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1.
Int. j. morphol ; 38(5): 1496-1507, oct. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1134467

ABSTRACT

RESUMEN: En la enfermedad hepática crónica el trasplante ortotópico es la única alternativa terapéutica actual pero es limitada por falta de donantes. Ensayos con células madre adultas en daño hepático agudo evidencian promisorios resultados. El objetivo de este trabajo fue evaluar en ratas con daño hepático crónico la efectividad de la infusión de células madre adiposas humanas (CMAd-h). Ratas con fibrosis hepática inducida por tioacetamida fueron agrupadas en: grupo I control que no recibió tioacetamida ni células madre, grupo II recibió tioacetamida y suero fisiológico i.v., grupo III recibió tioacetamida y células madre adiposas 1 x 106/kg i.v. vía vena de la cola. La regeneración hepática histológica se evaluó por el index METAVIR, mientras las Macrophagocytus stellatus, células estrelladas a- SMA+ y células colágeno I+ por inmunohistoquímica; el daño funcional se evaluó por los niveles sanguíneos de los analitos Aspartato Aminotransferasa (AST), Alanina Aminotransferasa (ALT), Fosfatasa Alcalina (ALP), úrea y nitrógeno ureico (BUN) y hemograma. Los resultados muestran atenuación del daño estructural hepático evidenciado por disminución de los nódulos, del grado de lesión histológica en el score Metavir, y disminución de Macrophagocytus stellatus, células a-SMA+ y células colágeno tipo I+; funcionalmente hay reducción moderada de AST, ALT, urea, BUN y disminución moderada de células blancas pero efecto favorable sobre el volumen corpuscular media y la hemoglobina corpuscular media. Ocho semanas después de la infusión hay escasa población de CMAd-h en el hígado. En conclusión la infusión intravenosa de CMAd-h en ratas disminuye el daño funcional y estructural de la fibrosis hepática con escasa persistencia de CMAd-h en el parénquima hepático. A nuestro conocimiento este es el primer trabajo que evalúa el efecto de las CMAd-h en el modelo daño hepático crónico murino y la persistencia de las células trasplantadas.


SUMMARY: In chronic liver disease, orthotopic transplantation is the only current therapeutic alternative but it is limited due to lack of donors. Trials with adult stem cells in acute liver damage show promising results. The aim of this work was to evaluate the effectiveness of human adipose stem cell (h-ASC) infusion in rats with chronic liver damage. Rats with thioacetamide- induced liver fibrosis were grouped into: group I control that did not receive thioacetamide and h-ASC, group II received thioacetamide and saline i.v., group III received thioacetamide and h-ASC 1 x 106/ kg i.v. via tail vein. Histological liver regeneration was evaluated by METAVIR index, while Macrophagocytus stellatus (Kupffer cells), stellate cells a-SMA+ and collagen I+ cells by immunohistochemistry; functional damage was evaluated by blood levels of the analytes Aspartate Aminotransferase (AST), Alanine Aminotransferase (ALT), Alkaline Phosphatase (ALP), Urea and Blood Urea Nitrogen (BUN) and hemogram. The results show attenuation of structural liver damage evidenced by decreased nodules, degree of histologic injury on Metavir score, and decreased Macrophagocytus stellatus, a-SMA+ cells and type I+ collagen cells; functionally there is moderate reduction of AST, ALT, urea, BUN and moderate decrease of white cells but favorable effect on mean corpuscular volume and mean corpuscular hemoglobin. Eight weeks after infusion there is a small population of h-ASC in the liver. In conclusion, intravenous infusion of h-ASC in rats reduces functional and structural damage of hepatic fibrosis with low persistence of h- ASC in the liver parenchyma. To our knowledge this is the first work that evaluates the effect of h-SC in the model of chronic murine liver damage and the persistence of transplanted cells.


Subject(s)
Animals , Female , Rats , Mesenchymal Stem Cell Transplantation/methods , Liver Cirrhosis, Experimental/therapy , Aspartate Aminotransferases/analysis , Immunohistochemistry , Treatment Outcome , Alanine Transaminase/analysis , Disease Models, Animal , Alkaline Phosphatase/analysis , Cell- and Tissue-Based Therapy/methods , Liver Cirrhosis, Experimental/pathology
2.
Rev. chil. obstet. ginecol. (En línea) ; 85(supl.1): S101-S105, set. 2020. tab
Article in Spanish | LILACS | ID: biblio-1138654

ABSTRACT

INTRODUCCIÓN: Las alteraciones del perfil hepático durante el embarazo ocurren en 3-5% de las gestantes. Una nueva etiología que se ha presentado en el contexto de pandemia actual es el síndrome respiratorio agudo severo relacionado con el nuevo coronavirus (SARS-CoV-2). Éste es responsable de alteraciones hepáticas en 2 a 11% de la población general infectada por este virus, y de hasta un 30% en las embarazadas que se infectan con SARS-CoV-2. Con el objetivo de mostrar una presentación poco frecuente del SARS-CoV-2 se expone un caso clínico de elevación de transaminasas en embarazada inducida por este nuevo virus. CASO CLÍNICO: Paciente de 36 años, cursando embarazo de 20+6 semanas, consulta por dolor abdominal asociado a ictericia y coluria. Se solicita estudio donde destaca elevación de transaminasas. Ecografía abdominal con vía biliar fina. Se descartan diferentes etiologías de hepatitis aguda y crónica (dada la falta de antecedentes). Finalmente se solicita PCR para COVID-19 que resulta positiva. CONCLUSIÓN: Luego de un estudio exhaustivo de diferentes etiologías de elevación de transaminasas, se atribuye esta alteración enzimática a SARS-CoV-2. Se decide seguimiento ambulatorio estricto con pruebas hepáticas cada dos semanas. La paciente evoluciona estable con exámenes normales luego de un mes desde que se indica el alta hospitalaria. Después de descartar etiologías frecuentes de elevación de transaminasas durante el embarazo, sugerimos solicitar el estudio de este virus con PCR para COVID-19, ya que podría ser una presentación poco frecuente de SARS-CoV-2.


INTRODUCTION: Approximately 3-5% of women present alterations of hepatic enzymes during pregnancy. Under the new circumstances that the world is facing with the SARS-COV2 pandemic, a new etiology for hepatic enzyme alterations has risen. The severe acute respiratory syndrome that the novel coronavirus causes is responsible for hepatic enzyme alterations in 2 to 11% of the sick population that did not have a previous underlying hepatic condition. Furthermore, hepatic enzyme alterations in pregnant women infected with SARS-COV2 presents in up to 30% of the cases. An infrequent presentation of SARS-COV2 is presented as our clinical case. CLINICAL CASE: A 36-year-old patient with a 20+6 week pregnancy presents abdominal pain, jaundice and choluria. General blood workup shows elevated transaminases. The abdominal ultrasound revealed a thin bile duct. Acute and chronic hepatitis etiologies were discarded. Finally, a PCR of COVID-19 was solicited, which turned out to be positive. CONCLUSIÓN: After an exhaustive study to determine the etiology of the elevated transaminases, the hepatic alterations were attributed to SARS-COV2 infection. A conservative management was adopted, with outpatient follow-up with liver testing every two weeks. The patient progresses with a stable steady decline in hepatic enzyme levels, and one-month post hospital discharge, her transaminases had reached normal values. Based on this clinical case, after ruling out frequent etiologies for elevated transaminases during pregnancy, it seems reasonable to request a PCR for COVID-19, since it could be a rare presentation of SARS-CoV-2.


