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1.
Ecotoxicol Environ Saf ; 281: 116606, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38896907

RESUMEN

Finasteride, a steroid 5-alpha reductase inhibitor, is commonly used for the treatment of benign prostatic hyperplasia and hair loss. However, despite continued use, its environmental implications have not been thoroughly investigated. Thus, we investigated the acute and chronic adverse impacts of finasteride on Daphnia magna, a crucial planktonic crustacean in freshwater ecosystems selected as bioindicator organism for understanding the ecotoxicological effects. Chronic exposure (for 23 days) to finasteride negatively affected development and reproduction, leading to reduced fecundity, delayed first brood, reduced growth, and reduced neonate size. Additionally, acute exposure (< 24 h) caused decreased expression levels of genes crucial for reproduction and development, especially EcR-A/B (ecdysone receptors), Jhe (juvenile hormone esterase), and Vtg2 (vitellogenin), with oxidative stress-related genes. Untargeted lipidomics/metabolomic analyses revealed lipidomic alteration, including 19 upregulated and 4 downregulated enriched lipid ontology categories, and confirmed downregulation of metabolites. Pathway analysis implicated significant effects on metabolic pathways, including the pentose phosphate pathway, histidine metabolism, beta-alanine metabolism, as well as alanine, aspartate, and glutamate metabolism. This comprehensive study unravels the intricate molecular and metabolic responses of D. magna to finasteride exposure, underscoring the multifaceted impacts of this anti-androgenic compound on a keystone species of freshwater ecosystems. The findings emphasize the importance of understanding the environmental repercussions of widely used pharmaceuticals to protect biodiversity in aquatic ecosystems.


Asunto(s)
Inhibidores de 5-alfa-Reductasa , Daphnia , Finasterida , Metabolismo de los Lípidos , Contaminantes Químicos del Agua , Animales , Finasterida/toxicidad , Daphnia/efectos de los fármacos , Inhibidores de 5-alfa-Reductasa/toxicidad , Contaminantes Químicos del Agua/toxicidad , Metabolismo de los Lípidos/efectos de los fármacos , Disruptores Endocrinos/toxicidad , Reproducción/efectos de los fármacos , Lipidómica , Daphnia magna
2.
Curr Issues Mol Biol ; 46(6): 5551-5560, 2024 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-38921003

RESUMEN

Extensive research on medicinal herbs for bioactive compounds proposes that they could replace synthetic drugs, reducing side effects and economic burdens. Especially, interest in the synergistic benefits of natural products is increasing, implying that their combined use may enhance therapeutic effectiveness. This study aimed to explore the synergetic effects of turmeric (Curcuma longa L.) and black pepper (Piper nigrum L.) extract on lung normal (MRC-5) and cancer (A549 and NCI-H292) cell lines. The turmeric extract (TM) only affected the lung cancer cell lines, but it had no impact on the MRC-5 cell line. On the other hand, the black pepper extract (BP) did not cause any damage to either the lung normal or cancer cell lines, even at concentrations of up to 400 µg/mL. Response surface methodology was used to predict the ideal synergistic concentrations (EC50) of TM and BP, which were found to be 48.5 and 241.7 µg/mL, respectively. Notably, the selected condition resulted in higher cytotoxicity compared to the exposure to TM alone, indicating a potent synergetic effect. The rate of curcumin degradation under this combined treatment was significantly decreased to 49.72 ± 5.00 nmol/h/µg for A549 cells and 47.53 ± 4.78 nmol/h/µg for NCI-H292 cells, respectively, as compared to curcumin alone. Taken together, this study confirmed the potent synergistic effect of TM and BP on lung cancer cell lines. Further research is required to identify their specific synergetic mechanisms. Our findings provide crucial foundational data on the synergistic effects of TM and BP.

