RESUMEN
AIM: To compare the bioactivity of Biodentine (BIO, Septodont), MTA Plus (MTA P, Avalon) and calcium silicate experimental cement (CSC) with resin (CSCR) associated with zirconium (CSCR ZrO2 ) or niobium (CSCR Nb2 O5 ) oxide as radiopacifiers. METHODOLOGY: According to the relevance of osteoblastic cell response for mineralized tissue repair, human osteoblastic cells (Saos-2) were exposed to test materials and assessed for viability (MTT), cell proliferation, gene expression of alkaline phosphatase (ALP) osteogenic marker by real-time PCR (RT-qPCR), ALP activity assay and alizarin red staining (ARS) to detect mineralization nodule deposition in osteogenic medium. Unexposed cells acted as the control group (C). Statistical analysis was carried out using ANOVA and the Bonferroni post-test (P < 0.05). RESULTS: All tested cements showed dose-dependent responses in cell viability (MTT). Exposed cells revealed good viability (80-130% compared to the control group) in the highest dilutions of all types of cement. MTA P, BIO and CSCR ZrO2 significantly increased the velocity of cell proliferation after three days of cell exposure in the wound-healing assay (P < 0.05), which corroborated MTT data. On day 3, the ALP transcript level increased, especially to CSCR Nb2 O5 (P < 0.05). All cements exhibited suitable ALP enzyme activity, highlighting the 7-day period of cell exposure. ARS, CSCR Nb2 O5 , revealed a significant potential to induce mineralization in vitro. CONCLUSIONS: All materials had suitable biocompatibility and bioactivity. The MTA P, BIO and CSCR ZrO2 groups had the highest viability rates and velocity of proliferation whilst the CSCR Nb2 O5 group produced more mineralized nodules.
Asunto(s)
Compuestos de Aluminio/farmacología , Materiales Biocompatibles/farmacología , Compuestos de Calcio/farmacología , Cementos Dentales/farmacología , Osteoblastos/efectos de los fármacos , Óxidos/farmacología , Materiales de Obturación del Conducto Radicular/farmacología , Silicatos/farmacología , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Combinación de Medicamentos , Humanos , Ensayo de Materiales , Niobio/farmacología , Circonio/farmacologíaRESUMEN
AIM: To investigate the cytotoxicity, osteogenic bioactivity and mRNA expression of osteogenic markers of bone morphogenetic protein 2 (BMP-2), osteocalcin (OC) and alkaline phosphatase (ALP) induced by the extracts of set MTA Plus (MTA P) (Avalon Biomed Inc. Bradenton, FL, USA) in comparison with MTA (Angelus, Londrina, PR, Brazil) on human dental pulp cells (hDPCs). METHODOLOGY: Cell viability was assessed by mitochondrial dehydrogenase enzymatic (MTT) assay, and the mechanism of cell death was evaluated by flow cytometry. Bioactivity was evaluated by alkaline phosphatase (ALP) assay and detection of calcium deposits with alizarin red staining (ARS). The gene expression of BMP-2, OC and ALP was quantified with real-time PCR. Statistical analysis was performed with analysis of variance and Bonferroni or Tukey post-test (α = 0.05). RESULTS: MTA and MTA P were not cytotoxic and did not induce apoptosis. MTA P had significant higher ALP activity in relation to MTA and the control (P < 0.05). MTA had a significantly higher percentage of mineralized area than MTA P (P < 0.05). The expression of BMP2 and OC mRNA was significantly higher in cells exposed to MTA than MTA P after 1 day (P < 0.05). At day 3, the mRNA expression of ALP was significantly higher in MTA P compared with MTA (P < 0.05). CONCLUSIONS: MTA and MTA Plus were noncytotoxic, increased mineralization processes in vitro and induced the expression of osteogenic markers.
Asunto(s)
Compuestos de Aluminio/farmacología , Compuestos de Calcio/farmacología , Cementos Dentales/farmacología , Pulpa Dental/citología , Osteogénesis/efectos de los fármacos , Osteogénesis/genética , Óxidos/farmacología , Materiales de Obturación del Conducto Radicular/farmacología , Silicatos/farmacología , Adolescente , Adulto , Fosfatasa Alcalina/genética , Apoptosis/efectos de los fármacos , Proteína Morfogenética Ósea 2/genética , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Pulpa Dental/efectos de los fármacos , Combinación de Medicamentos , Expresión Génica/efectos de los fármacos , Humanos , Osteocalcina/genética , ARN Mensajero/genética , Adulto JovenRESUMEN
Although the metabolism of early bovine embryos has not been fully elucidated, several publications have addressed this important issue to improve culture conditions for cattle reproductive biotechnologies, with the ultimate goal of producing in vitro embryos similar in quality to those developing in vivo. Here, we review general aspects of bovine embryo metabolism in vitro and in vivo, and discuss the use of metabolic analysis of embryos produced in vitro to assess viability and predict a viable pregnancy after transference to the female tract.