Subject(s)
Humans , Female , Pregnancy , Adult , Pneumonia, Viral/complications , Pregnancy Complications, Infectious/enzymology , Pregnancy Complications, Infectious/etiology , Coronavirus Infections/complications , Betacoronavirus , Pneumonia, Viral/enzymology , Transferases/analysis , Coronavirus Infections/enzymology , Alkaline Phosphatase/analysis , Pandemics , Jaundice , Liver Diseases/enzymology , Liver Diseases/etiology
3.
Int. j. morphol ; 38(1): 61-68, Feb. 2020. tab, graf
Article in English | LILACS | ID: biblio-1056398

ABSTRACT

Fruit purees can be added to diet as alternative sources of bioactive compounds for the prevention and/or improvement of the complications of metabolic syndrome. In this work we evaluated the effect of the intake of low-fat diets enriched with fruit purees (guava-strawberry, guava-blackberry, guava-soursop, guava-passion fruit) on the body weight and biochemical markers in metabolic syndrome analogy (MSA)-induced rats. The rats (n=6 for each treatment) were induced with a high fat diet and were injected with streptozotocin, one dose every week for 4 consecutive weeks after fasting overnight, then healthy rats were fed with standard diet and MS rats were fed with standard diet plus each of the fruit puree, for 4 weeks. As novel findings, the diet enriched with fruit purees was associated with a reduction in body weight (~13-21 %) and a control in the metabolism of glucose by decreasing plasma glucose (~5963 %). Also, there was a reduction in the total cholesterol, triacylglycerols, low-density lipoproteins, and low enzymatic activities of alanine aminotransferase, alkaline phosphatase and γ-glutamyl transferase, useful metabolites in the control of inflammatory processes in the liver. A notable improvement in the liver morphology was observed indicating that the treatments had a hepatoprotective effect. The diet enriched with guava-blackberry puree caused the best results on most biochemical markers of MS rats. Therefore, diets enriched with fruit purees can be an alternative for MS individuals for the control and improvement of the complications caused by this syndrome.


Los purés de frutas se pueden agregar a la dieta como fuentes alternativas de compuestos bioactivos para la prevención y / o mejora de las complicaciones del síndrome metabólico. En este trabajo evaluamos el efecto de la ingesta de dietas bajas en grasas, enriquecidas con purés de frutas (guayaba-fresa, guayaba-mora, guayaba-guanábana, guayaba-maracuyá) sobre el peso corporal y los marcadores bioquímicos en el síndrome metabólico (SM) inducido en ratas. Las ratas (n = 6 para cada tratamiento) fueron inducidas con una dieta alta en grasas y se les inyectó estreptozotocina, una dosis cada semana durante 4 semanas consecutivas después de ayunar durante la noche. Luego, las ratas sanas fueron alimentadas con una dieta estándar; y las ratas con SM fueron alimentadas con dieta estándar más cada uno de los purés de frutas, durante 4 semanas. Como hallazgos novedosos, la dieta enriquecida con purés de frutas se asoció con una reducción en el peso corporal (~ 13-21 %) y un control en el metabolismo de la glucosa al disminuir la glucosa en plasma (~ 59-63 %). Además, hubo una reducción en el colesterol total, triacilgliceroles, lipoproteínas de baja densidad, y bajas actividades enzimáticas de alanina aminotransferasa, fosfatasa alcalina y gama-glutamil transferasa, metabolitos útiles en el control de los procesos inflamatorios en el hígado. Se observó una mejora notable en la morfología del hígado, lo que indica que los tratamientos tuvieron un efecto hepatoprotector. La dieta enriquecida con puré de guayaba y mora causó los mejores resultados en la mayoría de los marcadores bioquímicos de las ratas con SM. Por lo tanto, las dietas enriquecidas con purés de frutas pueden ser una alternativa para las personas con SM, para el control y la mejora de las complicaciones causadas por este síndrome.


Subject(s)
Animals , Rats , Diet, Fat-Restricted , Metabolic Syndrome , Fruit , Liver/drug effects , Blood Glucose/drug effects , Body Weight/drug effects , Biomarkers , Albumins/analysis , Disease Models, Animal , Alkaline Phosphatase/analysis , Hepatoprotector Drugs , Transaminases/analysis , Lipids/analysis , Liver/chemistry
4.
J. appl. oral sci ; 28: e20190105, 2020. tab, graf
Article in English | LILACS | ID: biblio-1056578

ABSTRACT

Abstract Calcium aluminate cement (CAC) has been highlighted as a promising alternative for endodontic use aiming at periapical tissue repair. However, its effects on dental pulp cells have been poorly explored. Objective: This study assessed the impact of calcium chloride (CaCl2) and bismuth oxide (Bi2O3) or zinc oxide (ZnO) additives on odontoblast cell response to CAC. Methodology: MDPC-23 cells were exposed for up to 14 d: 1) CAC with 2.8% CaCl2 and 25% ZnO (CACz); 2) CAC with 2.8% CaCl2 and 25% Bi2O3 (CACb); 3) CAC with 10% CaCl2 and 25% Bi2O3 (CACb+); or 4) mineral trioxide aggregate (MTA), placed on inserts. Non-exposed cultures served as control. Cell morphology, cell viability, gene expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), and dentin matrix protein 1 (DMP-1), ALP activity, and extracellular matrix mineralization were evaluated. Data were compared using ANOVA (α=5%). Results: Lower cell density was detected only for MTA and CACb+ compared with Control, with areas showing reduced cell spreading. Cell viability was similar among groups at days one and three (p>0.05). CACb+ and MTA showed the lowest cell viability values at day seven (p>0.05). CACb and CACb+ promoted higher ALP and BSP expression compared with CACz (p<0.05); despite that, all cements supported ALP activity. Matrix mineralization were enhanced in CACb+ and MTA. Conclusion: In conclusion, CAC with Bi2O3, but not with ZnO, supported the expression of odontoblastic phenotype, but only the composition with 10% CaCl2 promoted mineralized matrix formation, rendering it suitable for dentin-pulp complex repair.