3.
Toxicol In Vitro ; 98: 105838, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38710238

RESUMEN

Interactions between endocrine-disruptor chemicals (EDCs) and androgen receptor (AR) have adverse effects on the endocrine system, leading to human reproductive dysfunction. Bisphenol A (BPA) is an EDC that can damage both the environment and human health. Although numerous BPA analogues have been produced as substitutes for BPA, few studies have evaluated their endocrine-disrupting abilities. We assessed the (anti)-androgenic activities of BPA and its analogues using a yeast-based reporter assay. The BPA analogues tested were bisphenol S (BPS), 4-phenylphenol (4PP), 4,4'-(9-fluorenyliden)-diphenol (BPFL), tetramethyl bisphenol F (TMBPF), and tetramethyl bisphenol A (TMBPA). We also conducted molecular docking and dynamics simulations to assess the interactions of BPA and its analogues with the ligand-binding domain of human AR (AR-LBD). Neither BPA nor its analogues had androgenic activity; however, all except BPFL exerted robust anti-androgenic effects. Consistent with the in vitro results, anti-androgenic analogues of BPA formed hydrogen bonding patterns with key residues that differed from the patterns of endogenous hormones, indicating that the analogues display in inappropriate orientations when interacting with the binding pocket of AR-LBD. Our findings indicate that BPA and its analogues disrupt androgen signaling by interacting with the AR-LBD. Overall, BPA and its analogues display endocrine-disrupting activity, which is mediated by AR.


Asunto(s)
Compuestos de Bencidrilo , Disruptores Endocrinos , Simulación del Acoplamiento Molecular , Fenoles , Receptores Androgénicos , Fenoles/toxicidad , Fenoles/química , Compuestos de Bencidrilo/toxicidad , Compuestos de Bencidrilo/química , Receptores Androgénicos/metabolismo , Receptores Androgénicos/efectos de los fármacos , Disruptores Endocrinos/toxicidad , Disruptores Endocrinos/química , Humanos , Simulación por Computador , Sulfonas/toxicidad , Sulfonas/química , Andrógenos/química
4.
Antioxidants (Basel) ; 13(2)2024 Feb 03.
Artículo en Inglés | MEDLINE | ID: mdl-38397793

RESUMEN

Photoaging refers to the accumulation of skin damage which includes wrinkle formation, loss of elasticity, and epidermal thickening due to repeated ultraviolet (UV) irradiation. The present study investigated the protective effects of Elaeagnus umbellata fruit extract (Elaea) on UV-mediated photoaged skin of SKH1 hairless mice and compared the effects of Elaea with ascorbic acid. Although there was no difference in body weight between groups during experimental period, oral administration of 50-200 mg/kg Elaea once daily for 15 weeks significantly prevented an increase in skin weight, epithelial thickening of epidermis, and apoptosis caused by UV irradiation. Skin replica and histopathological analyses revealed that Elaea dose-dependently decreased wrinkle and microfold formation. In addition, Elaea administration restored UV-mediated reduction in type I collagen and hyaluronan through the inhibition of matrix metalloproteinases and p38 mitogen-activated protein kinase expression. Moreover, Elaea suppressed UV-dependent increases in superoxide anion production, fatty acid oxidation, and protein nitration by up-regulating antioxidant system. Furthermore, Elaea alleviated infiltration of inflammatory cells in UV-irradiated skin. The preventive effects of 100 mg/kg Elaea administration against UV-induced photoaging were similar to those by 100 mg/kg ascorbic acid. Collectively, the present study suggests that the E. umbellata fruit is a promising edible candidate to prevent skin photoaging.

5.
Front Pharmacol ; 14: 1067408, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36874001

RESUMEN

The SARS-CoV-2 pandemic requires a new therapeutic target for viral infection, and papain-like protease (Plpro) has been suggested as a druggable target. This in-vitro study was conducted to examine the drug metabolism of the GRL0617 and HY-17542, Plpro inhibitors. Metabolism of these inhibitors was studied to predict the pharmacokinetics in human liver microsomes. The hepatic cytochrome P450 (CYP) isoforms responsible for their metabolism were identified using recombinant enzymes. The drug-drug interaction potential mediated by cytochrome P450 inhibition was estimated. In human liver microsomes, the Plpro inhibitors had phase I and phase I + II metabolism with half-lives of 26.35 and 29.53 min, respectively. Hydroxylation (M1) and desaturation (-H2, M3) of the para-amino toluene side chain were the predominant reactions mediated with CYP3A4 and CYP3A5. CYP2D6 is responsible for the hydroxylation of the naphthalene side ring. GRL0617 inhibits major drug-metabolizing enzymes, including CYP2C9 and CYP3A4. HY-17542 is structural analog of GRL0617 and it is metabolized to GRL0617 through non-cytochrome P450 reactions in human liver microsomes without NADPH. Like GRL0617 and HY-17542 undergoes additional hepatic metabolism. The in-vitro hepatic metabolism of the Plpro inhibitors featured short half-lives; preclinical metabolism studies are needed to determine therapeutic doses for these inhibitors.