Asunto(s)
Femenino , Humanos , Masculino , Eficiencia Organizacional/estadística & datos numéricos , Personal de Salud/estadística & datos numéricos , Deficiencia de Vitamina D/sangre , Deficiencia de Vitamina D/epidemiologíaRESUMEN
Although the metabolism of early bovine embryos has not been fully elucidated, several publications have addressed this important issue to improve culture conditions for cattle reproductive biotechnologies, with the ultimate goal of producing in vitro embryos similar in quality to those developing in vivo. Here, we review general aspects of bovine embryo metabolism in vitro and in vivo, and discuss the use of metabolic analysis of embryos produced in vitro to assess viability and predict a viable pregnancy after transference to the female tract.
Asunto(s)
Blastocisto/metabolismo , Metabolismo Energético/fisiología , Fertilización In Vitro , Oocitos/metabolismo , Animales , Dióxido de Carbono/metabolismo , Bovinos , Técnicas de Cultivo de Embriones , Femenino , Glucosa/metabolismo , Ácido Láctico/metabolismo , Consumo de Oxígeno , Embarazo , Ácido Pirúvico/metabolismoRESUMEN
Follicle cultures reproduce in vitro the functional features observed in vivo. In a search for an ideal model, we cultured bovine antral follicle wall sections (FWS) in a serum-free defined medium (DM) known to induce 17ß-estradiol (E2) production, and in a nondefined medium (NDM) containing serum. Follicles were sectioned and cultured in NDM or DM for 24 or 48 h. Morphological features were determined by light microscopy. Gene expression of steroidogenic enzymes and follicle-stimulating hormone (FSH) receptor were determined by RT-PCR; progesterone (P4) and E2 concentrations in the media were measured by radioimmunoassay. DM, but not NDM, maintained an FWS morphology in vitro that was similar to fresh tissue. DM also induced an increase in the expression of all steroidogenic enzymes, except FSH receptor, but NDM did not. In both DM and NDM, there was a gradual increase in P4 throughout the culture period; however, P4 concentration was significantly higher in NDM. In both media, E2 concentration was increased at 24 h, followed by a decrease at 48 h. The E2:P4 ratio was higher in DM than in NDM. These results suggest that DM maintains morphological structure, upregulates the expression of steroidogenic enzyme genes, and maintains steroid production with a high E2:P4 ratio in FWS cultures.
Asunto(s)
Medios de Cultivo/farmacología , Estradiol/farmacología , Folículo Ovárico/efectos de los fármacos , Progesterona/farmacología , Técnicas de Cultivo de Tejidos , Análisis de Varianza , Animales , Aromatasa/genética , Bovinos , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/genética , Medio de Cultivo Libre de Suero , Femenino , Expresión Génica , Folículo Ovárico/anatomía & histología , Fosfoproteínas/genética , Progesterona Reductasa/genética , Receptores de HFE/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Esteroide 17-alfa-Hidroxilasa/genéticaRESUMEN
Follicle cultures reproduce in vitro the functional features observed in vivo. In a search for an ideal model, we cultured bovine antral follicle wall sections (FWS) in a serum-free defined medium (DM) known to induce 17β-estradiol (E2) production, and in a nondefined medium (NDM) containing serum. Follicles were sectioned and cultured in NDM or DM for 24 or 48 h. Morphological features were determined by light microscopy. Gene expression of steroidogenic enzymes and follicle-stimulating hormone (FSH) receptor were determined by RT-PCR; progesterone (P4) and E2 concentrations in the media were measured by radioimmunoassay. DM, but not NDM, maintained an FWS morphology in vitro that was similar to fresh tissue. DM also induced an increase in the expression of all steroidogenic enzymes, except FSH receptor, but NDM did not. In both DM and NDM, there was a gradual increase in P4 throughout the culture period; however, P4 concentration was significantly higher in NDM. In both media, E2 concentration was increased at 24 h, followed by a decrease at 48 h. The E2:P4 ratio was higher in DM than in NDM. These results suggest that DM maintains morphological structure, upregulates the expression of steroidogenic enzyme genes, and maintains steroid production with a high E2:P4 ratio in FWS cultures.