Subject(s)
Humans , Mice , Calcium Compounds/pharmacology , Calcium Compounds/chemistry , Aluminum Compounds/pharmacology , Aluminum Compounds/chemistry , Dental Cements/pharmacology , Dental Cements/chemistry , Dental Pulp/cytology , Dental Pulp/drug effects , Oxides/pharmacology , Oxides/chemistry , Time Factors , Zinc Oxide/pharmacology , Zinc Oxide/chemistry , Bismuth/pharmacology , Bismuth/chemistry , Materials Testing , Calcium Chloride/pharmacology , Calcium Chloride/chemistry , Gene Expression/drug effects , Cell Survival/drug effects , Cells, Cultured , Reproducibility of Results , Silicates/pharmacology , Silicates/chemistry , Drug Combinations , Alkaline Phosphatase/analysis , Alkaline Phosphatase/drug effects , Odontoblasts/drug effects
5.
Adv Rheumatol ; 60: 15, 2020. tab
Article in English | LILACS | ID: biblio-1088639

ABSTRACT

Abstract Background: Bone disease is common in patients undergoing hemodialysis. It is the result of bone turnover abnormalities and the decrease of bone mineral density (BMD). We aimed to determine the usefulness of serum bone turnover markers and BMD measurement by dual-energy x-ray absorptiometry (DXA) in hemodialysis patients. Methods: We conducted a cross-sectional study including 90 hemodialysis for more than 12 months. Bone mineral density was assessed by DXA. Peripheral blood samples were obtained from each patient before dialysis in a fasting state within a week of the DXA. Biochemical variables of calcium and phosphate were measured. One bone formation marker (bone-specific alkaline phosphatase (bAP), one bone resorption marker (carboxy-terminal telopeptides of type 1 collagen (CTX)) were measured. Total alkaline phosphatase (TAP), intact parathyroid hormone (PTH) and fibroblast growth factor 23 (FGF23) which is a bone-derived hormone were also measured. Results: CTX values were 6.25 times higher than the normal limit of the assay. Bone alkaline phosphatase levels were less than 10 ng/mL in 28.8% of cases. 23% of patients have osteoporosis and 45% have osteopenia. Femoral BMD had negative correlations with age and PTH levels. FGF23 levels were significantly increased in patients with osteoporosis affecting the lumbar. The levels of bAP and CTX showed a positive correlation. Both circulating bAP and CTX levels showed also positive correlations with PTH levels. Fractures, observed in 12.2% of cases, were associated with low PTH values and the existence of osteoporosis. Conclusions: Our study showed that osteoporosis and fracture are common in dialysis patients. The reduced BMD was associated with advanced age and elevated levels of PTH. Markers of bone turnover and FGF23 may play a role in the diagnosis of bone disease in hemodialysis patients. DXA measurement is necessary for the monitoring for bone loss.(AU)


Subject(s)
Humans , Osteoporosis/diagnosis , Bone Density , Renal Dialysis/adverse effects , Bone Resorption , Cross-Sectional Studies/instrumentation , Collagen Type I/analysis , Alkaline Phosphatase/analysis , Fibroblast Growth Factors/analysis
6.
J. pediatr. (Rio J.) ; 95(5): 567-574, Sept.-Oct. 2019. tab
Article in English | LILACS | ID: biblio-1040356

ABSTRACT

Abstract Objective: The objective of this study was to evaluate the effects of two low-dose combined oral contraceptives on bone metabolism in adolescents for one year. Methods: This was a quasi-experimental study. The adolescents were divided into three groups: oral contraceptives 1 (n = 42) (20 µg EE/150 µg desogestrel), oral contraceptives 2 (n = 66) (30 µg EE/3 mg drospirenone), and a control group (n = 70). Adolescents underwent anthropometric assessment and densitometry (dual-energy X-ray). Bone age and bone formation markers (osteocalcin and bone alkaline phosphatase) were evaluated. The oral contraceptives users were evaluated again after 12 months. Linear regression analysis was used to indirectly study the effect of each additional year of chronological age on anthropometric and densitometric variables as well as on bone markers in the control group. Results: At study entry, no significant differences were observed between the oral contraceptives 1, oral contraceptives 2, and controls in the analyzed variables. Linear regression analysis showed an increase in bone mineral density and bone mineral content for each additional year. There was a significant reduction in bone alkaline phosphatase levels; no significant difference was observed for osteocalcin in control individuals. Comparison of dual-energy X-ray variables at baseline and after one year showed no significant differences in the oral contraceptives 1 or oral contraceptives 2 groups. A significant reduction in bone alkaline phosphatase and osteocalcin levels was observed in both the oral contraceptives 1 and oral contraceptives 2 groups. Conclusion: Adolescent women gain peak bone mass during this phase of life. Two low-dose combined oral hormonal contraceptives were associated with lower bone gain and lower bone formation markers than in untreated controls.


Resumo: Objetivo: O objetivo deste estudo foi avaliar os efeitos de dois contraceptivos orais combinados de baixa dosagem por um ano sobre o metabolismo ósseo em adolescentes. Métodos: Este foi um estudo quase experimental. As adolescentes foram divididas em três grupos: contraceptivos orais 1 (n = 42) (20 µg de EE/150 µg de desogestrel), contraceptivos orais 2 (n = 66) (30 µg EE/3 mg de drospirenona) e grupo controle (n = 70). As adolescentes foram submetidas à avaliação antropométrica e densitometria (raio-X de dupla energia). Foram avaliados a idade óssea e os marcadores de formação óssea (osteocalcina e fosfatase alcalina óssea). As usuárias de contraceptivos orais foram novamente avaliadas após 12 meses. A análise de regressão linear foi utilizada para estudar, indiretamente, o efeito de cada ano adicional da idade cronológica sobre as variáveis antropométricas e densitométricas e sobre os marcadores ósseos no grupo de controle. Resultados: No início do estudo, não foram observadas diferenças significativas nas variáveis analisadas entre as usuárias de contraceptivos orais 1, contraceptivos orais 2 e o grupo controle. A análise de regressão linear mostrou um aumento na densidade mineral óssea e no conteúdo mineral ósseo para cada ano adicional. Houve uma redução significativa nos níveis de fosfatase alcalina óssea e não foi observada diferença significativa para osteocalcina nos indivíduos controles. A comparação das variáveis do raio-X de dupla energia no início e após um ano não mostrou diferença significativa no grupo de contraceptivos orais 1 ou contraceptivos orais 2. Foi observada uma redução significativa nos níveis de fosfatase alcalina óssea e osteocalcina nos dois grupos contraceptivos orais 1 e contraceptivos orais 2. Conclusão: As adolescentes atingiram o pico de massa óssea durante essa fase da vida. Duas formulações de contraceptivos hormonais orais de baixa dosagem, após um ano de uso, se associaram a menor incremento na densidade mineral óssea e menor concentração de marcadores de formação óssea quando confrontados com resultados de adolescentes não usuárias de contraceptivos.


