Adaptación marginal de coronas de disilicato de litio obtenidas mediante técnicas de escaneo (CAD/CAM): análisis in vitro con microscopía confocal / Marginal adaptation of lithium disilicate crowns obtained by scanning techniques (CAD/CAM): in vitro confocal microscopy analysis

Objetivo. El objetivo de este estudio fue evaluar la adaptación marginal de coronas de disilicato de litio obtenidas mediante técnicas de escaneo (CAD/CAM), antes y des- pués de la cristalización, a través de análisis in vitro con microscopía confocal (MC). Métodos. Fueron confeccionadas 16 réplicas en poliuretano a partir de la pieza 1.4, de modelo typodont, tallada para corona total. Las réplicas fueron divididas en dos gru- pos, de acuerdo a la técnica de escaneo: Técnica Indirecta (Grupo IND, n=08), donde modelos de yeso fueron escaneados con escáner de laboratorio (inEos X5, Sirona Den- tal Systems) y Técnica Directa (Grupo DIR, n=08), donde modelos typodont fueron escaneados con escáner intraoral (CEREC BlueCam, Sirona Dental Systems). A seguir, se fresaron (inLab MC XL, Sirona Dental Systems) coronas en disilicato de litio (IPS e.max CAD, Ivoclar Vivadent) y se adaptaron a las réplicas. Se evaluó la adaptación marginal con análisis de MC en dos momentos, antes y después de la cristalización del disilicato de litio. Los datos fueron analizados con la prueba de Mann-Whitney, t de Student y Wilcoxon (α= 0,05). Resultados. Hubo una diferencia estadísticamente significativa en la adaptación marginal horizontal entre los grupos IND y DIR después de la cristalización (p=0,05). En el grupo IND, la comparación de la adaptación mar- ginal vertical antes y después de la cristalización mostró una diferencia estadísticamente significativa (p=0,038). Conclusiones. Las coronas de disilicato de litio obtenidas me- diante escaneo directo (CAD/CAM) presentaron menor desajuste marginal vertical. La etapa de cristalización afectó la adaptación marginal de las coronas.

Objective. This study aimed to evaluate lithium disilicate marginal adaption on crowns by scanning techniques (CAD/CAM), before and after crystallization, through confocal microscopy (CM) in vitro analysis. Methods. Sixteen polyurethane replicas were per- formed from tooth 1.4, of a typodont model, prepared for a full crown. The replicas were divided into two groups, according to the scanning technique: Indirect Technique (Group IND, n=08), where dental stone models were scanned with a laboratory scanner (inEos X5, Sirona Dental Systems) and Direct Technique (Group DIR, n=08), where typodont models were scanned with an intraoral scanner (CEREC BlueCam, Sirona Dental Systems). Then, the lithium disilicate crowns (IPS e.max CAD, Ivoclar Vivadent) were milled (inLab MC XL, Sirona Dental Systems) and adapted to the replicas. Margin- al adaptation was evaluated with CM analysis before and after lithium disilicate crystalli- zation. Data were analyzed with the Mann-Whitney, t test, and Wilconxon test (α=0.05). Results. There was a statistically significant difference in horizontal marginal adaptation between IND and DIR groups after crystallization (p=0.05). In IND group, the compar- ison of vertical marginal adaptation before and after crystallization showed a statistically significant difference (p=0.038). Conclusions. Lithium disilicate crowns obtained by direct scanning technique (CAD/CAD) showed less vertical marginal maladjustment. The crystallization stage affected the crown's marginal adaptation.

Pharmacological characterization of the relaxant effect induced by adrenomedullin in rat cavernosal smooth muscle

