCs and polyols in the formulation of epoxy-based resins may contribute to development of biocompatible dental composites.
Asunto(s)
Materiales Dentales/toxicidad , Resinas Epoxi/toxicidad , Análisis de Varianza , Animales , Bioensayo , Materiales Dentales/química , Resinas Epoxi/química , Concentración 50 Inhibidora , Células L/efectos de los fármacos , Ensayo de Materiales , Ratones , Polímeros/toxicidad , Compuestos de Espiro/toxicidadRESUMEN
OBJECTIVES: The purpose of this study was to compare a methylthiazole tetrazolium (MTT) dye colorimetric method with the standard 51Cr assay as methods of assessing cytotoxicity of dental materials. METHODS: Two MTT-based colorimetric formats, test tube and 96-well microplate methods, were compared to the 51Cr release assay. A series of eight dental materials were evaluated. Cytotoxicity profiles were determined for each test material. A TC50 value (Toxic Concentration required to kill 50% of the cells) was determined for each biomaterial, and these results were used to make statistical comparisons between the methods. RESULTS: The three methods were statistically correlated (p<0.005) by comparison of the eight samples tested. That is, the same rank in toxicity was given by the two tetrazolium sample formats and the 51Cr method. SIGNIFICANCE: The MTT assay was found to have several advantages in comparison to the current standard 51Cr release assay. Optimized in the 96-well format, complete dose response curves and greater sample comparisons can be made rapidly, making the MTT method more economical in time and cost. Furthermore, the MTT method is based on intracellular biochemical changes, measuring cell viability rather than cell morbidity, and has lower detectable limits than the 51Cr release method. There is also less detector chemical binding interference than encountered in the 51Cr release method.
Asunto(s)
Materiales Dentales/toxicidad , Evaluación Preclínica de Medicamentos/métodos , Pruebas de Toxicidad/métodos , Animales , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Cromatos/metabolismo , Radioisótopos de Cromo/metabolismo , Colorimetría , Relación Dosis-Respuesta a Droga , Indicadores y Reactivos , Células L , Ratones , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Compuestos de Sodio/metabolismo , Sales de Tetrazolio , Tiazoles , Pruebas de Toxicidad/instrumentaciónRESUMEN
A major improvement in dental restoratives is possible through the development of biomaterials that do not shrink upon polymerization, hence, avoid leakage and subsequent breakdown. Polymers containing spiroorthocarbonates (SOCs) show promise in this respect, but their toxicology in copolymerized materials has not been explored. In this study, the in vitro toxicology of these materials in homopolymer form and in two trial non-shrinking epoxy co-polymers was evaluated for cytotoxicity and mutagenicity. Cytotoxicity was determined by the MTT test to measure the lethality effect on mouse L929 cells. Mutagenicity was evaluated using the Ames-Salmonella Test. For comparison, commercial composite and adhesive materials as well as several other materials of current interest in dentistry were also evaluated. Epoxy resin samples containing 5% of either T/T SOC or Dp SOC reduced the cytotoxicity (TC50) from approximately 400 to 800 micrograms/200 microliters. The epoxy-spiro copolymers had more favorable TC50 values than the commercial product Super-Bond. They showed TC50 values on the order of 35% greater than Super-Bond and 45% less than Scotchbond 2, the latter two being materials currently used in the clinic. These two comparatives demonstrated dose response curves with lower doses at maximum cell kill values than the spiro materials. The epoxy formulations all showed weak mutagenesis, but this is attributed to the epoxy formulation and not the SOCs. Although considerable toxicology is yet be conducted, these in vitro results suggest that biocompatible copolymer formulations for spiroorthocarbonates are a developmental reality.