Subject(s)
Humans , Female , Child , Adolescent , Young Adult , Osteogenesis/drug effects , Bone Density/drug effects , Desogestrel/administration & dosage , Contraceptives, Oral, Hormonal/administration & dosage , Ethinyl Estradiol/administration & dosage , Androstenes/administration & dosage , Osteogenesis/physiology , Reference Values , Time Factors , Bone Density/physiology , Linear Models , Osteocalcin/analysis , Anthropometry , Analysis of Variance , Statistics, Nonparametric , Alkaline Phosphatase/analysis , Non-Randomized Controlled Trials as Topic
7.
Int. j. morphol ; 37(1): 237-240, 2019. tab, graf
Article in English | LILACS | ID: biblio-990033

ABSTRACT

SUMMARY: Brassica juncea (Indian mustard) seeds are consumed in treatment of high blood pressure, headache and prevention of heart disease. The aim of the present study was to investigate the effects of methanol extract of Brassica juncea seeds [BJME] on the heart and liver of adult Albino Wistar rats. A total of 24 albino rats of both sexes were divided into 6 groups [I - VI] of 4 rats per group. Groups I to IV received graded doses of the methanol extract by oral gavage while groups V and VI (controls) received 2 ml/kg body weight of 3 % Tween 80 and water respectively via oral gavage once daily. Treatment lasted for four weeks and the serum levels of aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (ALP) were estimated. The animals were sacrificed and the heart and liver tissues were excised for further histological processing for light microscopy. There was significant increase in AST and ALT levels following BJME treatment when compared to the controls. ALP activity did not differ significantly among the treatment and control groups. Histopathological changes consistent with toxic injury were observed in the heart and liver tissues of BJME- treated rats. In conclusion, the results of this study show that sub-acute administration of methanol seed extract of Brassica juncea can exert cardiotoxic and hepatotoxic effects in rats.


RESUMEN: Las semillas de Brassica juncea (mostaza india) se consumen en el tratamiento de la hipertensión arterial, el dolor de cabeza y la prevención de enfermedades del corazón. El objetivo del presente estudio fue investigar los efectos del extracto de metanol de semillas de Brassica juncea [BJME] en el corazón y el hígado de ratas Albino Wistar adultas. Un total de 24 ratas albinas de ambos sexos se dividieron en 6 grupos [I - VI] de 4 ratas por grupo. Los grupos I a IV recibieron dosis del extracto de metanol por sonda oral progresivamente, mientras que los grupos V y VI (control) recibieron 2 ml / kg de peso corporal de 3 % de 80 y agua, respectivamente, por sonda oral una vez al día. El tratamiento duró cuatro semanas y se estimaronlos niveles séricos de aspartato transaminasa (AST), alanina transaminasa (ALT) y fosfatasa alcalina (ALP). Los animales se sacrificaron y fueron analizados los tejidos del corazón y el hígado, para un procesamiento histológico adicional con microscopía óptica. Hubo un aumento significativo en los niveles de AST y ALT después del tratamiento con BJME en comparación con los controles. La actividad de ALP no difirió significativamente entre los grupos de tratamiento y control. Se observaron cambios histopatológicos compatibles con lesiones tóxicas en los tejidos del corazón y el hígado de ratas tratadas con BJME. En conclusión, los resultados de este estudio muestran que la administración subaguda de extracto de semilla de metanol de Brassica juncea puede ejercer efectos cardiotóxicos y hepatotóxicos en ratas.


Subject(s)
Animals , Rats , Plant Extracts/pharmacology , Methanol/pharmacology , Heart/drug effects , Liver/drug effects , Mustard Plant/chemistry , Aspartate Aminotransferases/analysis , Seeds , Time Factors , Plant Extracts/administration & dosage , Rats, Wistar , Alanine Transaminase/analysis , Methanol/administration & dosage , Alkaline Phosphatase/analysis
8.
J. appl. oral sci ; 27: e20180429, 2019. tab, graf
Article in English | LILACS, BBO | ID: biblio-990101

ABSTRACT

Abstract Objectives: To evaluate the radiopacity of Biodentine (BD) and BD associated with 15% calcium tungstate (BDCaWO4) or zirconium oxide (BDZrO2), by using conventional and digital radiography systems, and their physicochemical and biological properties. Materials and Methods: Radiopacity was evaluated by taking radiographs of cement specimens (n=8) using occlusal film, photostimulable phosphor plates or digital sensors. Solubility, setting time, pH, cytocompatibility and osteogenic potential were also evaluated. Data were analyzed using one-way ANOVA and Tukey post-test or two-way ANOVA and Bonferroni post-test (α=0.05). Results: BD radiopacity was lower than 3 mm Al, while BD ZrO2 and BD CaWO4 radiopacity was higher than 3 mm Al in all radiography systems. The cements showed low solubility, except for BDCaWO4. All cements showed alkaline pH and setting time lower than 34 minutes. MTT and NR assays revealed that cements had greater or similar cytocompatibility in comparison with control. The ALP activity in all groups was similar or greater than the control. All cements induced greater production of mineralized nodules than control. Conclusions: Addition of 15% ZrO2 or CaWO4 was sufficient to increase the radiopacity of BD to values higher than 3 mm Al. BD associated with radiopacifiers showed suitable properties of setting time, pH and solubility, except for BDCaWO4, which showed the highest solubility. All cements had cytocompatibility and potential to induce mineralization in Saos-2 cells. The results showed that adding 15% ZrO2 increases the radiopacity of BD, allowing its radiography detection without altering its physicochemical and biological properties.


Subject(s)
Humans , Zirconium/chemistry , Tungsten Compounds/chemistry , Silicates/chemistry , Calcium Compounds/chemistry , Radiography, Dental, Digital/methods , Osteoblasts/drug effects , Reference Values , Solubility , Time Factors , Zirconium/pharmacology , Materials Testing , Cell Survival/drug effects , Reproducibility of Results , Analysis of Variance , Anthraquinones , Tungsten Compounds/pharmacology , Silicates/pharmacology , Calcium Compounds/pharmacology , Alkaline Phosphatase/analysis , Hydrogen-Ion Concentration
9.
J. appl. oral sci ; 27: e20180317, 2019. tab, graf
Article in English | LILACS, BBO | ID: biblio-984571