The aim of the present study was to determine the mechanisms underlying the relaxant effect of adrenomedullin (AM) in rat cavernosal smooth muscle (CSM) and the expression of AM system components in this tissue. Functional assays using standard muscle bath procedures were performed in CSM isolated from male Wistar rats. Protein and mRNA levels of pre-pro-AM, calcitonin receptor-like receptor (CRLR), and Subtypes 1, 2 and 3 of the receptor activity-modifying protein (RAMP) family were assessed by Western immunoblotting and quantitative real-time polymerase chain reaction, respectively. Nitrate and 6-keto-prostaglandin F1α (6-keto-PGF1α; a stable product of prostacyclin) levels were determined using commercially available kits. Protein and mRNA of AM, CRLR, and RAMP 1, -2, and -3 were detected in rat CSM. Immunohistochemical assays demonstrated that AM and CRLR were expressed in rat CSM. AM relaxed CSM strips in a concentration-dependent manner. AM22-52, a selective antagonist for AM receptors, reduced the relaxation induced by AM. Conversely, CGRP8-37, a selective antagonist for calcitonin gene-related peptide receptors, did not affect AM-induced relaxation. Preincubation of CSM strips with NG-nitro-L-arginine-methyl-ester (L-NAME, nitric oxide synthase inhibitor), 1H-(1,2,4)oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, quanylyl cyclase inhibitor), Rp-8-Br-PET-cGMPS (cGMP-dependent protein kinase inhibitor), SC560 [5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3-trifluoromethyl pyrazole, selective cyclooxygenase-1 inhibitor], and 4-aminopyridine (voltage-dependent K+ channel blocker) reduced AM-induced relaxation. On the other hand, 7-nitroindazole (selective neuronal nitric oxide synthase inhibitor), wortmannin (phosphatidylinositol 3-kinase inhibitor), H89 (protein kinase A inhibitor), SQ22536 [9-(tetrahydro-2-furanyl)-9H-purin-6-amine, adenylate cyclase inhibitor], glibenclamide (selective blocker of ATP-sensitive K+ channels), and apamin (Ca2+-activated channel blocker) did not affect AM-induced relaxation. AM increased nitrate levels and 6-keto-PGF1α in rat CSM. The major new contribution of this research is that it demonstrated expression of AM and its receptor in rat CSM. Moreover, we provided evidence that AM-induced relaxation in this tissue is mediated by AM receptors by a mechanism that involves the nitric oxide-cGMP pathway, a vasodilator prostanoid, and the opening of voltage-dependent K+ channels.

Kaurene diterpene induces apoptosis in U87 human malignant glioblastoma cells by suppression of anti-apoptotic signals and activation of cysteine proteases

Gliomas are the most common and malignant primary brain tumors in humans. Studies have shown that classes of kaurene diterpene have anti-tumor activity related to their ability to induce apoptosis. We investigated the response of the human glioblastoma cell line U87 to treatment with ent-kaur-16-en-19-oic acid (kaurenoic acid, KA). We analyzed cell survival and the induction of apoptosis using flow cytometry and annexin V staining. Additionally, the expression of anti-apoptotic (c-FLIP and miR-21) and apoptotic (Fas, caspase-3 and caspase-8) genes was analyzed by relative quantification (real-time PCR) of mRNA levels in U87 cells that were either untreated or treated with KA (30, 50, or 70 µM) for 24, 48, and 72 h. U87 cells treated with KA demonstrated reduced viability, and an increase in annexin V- and annexin V/PI-positive cells was observed. The percentage of apoptotic cells was 9% for control cells, 26% for cells submitted to 48 h of treatment with 50 µM KA, and 31% for cells submitted to 48 h of treatment with 70 µM KA. Similarly, in U87 cells treated with KA for 48 h, we observed an increase in the expression of apoptotic genes (caspase-8, -3) and a decrease in the expression of anti-apoptotic genes (miR-21 and c-FLIP). KA possesses several interesting properties and induces apoptosis through a unique mechanism. Further experiments will be necessary to determine if KA may be used as a lead compound for the development of new chemotherapeutic drugs for the treatment of primary brain tumors.

Morphometric analysis of seromucous acini and granular ducts of submandibular glands from rats, rattus norvegicus, submitted to experimental chronic alcoholism

Ha sido reconocido que la ingesta crónica de etanol causa alteraciones morfológicas en varios tejidos. En el presente trabajo fue realizado un análisis morfométrico de los acinos seromucosos y de los conductos granulosos de las glándulas submandibulares de ratones sometidos a alcoholismo crónico experimental. Ratones Wistar machos fueron sometidos a dieta alcohólica con etanol al 6 por ciento (v/v). A los 5, 10 y 15 meses tras el inicio del tratamiento, fueron recolectadas muestras de las glándulas submandibulares para analizar el área de los ácinos seromucosos y conductos granulosos. Los resultados indicaron que el consumo crónico de etanol reduce significativamente el área de las células de los acinos seromucosos y de las células de los conductos granulosos. Además, hubo un ensanchamiento en el área de los acinos seromucosos y de los conductos granulosos tras el consumo crónico de etanol. Concluimos que los efectos del alcohol fueron más graves mientras mayor fue el periodo de tratamiento