Asunto(s)
Recubrimiento Dental Adhesivo , Recubrimientos Dentinarios/toxicidad , Resinas Epoxi/toxicidad , Compuestos de Espiro/toxicidad , Adhesivos , Animales , Materiales Biocompatibles , Carbonatos/toxicidad , Línea Celular , Supervivencia Celular , Células Cultivadas , Recubrimientos Dentinarios/química , Fibroblastos/efectos de los fármacos , Ratones , Pruebas de Mutagenicidad , Compuestos de Espiro/químicaRESUMEN
Current in vitro biocompatibility methods do not evaluate the degradation of biomaterials after contact with enzymes that might be present in the oral or systemic environment. In this study, two methods of in vitro enzyme degradation and a method for the separation of the degradative products by high performance thin-layer chromatography (HPTLC) are reported. In the first method two dental adhesives, Scotchbond and Scotchbond II, and two dental composites, Heliomolar and P-50, were evaluated. These materials were incubated with four different enzymatic preparations for periods of up to 72 h. The enzymes were lipase, esterase, and liver enzyme extracts from both mouse and rat. Chloroform soluble products extracted from the aqueous phase were examined by HPTLC for decomposition products resulting from enzyme activity. The second method was similar, but analyzed the aqueous fraction directly without chloroform extraction. In this method five dental restorative materials, P-50, P-30, Scotchbond II, Silux, and Silux Plus, were incubated with a nonspecific porcine liver esterase. In addition to the polymerized biomaterials, monomers containing methacrylic acid units were also hydrolyzed with esterase and analyzed by ion chromatography to establish the sensitivity of the enzyme simulator. Each biomaterial presented thin-layer zones not present before enzymatic action. These experiments provide support that aqueous enzymatic action may facilitate the hydrolytic weakening of polymeric biomaterials.
Asunto(s)
Materiales Biocompatibles/metabolismo , Esterasas/metabolismo , Ensayo de Materiales/métodos , Metacrilatos/química , Animales , Cromatografía Líquida de Alta Presión/métodos , Cromatografía en Capa Delgada/métodos , Materiales Dentales/metabolismo , Semivida , Hidrólisis , Lipasa/metabolismo , Ratones , Modelos Biológicos , Polímeros/metabolismo , RatasRESUMEN
There is a presumption that copper and anthracycline drugs will interact with DNA to produce genotoxicity. This is of concern because serum copper levels are increased in certain neoplastic diseases. To test the interaction, it was determined if the metal ion could alter the mutagenesis of doxorubicin and related drugs in the Salmonella microsome test. In the standard form of the test, doxorubicin was strongly mutagenic against frameshift-sensitive strain TA98. When cupric acetate was added with doxorubicin it amplified the mutagenesis of the antineoplastic with an increase of approximately 19% in peak mutagenic values. This apparent "chemoactivation" was evaluated by additional applications of the Salmonella test. Preincubation of cupric acetate, drug, and bacteria (+/- rat liver S9 fraction) also resulted in a copper-amplifying effect. In the preincubation method copper produced a drug concentration-related increase of more than 700% in the mutagenicity of doxorubicin. This large an increase occurred without S9 in the test. The effects observed in TA98 were not seen with TA102, a strain sensitive to oxidation mechanisms. Copper amplification in the mutagenicity of a positive control, aflatoxin B1, was also observed with TA98 but these effects were not seen with the chelator, EDTA, the antifolate antineoplastic drug, methotrexate, or a test negative amino acid, methionine. Results point to a direct frameshift mechanism to explain the increase in mutagenicity with copper. Amplification of mutagenicity found in this study provides initial experimental support that anthracycline-metal ion-DNA associations might contribute to genotoxicity as has been inferred in the literature.
Asunto(s)
Cobre/farmacología , Doxorrubicina/toxicidad , Mutágenos/toxicidad , Cobre/química , Relación Dosis-Respuesta a Droga , Doxorrubicina/análogos & derivados , Doxorrubicina/química , Sinergismo Farmacológico , Concentración de Iones de Hidrógeno , Pruebas de Mutagenicidad , Salmonella/efectos de los fármacos , Espectrofotometría UltravioletaRESUMEN
The mutagenicities of [1R] and [1S] aflatoxicol were measured using the Salmonella microsome test. In strain TA100 the [1R] form (unnatural aflatoxicol, aflatoxicol B) had a mutagenic potency approximately four times that of the [1S] epimer (natural aflatoxicol, aflatoxicol A, Ro) in the presence of S-9 liver microsomal fraction. The order in mutagenic potency compared to some other toxicologically important aflatoxins was as follows: B1 greater than [1R] approximately equal to G1 much greater than [1S] much much greater than B2. Thus, the trans relationship between the vinyl ether and hydroxyl groups leads to greater mutagenicity than the cis relationship. This may be important in the elucidation of stereochemical structure-activity relationships for the aflatoxins.
Asunto(s)
Aflatoxinas/farmacología , Mutágenos , Mutación , Salmonella typhimurium/efectos de los fármacos , Animales , Biotransformación , Microsomas Hepáticos/metabolismo , Pruebas de Mutagenicidad , Ratas , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
The metabolism of aflatoxin B1 by the post-mitochondrial fraction of human autopsy or biopsy liver and monkey necropsy liver was investigated. Aflatoxin Q1, a hydroxy metabolite of B1, was produced by 18 of 22 human liver samples and by 5 of 7 species of non-human primates investigated. Human specimens from both sexes, ages 20-80 years, with a variety of hepatic histological and historical influences produced this metabolite. Aflatoxin Q1 was of variable quantitative importance. Yields seldom exceeded 10% of initial B1. However, based upon its persistent formation in vitro, these studies identify the need to clarify the role of Q1 in aflatoxin epidemiology.