ABSTRACT

Abstract Bone morphogenetic protein type 2 (BMP-2) and retinoic acid (RA) are osteoinductive factors that stimulate endogenous mechanisms of bone repair which can be applied on management of osseous defects in oral and maxillofacial fields. Objective Considering the different results of RA on osteogenesis and its possible use to substitute/potency BMP-2 effects, this study evaluated the outcomes of BMP-2, RA, and BMP-2+RA treatments on in vitro osteogenic differentiation of human adipose-derived stem cells (ASCs) and the signaling pathway(s) involved. Material and Methods ASCs were treated every other day with basic osteogenic medium (OM) alone or supplemented with BMP-2, RA, or BMP-2+RA. Alkaline phosphatase (ALP) activity was determined using the r-nitrophenol method. Extracellular matrix mineralization was evaluated using von Kossa staining and calcium quantification. Expression of osteonectin and osteocalcin mRNA were determined using qPCR. Smad1, Smad4, phosphorylated Smad1/5/8, BMP-4, and BMP-7 proteins expressions were analyzed using western blotting. Signaling pathway was evaluated using the IPA® software. Results RA promoted the highest ALP activity at days 7, 14, 21, and 28, in comparison to BMP-2 and BMP-2+RA. BMP-2+RA best stimulated phosphorylated Smad1/5/8 protein expression at day 7 and Smad4 expression at days 7, 14, 21, and 28. Osteocalcin and osteonectin mRNA expressions were best stimulated by BMP-2+RA at day 7. Matrix mineralization was most improved by BMP-2+RA at days 12 and 32. Additionally, BMP-2+RA promoted the highest BMP signaling pathway activation at days 7 and 14, and demonstrated more activation of differentiation of bone-forming cells than OM alone. Conclusions In summary, RA increased the effect of BMP-2 on osteogenic differentiation of human ASCs.


Subject(s)
Humans , Osteogenesis/drug effects , Tretinoin/pharmacology , Cell Differentiation/drug effects , Bone Morphogenetic Protein 2/drug effects , Mesenchymal Stem Cells/drug effects , Osteoblasts/drug effects , Osteogenesis/physiology , Reference Values , Time Factors , Osteocalcin/analysis , Osteocalcin/drug effects , Osteonectin/analysis , Osteonectin/drug effects , Cell Differentiation/physiology , Cells, Cultured , Blotting, Western , Reproducibility of Results , Analysis of Variance , Alkaline Phosphatase/analysis , Alkaline Phosphatase/adverse effects , Bone Morphogenetic Protein 2/metabolism , Mesenchymal Stem Cells/metabolism
10.
Arch. endocrinol. metab. (Online) ; 62(4): 438-445, July-Aug. 2018. tab, graf
Article in English | LILACS | ID: biblio-950087

ABSTRACT

ABSTRACT Objective: This study evaluated the effects of combination therapy of curcumin and alendronate on BMD and bone turnover markers in postmenopausal women with osteoporosis. Subjects and methods: In a randomized, double-blind trial study, 60 postmenopausal women were divided into three groups: control, alendronate, and alendronate + curcumin. Each group included 20 patients. Total body, total hip, lumbar spine and femoral neck BMDs were measured by dual-energy X-ray absorptiometry (DXA) at baseline and after 12 months of therapy. Bone turnover markers such as bone-specific alkaline phosphatase (BALP), osteocalcin and C-terminal cross-linking telopeptide of type I collagen (CTx) were measured at the outset and 6 months later. Results: Patients in the control group suffered a significant decrease in BMD and increased bone turnover markers at the end of study. The group treated with only alendronate showed significantly decreased levels of BALP and CTx and increased levels of osteocalcin compared to the control group. The alendronate group also showed significant increases in the total body, total hip, lumbar spine and femoral neck BMDs at the end of study compared to the control group. In the curcumin + alendronate group, BALP and CTx levels decreased and osteocalcin levels increased significantly at the end of study compared to the control and alendronate groups. BMD indexes also increased in four areas significantly at the end of study compared to the control and alendronate groups. Conclusion: The combination of curcumin and alendronate has beneficial effects on BMD and bone turnover markers among postmenopausal women with osteoporosis. Arch Endocrinol Metab. 2018;62(4):438-45


Subject(s)
Humans , Female , Middle Aged , Aged , Bone Density/drug effects , Osteoporosis, Postmenopausal/metabolism , Alendronate/pharmacology , Curcumin/pharmacology , Bone Density Conservation Agents/pharmacology , Peptide Fragments/drug effects , Peptide Fragments/urine , Osteocalcin/analysis , Osteocalcin/drug effects , Double-Blind Method , Bone Remodeling/drug effects , Collagen Type II/drug effects , Collagen Type II/urine , Drug Therapy, Combination/methods , Alkaline Phosphatase/analysis , Alkaline Phosphatase/drug effects
11.
J. appl. oral sci ; 26: e20170470, 2018. graf
Article in English | LILACS, BBO | ID: biblio-954503

ABSTRACT

Abstract The hypothesis of this study was that the peri-implant bone healing of the group of pinealectomized rats would differ from the control group. The samples were subjected to immunohistochemical, microtomographic (total porosity and connectivity density), and fluorochrome (mineralized surface) analyses. Objectives The goal of this study was to investigate the cellular changes and bone remodeling dynamics along the bone/implant interface in pinealectomized rats. Material and Methods The total of 18 adult male rats (Rattus norvegicus albinus, Wistar) was divided into three groups (n=6): control (CO), pinealectomized without melatonin (PNX) and pinealectomized with melatonin (PNXm). All animals were submitted to the first surgery (pinealectomy), except the CO group. Thirty days after the pinealectomy without melatonin, the second surgery was conducted, in which all animals received an implant in each tibia (36 titanium implants with surface treatment were installed - Implalife® São Paulo, SP, Brazil). By gavage, the rats of the PNX group received the vehicle solution, and the procedure. Results Immunohistochemical analysis for runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteopontin (OP) and osteocalcin (OC) showed that the bone repair process in the PNXm group was similar to that of the CO group, whereas the PNX group showed a delay. The microtomographic parameters of total porosity [Po(tot)] and bone surface (BS) showed no statistically significant differences, whereas for the connective density (Conn.Dn) a statistical difference was found between the CO and PNXm groups. Fluorochrome analysis of the active mineralized surface showed statistically significant difference between the CO and PNX and between the CO and PNXm groups. Conclusion The absence of the pineal gland impaired the bone repair process during osseointegration, however the daily melatonin replacement was able to restore this response.