6.
Small ; 19(25): e2300236, 2023 06.
Artículo en Inglés | MEDLINE | ID: mdl-36932895

RESUMEN

The disruption of thyroid hormones because of chemical exposure is a significant societal problem. Chemical evaluations of environmental and human health risks are conventionally based on animal experiments. However, owing to recent breakthroughs in biotechnology, the potential toxicity of chemicals can now be evaluated using 3D cell cultures. In this study, the interactive effects of thyroid-friendly soft (TS) microspheres on thyroid cell aggregates are elucidated and their potential as a reliable toxicity assessment tool is evaluated. Using state-of-the-art characterization methods coupled with cell-based analysis and quadrupole time-of-flight mass spectrometry, it is shown that TS-microsphere-integrated thyroid cell aggregates exhibit improved thyroid function. Specifically, the responses of zebrafish embryos, which are used for thyroid toxicity analysis, and the TS-microsphere-integrated cell aggregates to methimazole (MMI), a known thyroid inhibitor, are compared. The results show that the thyroid hormone disruption response of the TS-microsphere-integrated thyroid cell aggregates to MMI is more sensitive compared with those of the zebrafish embryos and conventionally formed cell aggregates. This proof-of-concept approach can be used to control cellular function in the desired direction and hence evaluate thyroid function. Thus, the proposed TS-microsphere-integrated cell aggregates may yield new fundamental insights for advancing in vitro cell-based research.


Asunto(s)
Glándula Tiroides , Pez Cebra , Animales , Humanos , Antitiroideos/farmacología , Hormonas Tiroideas/farmacología , Metimazol/toxicidad
7.
Ecotoxicol Environ Saf ; 243: 113965, 2022 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-35994907

RESUMEN

Several phenol derivatives are suspected endocrine disruptors and have received attention in risk assessment studies for several decades owing to the structural similarity between estrogens and phenolic compounds. We assessed the endocrine disrupting effect of the phenolic compound para-phenylphenol (PPP) through acute tests and evaluating chronic endpoints in an invertebrate model, Daphnia magna. Exposure of D. magna to PPP induced substantial adverse effects, namely, reduced fecundity, slowed growth rate, delayed first brood, and a reduction in neonate size. Furthermore, we investigated the mRNA expression of relevant genes to elucidate the mechanism of endocrine disruption by PPP. Exposure of D. magna to PPP induced the substantial downregulation of genes and markers related to reproduction and development, such as EcR-A, EcR-B, Jhe, and Vtg. Consequently, we demonstrated that PPP has an endocrine disrupting effect on reproduction and development in D. magna.


Asunto(s)
Disruptores Endocrinos , Contaminantes Químicos del Agua , Animales , Daphnia , Disruptores Endocrinos/toxicidad , Sistema Endocrino , Reproducción , Contaminantes Químicos del Agua/toxicidad
8.
Environ Sci Pollut Res Int ; 29(37): 55639-55650, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-35320476

RESUMEN

Mono(2-ethylhexyl) phthalate (MEHP) is a primary metabolite of di(2-ethylhexyl) phthalate (DEHP), which is widely used in industry as a plasticizer. Both DEHP and MEHP have been identified as endocrine disruptors affecting reproduction systems in natural aquatic environments. However, the effects of MEHP exposure on aquatic invertebrates such as Daphnia magna are still poorly understood. In the present study, lipid alterations caused by MEHP in D. magna were identified by analyzing lipid accumulation and nontarget metabolomics. In addition, reproductive endpoints were investigated. MEHP exposure under any conditions upto 2 mg/L was not associated with mortality of D. magna; yet, the number of lipid droplets and the adult female daphnids reproduction rates increased after 96 h of exposure and 21 days of exposure, respectively. MEHP also enhanced lipid metabolism, as evident from 283 potential lipid metabolites, including glycerolipids, glycerophospholipids, and sphingolipids, identified following 48 h of exposure. The MEHP-treated group exhibited significantly higher ecdysone receptor (EcR) and vitellogenin 2 (Vtg2) expression levels at 6 and 24 h. At 48 h, EcR and Vtg2 expression levels were downregulated in the 1 and 2 mg/L MEHP exposure groups. Our data reveal that the EcR pathway changes over MEHP exposure could be associated with lipid accumulation, owing to increased lipid levels and the subsequent increase in the reproduction of MEHP-exposed D. magna.