Asunto(s)
Aflatoxinas/metabolismo , Hígado/metabolismo , Adulto , Aflatoxina B1 , Anciano , Anciano de 80 o más Años , Animales , Femenino , Humanos , Masculino , Persona de Mediana Edad , Primates/metabolismo , Especificidad de la Especie , Fracciones Subcelulares/metabolismoRESUMEN
The non-microsomal mutagenesis of a series of aflatoxins in Salmonella typhimurium TA100 and TA98 was evaluated. Aflatoxin B1 had the highest direct or non-microsomal potency. However, the order of direct mutagenic activity as compared to indirect or microsomal activity was as follows; aflatoxin Q1 much greater than G1 greater than B1 greater than aflatoxicol. This order was determined using a new plotting strategy for identification of direct contribution to the mutagenesis, an I-D graph (indirect vs. direct activities). It is suggested from our analyses that the fused carbonyl bearing rings in aflatoxins are necessary for direct mutagenesis. This activity is facilitated by oxygen substitution at the terminal carbonyl ring. On the other hand, the cyclopentene structure may either attenuate or amplify the microsome mediated mutagenesis which is assumed to be produced by the vinyl ether portion of the molecule. This was dependent upon substitution at the cyclopentene. Stereo-sensitivity was revealed by the greater mutagenicity of unnatural aflatoxicol over the natural epimer. These observations point to a need for further study of direct binding and transmolecular-structure-activity relationships in order to fully develop the mechanism of aflatoxin mutagenicity.
Asunto(s)
Aflatoxinas/toxicidad , Mutágenos , Salmonella typhimurium/genética , Aflatoxina B1 , Aflatoxinas/metabolismo , Animales , Técnicas In Vitro , Indicadores y Reactivos , Pruebas de Mutagenicidad , Mutágenos/metabolismo , Ratas , Relación Estructura-ActividadRESUMEN
An investigation of the mutagenicity of aflatoxin Q1 in the Salmonella-mutagenicity test is reported. This mammalian metabolite of aflatoxin B1 was marginally mutagenic to strain TA 98 when either rat or human liver microsomes were present in the test. However, it was mutagenic to TA 100 with the same liver activation sources in the assay. Moreover, it was mutagenic to TA 100 in the absence of liver microsomes. This Q1 mutagenesis, presumably a direct acting base substitution effect, was nearly equal to the liver activated mutagenesis. It was also greater in this respect than aflatoxin B1 or aflatoxicol.
Asunto(s)
Aflatoxinas/toxicidad , Mutágenos , Aflatoxina B1 , Aflatoxinas/metabolismo , Animales , Biotransformación , Humanos , Técnicas In Vitro , Microsomas Hepáticos/metabolismo , Pruebas de Mutagenicidad , Ratas , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genéticaRESUMEN
The investigation of the in vitro metabolism of aflatoxin Q1 by the post-mitochondrial fraction of mouse and rabbit liver is reported. Both animals metabolized this substance at a turnover similar to aflatoxin B1. There was a higher bound fraction and lower aqueous fraction from the metabolism of B1 than from aflatoxin Q1. The aqueous fraction of the metabolisms evidenced the beta-D-glucuronide of aflatoxin Q1. The rabbit metabolism of Q1 was characterized by high levels of chloroform soluble metabolites. In contrast, the mouse metabolism resulted in high glucuronide and bound Q1 metabolite levels.
Asunto(s)
Aflatoxinas/metabolismo , Hígado/metabolismo , Animales , Arocloros/farmacología , Biotransformación , Cromatografía Líquida de Alta Presión , Cromatografía en Capa Delgada , Inducción Enzimática/efectos de los fármacos , Hidrólisis , Técnicas In Vitro , Masculino , Ratones , Proteínas/metabolismo , Conejos , Fracciones Subcelulares/enzimologíaRESUMEN
An investigation of the in vitro biotransformation of aflatoxin Q1 by the post-mitochondrial fraction of rat liver is reported. These studies showed a 10 to 25 percent turnover of aflatoxin Q1, with the subsequent formation of five metabolites. A major metabolite was determined to be the beta-D-glucuronide of aflatoxin Q1.