Subject(s)
Animals , Male , Pineal Gland/surgery , Osseointegration/drug effects , Bone Density Conservation Agents/pharmacology , Bone-Implant Interface , Melatonin/pharmacology , Tibia/drug effects , Tibia/injuries , Tibia/pathology , Titanium , Immunohistochemistry , Osteocalcin/analysis , Reproducibility of Results , Treatment Outcome , Rats, Wistar , Implants, Experimental , Dental Implantation, Endosseous , Alkaline Phosphatase/analysis , Core Binding Factor Alpha 1 Subunit/analysis , Osteopontin/analysis , X-Ray Microtomography , Fluorescent Dyes
12.
Rev. Hosp. Ital. B. Aires (2004) ; 37(2): 63-67, jun. 2017. graf., ilus.
Article in Spanish | LILACS | ID: biblio-1087149

ABSTRACT

Presentamos un paciente de 63 años con cáncer renal y aumento de fosfatasa alcalina sérica de tipo óseo de acuerdo con su reactividad con anticuerpos monoclonales específicos. Se descartaron las causas conocidas de aumento de la isoenzima, incluyendo metástasis óseas. Los niveles enzimáticos cayeron abruptamente con la remoción del tumor, por lo que consideramos a este último como su origen. Diversas isoenzimas de fosfatasa alcalina pueden ser producidas y secretadas por tumores como manifestación paraneoplásica. El conocimiento de esto puede, en ocasiones, orientarnos hacia la presencia de una neoplasia oculta. Además, los cambios en los niveles séricos de esas isoenzimas pueden ser indicadores de respuesta al tratamiento o de recidiva tumoral. (AU)


A 63-year old man was seen in the outpatient clinic because of renal cancer and elevation in bone alkaline phosphatase measured by monoclonal antibodies assay. Known causes of bone isoenzyme augmentation, including bone metastases, were ruled out. The tumoral origin of the isoenzyme was diagnosed because after removal of the tumor the enzymatic levels fell sharply. Several alkaline phosphatase isoenzymes can be produced and secreted by tumors as a paraneoplasic manifestation and their elevation could be a manifestation of an occult neoplasia. Furthermore the monitoring of their blood levels can be useful means of treatment response and a tool to monitoring recurrence if a sharp decrease after removal of the tumor is observed. (AU)


Subject(s)
Humans , Male , Middle Aged , Alkaline Phosphatase/biosynthesis , Kidney Neoplasms/metabolism , Osteitis Deformans/diagnostic imaging , Atenolol/therapeutic use , Biomarkers , Erythropoietin/therapeutic use , Simvastatin/therapeutic use , Alkaline Phosphatase/analysis , Alkaline Phosphatase/radiation effects , Alkaline Phosphatase/physiology , Everolimus/therapeutic use , Sunitinib/therapeutic use , Zoledronic Acid/therapeutic use , Hypercholesterolemia/drug therapy , Hypertension/drug therapy , Ilium/diagnostic imaging , Anemia/drug therapy , Kidney Neoplasms/pathology , Kidney Neoplasms/drug therapy , Kidney Neoplasms/diagnostic imaging , Antibodies, Monoclonal/radiation effects
13.
J. appl. oral sci ; 25(3): 299-309, May-June 2017. graf
Article in English | LILACS, BBO | ID: biblio-893619

ABSTRACT

Abstract Objective To assess the effect of fibronectin (Fn) and porcine type I collagen (PCOL) on odontoblast-like cells in vitro. Material and Methods Rat odontoblast-like cells (MDPC-23 cells) were inoculated and cultured on Fn-coated or type I collagen-coated substrates. Proliferation assay, alkaline phosphatase activity (ALP activity), mRNA expression of hard tissue-forming markers, and Alizarin red staining were investigated over a period of 10 days. Results Cells maintained a high proliferation activity on Fn and PCOL even at a low seeding concentration (0.5×104/mL) as demonstrated by CCK-8 assay. The proliferation activity of cells on Fn increases in a concentration-dependent manner while it reached a plateau after 10 µg/mL. Cells adopted long, thin and spindle shape on Fn(10-50) and PCOL. Parallel actin filaments were observed in MDPC-23 cells cultured on Fn and PCOL. ALP activity was markedly up-regulated on Fn and PCOL-coated surfaces. Importantly, gene expression of BSP (Fn10: 2.44±0.32; Fn20: 3.05±0.01; Fn30: 2.90±0.21; Fn40: 2.74±0.30; Fn50: 2.64±0.12; PCOL: 2.20±0.03) and OCN (Fn10: 2.52±0.23; Fn20: 2.28±0.24; Fn30: 2.34±0.21; Fn40: 2.34±0.25; Fn50: 2.20±0.22; PCOL: 1.56±0.16) was significantly enhanced on Fn and PCOL substrates as compared with control; moreover, expression of integrin beta 1 (ITGB1), an ubiquitous cell surface receptor was augmented in Fn(10-50) and PCOL groups simultaneously. In accordance with the ALP activity and gene expression data, calcific deposition in cells grown on Fn(10-50) and PCOL was observed as well. Conclusion Despite the limitation of this study, the findings indicate that a surface coating of Fn enhances the proliferation, differentiation and mineralization of odontoblast-like cells by activation of integrin beta 1 (ITG B1). The promoting effects of Fn on MDPC-23 cells were achieved at a comparatively lower coating concentration than type I collagen (300 µg/mL). Specifically, it is suggested that the optimum coating concentration of Fn to be 10 µg/mL.


Subject(s)
Humans , Animals , Rats , Cell Differentiation/drug effects , Fibronectins/pharmacology , Cell Proliferation/drug effects , Odontoblasts/drug effects , Time Factors , Gene Expression , Cells, Cultured , Reproducibility of Results , Fluorescent Antibody Technique , Anthraquinones , Integrin beta1/pharmacology , Reverse Transcriptase Polymerase Chain Reaction , Collagen Type I/pharmacology , Alkaline Phosphatase/analysis
14.
J. appl. oral sci ; 25(1): 42-52, Jan.-Feb. 2017. graf
Article in English | LILACS, BBO | ID: biblio-841161

ABSTRACT

Abstract Sodium alendronate is a bisphosphonate drug that exerts antiresorptive action and is used to treat osteoporosis. Objective The aim of this study was to evaluate the bone repair process at the bone/implant interface of osteoporotic rats treated with sodium alendronate through the analysis of microtomography, real time polymerase chain reactions and immunohistochemistry (RUNX2 protein, bone sialoprotein (BSP), alkaline phosphatase, osteopontin and osteocalcin). Material and Methods A total of 42 rats were used and divided in to the following experimental groups: CTL: control group (rats submitted to fictitious surgery and fed with a balanced diet), OST: osteoporosis group (rats submitted to a bilateral ovariectomy and fed with a low calcium diet) and ALE: alendronate group (rats submitted to a bilateral ovariectomy, fed with a low calcium diet and treated with sodium alendronate). A surface treated implant was installed in both tibial metaphyses of each rat. Euthanasia of the animals was conducted at 14 (immunhostochemistry) and 42 days (immunohistochemistry, micro CT and PCR). Data were subjected to statistical analysis with a 5% significance level. Results Bone volume (BV) and total pore volume were higher for ALE group (P<0.05). Molecular data for RUNX2 and BSP proteins were significantly expressed in the ALE group (P<0.05), in comparison with the other groups. ALP expression was higher in the CTL group (P<0.05). The immunostaining for RUNX2 and osteopontin was positive in the osteoblastic lineage cells of neoformed bone for the CTL and ALE groups in both periods (14 and 42 days). Alkaline phosphatase presented a lower staining area in the OST group compared to the CTL in both periods and the ALE at 42 days. Conclusion There was a decrease of osteocalcin precipitation at 42 days for the ALE and OST groups. Therefore, treatment with short-term sodium alendronate improved bone repair around the implants installed in the tibia of osteoporotic rats.