Asunto(s)
Dietilhexil Ftalato , Animales , Daphnia/metabolismo , Dietilhexil Ftalato/análogos & derivados , Dietilhexil Ftalato/metabolismo , Dietilhexil Ftalato/toxicidad , Femenino , Lípidos , Ácidos Ftálicos , Reproducción , Vitelogeninas
9.
Food Chem Toxicol ; 161: 112829, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-35093429

RESUMEN

Particulate matter (PM) generally consists of aggregated particles containing trace metals and polycyclic aromatic hydrocarbons (PAHs). Cytochrome P450 (CYP) 1A1, one of the extensively investigated biomarkers, is highly inducible when PAHs activate the aryl hydrocarbon receptor (AhR). The present study focused on developing a LC-MS/MS-based assay to evaluate CYP1A1 induction potential following PM exposure. This assay adapted a CYP1A1 selective reaction of granisetron 7-hydroxylation in response to an AhR inducer, 6-formylindolo[3,2-b]carbazole (FICZ), in HepaRG and A549 cell lines. Exposure to FICZ (10 nM) increased the levels of granisetron 7-hydroxylation significantly, whereas no elevation of ethoxyresorufin-O-deethylation (EROD) activity was found in HepaRG cells. In A549 cells, granisetron 7-hydroxylation showed a better dose-response from 0 to 10000 nM FICZ treatment than EROD. EROD Additionally, the application of the assay with diesel PM exposure showed a concentration-dependent induction of CYP1A1 in HepaRG, A549, and human nasal epithelial cells. The granisetron assay has better selectivity for CYP1A1 than the conventional EROD assay, which is overlapped reaction with CYP1A2 and CYP1B1, with high correlations between AhR activation and CYP1A1 mRNA levels. Accompanying the great application potential to different organs and cell culture systems, future studies will implement the granisetron assay for the respiratory toxicity evaluation.


Asunto(s)
Cromatografía Liquida , Citocromo P-450 CYP1A1/metabolismo , Gasolina/análisis , Granisetrón/farmacología , Espectrometría de Masas , Material Particulado/toxicidad , Línea Celular , Citocromo P-450 CYP1A1/genética , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Hepatocitos/efectos de los fármacos , Humanos , Hidroxilación , Material Particulado/química , Alveolos Pulmonares/efectos de los fármacos , ARN Mensajero/genética , ARN Mensajero/metabolismo
10.
Molecules ; 26(4)2021 Feb 08.
Artículo en Inglés | MEDLINE | ID: mdl-33567691

RESUMEN

Steroid 5-α reductase (5AR) is responsible for the reduction of steroids to 5-α reduced metabolites, such as the reduction of testosterone to 5-α dihydrotestosterone (DHT). A new adverse outcome pathway (AOP) for 5AR inhibition to reduce female reproduction in fish (AOP 289) is under development to clarify the antiestrogenic effects of 5AR inhibitors in female fish. A sensitive method for the DHT analysis using chemical derivatization and liquid chromatography-tandem mass spectrometry was developed. A cell-based 5AR inhibition assay that utilizes human cell lines, a transient overexpression system, and fish cell lines was developed. The measured IC50 values of two well-known 5AR inhibitors, finasteride and dutasteride, were comparable in the different systems. However, the IC50 of dutasteride in the fish cell lines was lower than that in the human cell lines. Finasteride showed a higher IC50 against the RTG-2 cell line. These results demonstrated that 5ARs inhibition could differ in terms of structural characteristics among species. The assay has high sensitivity and reproducibility and is suitable for the application in 5AR inhibition screening for various endocrine disruption chemicals (EDCs). Future studies will continue to evaluate the quantitative inhibition of 5AR by EDCs to compare the endocrine-disrupting pathway in different species.