Subject(s)
Animals , Female , Osteoporosis/drug therapy , Dental Implants , Osseointegration/drug effects , Alendronate/pharmacology , Bone Density Conservation Agents/pharmacology , Osteoblasts/drug effects , Osteoporosis/physiopathology , Tibia/surgery , Time Factors , Immunohistochemistry , Ovariectomy , Bone Density/drug effects , Osteocalcin/analysis , Osteocalcin/drug effects , Cell Differentiation/drug effects , Reproducibility of Results , Rats, Wistar , Implants, Experimental , Dental Implantation, Endosseous , Alkaline Phosphatase/analysis , Alkaline Phosphatase/drug effects , Core Binding Factor Alpha 1 Subunit/analysis , Core Binding Factor Alpha 1 Subunit/drug effects , Osteopontin/analysis , Osteopontin/drug effects , X-Ray Microtomography , Real-Time Polymerase Chain Reaction
15.
Braz. oral res. (Online) ; 31: e17, 2017. tab, graf
Article in English | LILACS | ID: biblio-839523

ABSTRACT

Abstract Periodontitis develops as a result of a continuous interaction between host cells and subgingival pathogenic bacteria. The periodontium has a limited capacity for regeneration, probably due to changes in periodontal ligament stem cells (PDLSCs) phenotype. The aim of this study was to evaluate the effects of lipopolysaccharides from Porphyromonas gingivalis (PgLPS) on mesenchymal phenotype and osteoblast/cementoblast (O/C) potential of PDLSCs. PDLSCs were assessed for Toll-like receptor 2 (TLR2) expression by immunostaining technique. After, cells were exposed to PgLPS, and the following assays were carried out: (i) cell metabolic activity using MTS; (ii) gene expression for IL-1β, TNF-α and OCT-4 by real-time polymerase chain reaction (RT-qPCR); (iii) flow cytometry for STRO-1 and CD105, and (iv) osteogenic differentiation. PDLSCs were positive for TLR2. PgLPS promoted cell proliferation, produced IL-1β and TNF-α, and did not affect the expression of stem cell markers, STRO-1, CD105 and OCT-4. Under osteogenic condition, PDLSCs exposed to PgLPS showed a similar potential to differentiate toward osteoblast/cementoblast phenotype compared to control group as revealed by mineralized matrix deposition and levels of transcripts for RUNX2, ALP and OCN. These results provide evidence that PgLPS induces pro-inflammatory cytokines, but does not change the mesenchymal phenotype and osteoblast/cementoblast differentiation potential of PDLSCs.


Subject(s)
Humans , Osteogenesis/drug effects , Periodontal Ligament/cytology , Lipopolysaccharides/toxicity , Porphyromonas gingivalis , Mesenchymal Stem Cells/drug effects , Time Factors , Gene Expression , Osteocalcin/analysis , Cell Differentiation/drug effects , Cell Survival/drug effects , Cells, Cultured , Tumor Necrosis Factor-alpha/analysis , Statistics, Nonparametric , Cell Proliferation/drug effects , Alkaline Phosphatase/analysis , Octamer Transcription Factor-3/analysis , Toll-Like Receptors/analysis , Core Binding Factor Alpha 1 Subunit/analysis , Interleukin-1beta/analysis , Mesenchymal Stem Cells/metabolism , Real-Time Polymerase Chain Reaction , Flow Cytometry
16.
J. appl. oral sci ; 24(4): 376-382, July-Aug. 2016. graf
Article in English | LILACS, BBO | ID: lil-792596

ABSTRACT

ABSTRACT Aging negatively affects bone/titanium implant interactions. Our hypothesis is that the unbalance between osteogenesis and adipogenesis induced by aging may be involved in this phenomenon. Objective We investigated the osteoblast and adipocyte differentiation of mesenchymal stem cells (MSCs) from young and aged rats cultured on Ti. Material and Methods Bone marrow MSCs derived from 1-month and 21-month rats were cultured on Ti discs under osteogenic conditions for periods of up to 21 days and osteoblast and adipocyte markers were evaluated. Results Cell proliferation, alkaline phosphatase (ALP) activity, extracellular matrix mineralization and gene expression of RUNX2, osterix, ALP, bone sialoprotein, osteopontin, and osteocalcin were reduced in cultures of 21-month rats compared with 1-month rats grown on Ti. Gene expression of PPAR-γ , adipocyte protein 2, and resistin and lipid accumulation were increased in cultures of 21-month rats compared with 1-month rats grown on the same conditions. Conclusions These results indicate that the lower osteogenic potential of MSCs derived from aged rats compared with young rats goes along with the higher adipogenic potential in cultures grown on Ti surface. This unbalance between osteoblast and adipocyte differentiation should be considered in dental implant therapy to the elderly population.


Subject(s)
Animals , Female , Rats , Osteoblasts/physiology , Titanium/chemistry , Aging/physiology , Dental Implants , Adipogenesis/physiology , Mesenchymal Stem Cells/physiology , Osteogenesis/physiology , Surface Properties , Gene Expression , Cells, Cultured , Age Factors , Cell Proliferation/physiology , Alkaline Phosphatase/analysis , Real-Time Polymerase Chain Reaction , Lipids/analysis
17.
Rev. Soc. Odontol. La Plata ; 26(52): 19-21, jun. 2016.
Article in Spanish | LILACS | ID: lil-795818

ABSTRACT

La Fosfatasa Alcalina Ósea (FAO) es una isoforma de la Fosfatasa Alcalina (FAL). La medición de su actividad en saliva es una medida indirecta del proceso de formación ósea, más sensible y específica que la FAL. La catepsina K es la principal colagenasa del proceso de resorción ósea, es capaz de degradar al colágeno tipo I en varios sitios dando lugar a pequeños péptidos N- y C- terminales. El telopéptido C-terminal (CTx) es el marcador más sensible y específico en el aumento de la resorción ósea, ya que el colágeno tipo I constituye más del 90 por ciento de la matriz orgánica del hueso...