Asunto(s)
3-Oxo-5-alfa-Esteroide 4-Deshidrogenasa/metabolismo , Inhibidores de 5-alfa-Reductasa/farmacología , Cromatografía Liquida , Evaluación Preclínica de Medicamentos/métodos , Espectrometría de Masas , Animales , Calibración , Línea Celular , Humanos , Oncorhynchus mykiss , Pez Cebra
11.
Environ Toxicol Pharmacol ; 64: 187-195, 2018 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-30448727

RESUMEN

Herein, the toxicity of particles generated from the complete combustion of 1 g coal at 500, 700, and 900 °C were compared, and combustion at 700 °C generated the most toxins. Chemical analyses revealed that all components except catechol, resorcinol, and aromatic amines were most abundant at 700 °C. Toxicity results confirmed that the relative mutagenicity, cytotoxicity, redox cycling, and production of reactive oxygen species was highest for particles generated at 700 °C. Particles generated during combustion at 700 °C exhibited higher toxicity toward biological systems due to a higher content of toxic compounds.


Asunto(s)
Contaminantes Atmosféricos/toxicidad , Carbón Mineral , Mutágenos/toxicidad , Material Particulado/toxicidad , Células A549 , Aminas/toxicidad , Supervivencia Celular/efectos de los fármacos , Humanos , Hidrocarburos Aromáticos/toxicidad , Centrales Eléctricas , Especies Reactivas de Oxígeno/metabolismo , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genética , Temperatura
12.
Toxicol Ind Health ; 34(2): 99-109, 2018 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-29415641

RESUMEN

Ultrafine particles (UFPs, < 2.5 µm) in air pollutants have been identified as a major cause of respiratory diseases, since they can affect the lung alveoli through the bronchus. In particular, if toxicants such as heavy metals and polycyclic aromatic hydrocarbons (PAHs) are present in UFPs, they can cause diseases such as asthma, chronic obstructive pulmonary disease, and lung cancer. This study compared in vitro toxicity of various particulate matter including UFPs from combustion particles of diesel (diesel exhaust particles (DEP)), rice straw (RS), pine stem (PS) and coal (CC), and road dust particles from tunnel (TD) and roadside (RD). UFPs from combustion particles and road dust were collected with a glass fiber filter using burning systems and a solid aerosol generator. Cell viability was determined by neutral red uptake assay using Chinese hamster ovary strain K1 cells. Redox cycling activity and intracellular reactive oxygen species were measured using 1,4-dithiothreitol (DTT) and 2',7'-dichlorofluorescin diacetate (DCF-DA) assay, respectively. Our in vitro studies validated that combustion particles had high toxicological activity. PS demonstrated the highest activity in cytotoxicity but DEP had the highest activity in the DTT and DCF-DA assays. Overall, since the toxicological activity of particles generated by various means was different, risk assessment should be conducted through various toxicity evaluations rather than one toxicity evaluation.


Asunto(s)
Contaminantes Atmosféricos/toxicidad , Estrés Oxidativo/efectos de los fármacos , Material Particulado/toxicidad , Mucosa Respiratoria/efectos de los fármacos , Células A549 , Contaminantes Atmosféricos/química , Animales , Células CHO , Supervivencia Celular/efectos de los fármacos , Carbón Mineral/toxicidad , Cricetulus , Polvo , Incendios , Gasolina/toxicidad , Humanos , Dosificación Letal Mediana , Oryza/química , Oxidación-Reducción , Tamaño de la Partícula , Material Particulado/química , Pinus/química , Tallos de la Planta/química , Especies Reactivas de Oxígeno/agonistas , Especies Reactivas de Oxígeno/metabolismo , Mucosa Respiratoria/metabolismo , Emisiones de Vehículos/toxicidad
13.
J Ginseng Res ; 40(3): 292-9, 2016 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-27616906