Subject(s)
Humans , Biomarkers , Bone Remodeling/physiology , Periodontal Diseases/physiopathology , Cathepsin K/physiology , Periodontal Diseases/enzymology , Periodontal Diseases/immunology , Alkaline Phosphatase/analysis , Bone Matrix/physiology , Bone Resorption/physiopathology , Saliva/enzymology
18.
J. appl. oral sci ; 24(2): 153-161, Mar.-Apr. 2016. graf
Article in English | LILACS | ID: lil-779903

ABSTRACT

ABSTRACT Objective Biocompatible materials such as interconnected porous hydroxyapatite ceramics (IP-CHA) loaded with osteogenic cells and bioactive agents are part of an evolving concept for overcoming craniofacial defects by use of artificial bone tissue regeneration. Amongst the bioactive agents, melatonin (MEL) and basic fibroblast growth factor (FGF-2) have been independently reported to induce osteoblastic activity. The present in vitro study was undertaken to examine the relationship between these two bioactive agents and their combinatory effects on osteoblastic activity and mineralization in vitro. Material and Methods Mouse preosteoblast cells (MC3T3-E1) were seeded and cultured within cylindrical type of IP-CHA block (ø 4x7 mm) by vacuum-assisted method. The IP-CHA/MC3T3 composites were subjected to FGF-2 and/or MEL. The proliferation assay, alkaline phosphatase enzyme activity (ALP), mRNA expressions of late bone markers, namely Osteocalcin (OCN) and Osteopontin (OPN), and Alizarin Red staining were examined over a period of 7 days. Results FGF-2 mainly enhanced the proliferation of MC3T3-E1 cells within the IP-CHA constructs. MEL mainly induced the mRNA expression of late bone markers (OCN and OPN) and showed increased ALP activity of MC3T3 cells cultured within IP-CHA construct. Moreover, the combination of FGF-2 and MEL showed increased osteogenic activity within the IP-CHA construct in terms of cell proliferation, upregulated expressions of OCN and OPN, increased ALP activity and mineralization with Alizarin Red. The synergy of the proliferative potential of FGF-2 and the differentiation potential of MEL showed increased osteogenic activity in MC3T3-E1 cells cultured within IP-CHA constructs. Conclusion These findings indicate that the combination of FGF-2 and MEL may be utilized with biocompatible materials to attain augmented osteogenic activity and mineralization.


Subject(s)
Animals , Mice , Osteoblasts/drug effects , Fibroblast Growth Factor 2/pharmacology , Durapatite/pharmacology , Bone Substitutes/pharmacology , Melatonin/pharmacology , Time Factors , Materials Testing , Calcification, Physiologic/drug effects , Microscopy, Electron, Scanning , Cell Differentiation/drug effects , Cell Survival/drug effects , Cells, Cultured , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction , Cell Proliferation/drug effects , Alkaline Phosphatase/analysis
19.
J. appl. oral sci ; 24(1): 76-84, Jan.-Feb. 2016. graf
Article in English | LILACS, BBO | ID: lil-777354

ABSTRACT

ABSTRACT Objective The purpose of this study was to investigate the biological effects of epicatechin (ECN), a crosslinking agent, on human dental pulp cells (hDPCs) cultured in collagen scaffolds. Material and Method To evaluate the effects of ECN on the proliferation of hDPCs, cell counting was performed using optical and fluorescent microscopy. Measurements of alkaline phosphatase (ALP) activity, alizarin red staining, and real-time polymerase chain reactions were performed to assess odontogenic differentiation. The compressive strength and setting time of collagen scaffolds containing ECN were measured. Differential scanning calorimetry was performed to analyze the thermal behavior of collagen in the presence of ECN. Results Epicatechin increased ALP activity, mineralized nodule formation, and the mRNA expression of dentin sialophosphoprotein (DSPP), a specific odontogenic-related marker. Furthermore, ECN upregulated the expression of DSPP in hDPCs cultured in collagen scaffolds. Epicatechin activated the extracellular signal-regulated kinase (ERK) and the treatment with an ERK inhibitor (U0126) blocked the expression of DSPP. The compressive strength was increased and the setting time was shortened in a dose-dependent manner. The number of cells cultured in the ECN-treated collagen scaffolds was significantly increased compared to the cells in the untreated control group. Conclusions Our results revealed that ECN promoted the proliferation and differentiation of hDPCs. Furthermore, the differentiation was regulated by the ERK signaling pathway. Changes in mechanical properties are related to cell fate, including proliferation and differentiation. Therefore, our study suggests the ECN treatment might be desirable for dentin-pulp complex regeneration.


Subject(s)
Humans , Catechin/pharmacology , Collagen/pharmacology , Cross-Linking Reagents/pharmacology , Dental Pulp/cytology , Dental Pulp/drug effects , Tissue Scaffolds/chemistry , Time Factors , Calorimetry, Differential Scanning , Gene Expression , Cell Differentiation/drug effects , Cell Survival/drug effects , Cells, Cultured , Blotting, Western , Reproducibility of Results , Analysis of Variance , Extracellular Signal-Regulated MAP Kinases/analysis , Cell Proliferation/drug effects , Alkaline Phosphatase/analysis , Real-Time Polymerase Chain Reaction
20.
J. appl. oral sci ; 23(5): 467-471, Sept.-Oct. 2015. graf
Article in English | LILACS, BBO | ID: lil-764155

ABSTRACT

Mineral Trioxide Aggregate (MTA) is a calcium silicate-based material. New sealers have been developed based on calcium silicate as MTA Fillapex and MTA Plus.Objective The aim of this study was to evaluate biocompatibility and bioactivity of these two calcium silicate-based sealers in culture of human dental pulp cells (hDPCs).Material and Methods The cells were isolated from third molars extracted from a 16-year-old patient. Pulp tissue was sectioned into fragments with approximately 1 mm3 and kept in supplemented medium to obtain hDPCs adherent cultures. Cell characterization assays were performed to prove the osteogenic potential. The evaluated materials were: MTA Plus (MTAP); MTA Fillapex (MTAF) and FillCanal (FC). Biocompatibility was evaluated with MTT and Neutral Red (NR) assays, after hDPCs exposure for 24 h to different dilutions of each sealer extract (1:2, 1:3 and 1:4). Unexposed cells were the positive control (CT). Bioactivity was assessed by alkaline phosphatase (ALP) enzymatic assay in cells exposed for one and three days to sealer extracts (1:4 dilution). All data were analyzed by ANOVA and Tukey post-test (p≤0.05%).Results MTT and NR results showed suitable cell viability rates for MTAP at all dilutions (90-135%). Cells exposed to MTAF and FC (1:2 and 1:4 dilutions) showed significant low viability rate when compared to CT in MTT. The NR results demonstrated cell viability for all materials tested. In MTAP group, the cells ALP activity was similar to CT in one and three days of exposure to the material. MTAF and FC groups demonstrated a decrease in ALP activity when compared to CT at both periods of cell exposure.Conclusions The hDPCs were suitable for the evaluation of new endodontic materialsin vitro. MTAP may be considered a promising material for endodontic treatments.


Subject(s)
Humans , Adolescent , Aluminum Compounds , Biocompatible Materials , Calcium Compounds , Cell Survival/drug effects , Dental Pulp/drug effects , Oxides , Root Canal Filling Materials , Silicates , Alkaline Phosphatase/analysis , Alkaline Phosphatase/drug effects , Analysis of Variance , Barium Sulfate , Bismuth , Borates , Cells, Cultured , Drug Combinations , Eugenol , Formazans , Materials Testing , Reproducibility of Results , Resins, Synthetic , Statistics, Nonparametric , Tetrazolium Salts , Time Factors , Zinc Oxide
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