RESUMEN

BACKGROUND: We previously reported that two-phase partition chromatography between ginseng water extract and soybean oil efficiently eliminated pesticide residues. However, an undesirable odor and an unpalatable taste unique to soybean oil were two major disadvantages of the method. This study was carried out to find an alternative vegetable oil that is cost effective, labor effective, and efficient without leaving an undesirable taste and smell. METHODS: We employed six vegetable oils that were available at a grocery store. A 1-mL sample of the corresponding oil containing a total of 32 pesticides, representing four categories, was mixed with 10% aqueous ginseng extract (20 mL) and equivalent vegetable oil (7 mL) in Falcon tubes. The final concentration of the pesticides in the mixture (28 mL) was adjusted to approximately 2 ppm. In addition, pesticides for spiking were clustered depending on the analytical equipment (GC/HPLC), detection mode (electron capture detector/nitrogen-phosphorus detector), or retention time used. Samples were harvested and subjected to quantitative analysis of the pesticides. RESULTS: Soybean oil demonstrated the highest efficiency in partitioning pesticide residues in the ginseng extract to the oil phase. However, canola oil gave the best result in an organoleptic test due to the lack of undesirable odor and unpalatable taste. Furthermore, the qualitative and quantitative changes of ginsenosides evaluated by TLC and HPLC, respectively, revealed no notable change before or after canola oil treatment. CONCLUSION: We suggest that canola oil is an excellent vehicle with respect to its organoleptic property, cost-effectiveness and efficiency of eliminating pesticide residues in ginseng extract.

14.
J Prosthodont ; 18(1): 43-8, 2009 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-19166547

RESUMEN

PURPOSE: This in vitro study investigated the effect of grooves, remaining tooth structure, and their combination on tooth fracture resistance of endodontically treated anterior teeth with cast dowel and cores. MATERIALS AND METHODS: Sixty extracted maxillary anterior teeth of similar dimensions were endodontically treated and then randomly divided into three groups of 20 teeth each. The teeth in the first group were cut horizontally at the widest part of their anatomical crowns. Three hundred and sixty degree 1 and 2 mm axial walls for ferrule effect were provided for the teeth in the second and the third groups, respectively. Cast dowel and cores were fabricated for all teeth. Each group was then subdivided in two groups: one with no grooves and another with mesial and distal grooves. Hence, six groups were created as follows: (1) teeth with no remaining coronal tooth structure and no grooves (group A-control); (2) teeth with no remaining coronal tooth structure, with mesial and distal grooves (group B); (3) teeth with 1 mm remaining coronal tooth structure, with no grooves (group C); (4) teeth with 1 mm remaining coronal tooth structure, with mesial and distal grooves (group D); (5) teeth with 2 mm remaining coronal tooth structure and no grooves (group E); and (6) teeth with 2 mm remaining coronal tooth structure, with mesial and distal grooves (group F). Complete cast crowns were then fabricated for all teeth. A universal testing machine applied controlled loads to the teeth at a crosshead speed of 2.54 mm/min at an angle of 130 degrees to the long axes of the teeth until failure occurred. The loads were applied 2 mm lower than the incisal edges of the specimens. Descriptive statistics, one-way ANOVA (alpha= 0.05) and Tukey's honestly significant difference (HSD) tests were used to determine the effect of failure loads among the tested groups (alpha= 0.05). RESULTS: The mean failure loads were (N): group A (control), 151.21 +/- 38.18; group B, 221.53 +/- 107.03; group C, 295.35 +/- 81.92; group D, 270.20 +/- 76.01; group E, 491.70 +/- 180.36; group F, 432.67 +/- 193.83. Group E presented the highest failure load and group A (control) the lowest. CONCLUSION: The inclusion of interproximal grooves on the cast dowel and cores of endodontically treated anterior teeth with 1-2 mm of remaining coronal tooth structure does not significantly lower the failure threshold.


Asunto(s)
Análisis del Estrés Dental , Incisivo/fisiopatología , Técnica de Perno Muñón , Fracturas de los Dientes/prevención & control , Preparación Protodóncica del Diente , Diente no Vital/fisiopatología , Cementación , Técnica de Colado Dental , Diseño de Prótesis Dental , Fracaso de la Restauración Dental , Cementos de Ionómero Vítreo , Humanos , Corona del Diente/patología
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