Induction of early neural precursors and derivation of tripotent neural stem cells from human pluripotent stem cells under xeno-free conditions.

Human embryonic stem cells (hESC) and induced pluripotent stem cells (hiPSC) can differentiate into many cell types and are important for regenerative medicine; however, further work is needed to reliably differentiate hESC and hiPSC into neural-restricted multipotent derivatives or specialized cell types under conditions that are free from animal products. Toward this goal, we tested the transition of hESC and hiPSC lines onto xeno-free (XF) / feeder-free conditions and evaluated XF substrate preference, pluripotency, and karyotype. Critically, XF transitioned H9 hESC, Shef4 hESC, and iPS6-9 retained pluripotency (Oct-4 and NANOG), proliferation (MKI67 and PCNA), and normal karyotype. Subsequently, XF transitioned hESC and hiPSC were induced with epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) to generate neuralized spheres containing primitive neural precursors, which could differentiate into astrocytes and neurons, but not oligoprogenitors. Further neuralization of spheres via LIF supplementation and attachment selection on CELLstart substrate generated adherent human neural stem cells (hNSC) with normal karyotype and high proliferation potential under XF conditions. Interestingly, adherent hNSC derived from H9, Shef4, and iPS6-9 differentiated into significant numbers of O4+ oligoprogenitors (∼20-30%) with robust proliferation; however, very few GalC+ cells were observed (∼2-4%), indicative of early oligodendrocytic lineage commitment. Overall, these data demonstrate the transition of multiple hESC and hiPSC lines onto XF substrate and media conditions, and a reproducible neuralization method that generated neural derivatives with multipotent cell fate potential and normal karyotype.

In vitro long-term degradation of aesthetic restorative materials in food-simulating media.

OBJECTIVE: The aim of this study was to evaluate the effect of long-term food-simulating media storage on degradation of restorative materials through roughness measurements. MATERIALS AND METHODS: Sixty cylindrical specimens of each material (Filtek Z250, Esthet X, Filtek Flow, Dyract AP and Vitremer) were prepared, stored for 24 h, and polished. The surface roughness analysis was conducted using Surfcorder (SE1700) roughness-measuring instrument. Three traces were recorded on each specimen at three different locations. The specimens were randomly distributed into five groups (n = 12) according to the storage media: (water, ethanol, Coca-Cola®, citric acid and lactic acid). Roughness measurements were recorded after 1 week, 1, 3 and 6 months. The storage solutions were weekly changed. Data were submitted to ANOVA and Tukey test (p < 0.05). RESULTS: There was no significant increase on roughness means for Filtek Z250, Filtek Flow and Vitremer over time, regardless the storage media. Significant increase on surface roughness was observed for Esthet X after 1 month of storage in Coca-Cola® and after 3 months of storage in citric acid and for Dyract AP after 6 months in water. There were no significant differences on surface roughness between resin composites and compomer. Resin-modified glass ionomer showed a significantly higher surface roughness mean than the other materials. CONCLUSIONS: Food-simulating media affects the surface roughness of the materials. Biodegradation is material-, solution- and time-dependent.

Degradation of polymeric restorative materials subjected to a high caries challenge.

OBJECTIVES: The aim of this study was to evaluate the degradation of different resin filling materials after a caries challenge, by high performance liquid chromatography (HPLC) and contact angle (θ) measurement. METHODS: Four different polymeric restorative materials (a resin composite, a polyacid-modified resin composite, an ormocer and a resin-modified glass ionomer cement) were tested. Five samples (30 mm × 6 mm × 2 mm) of each material were formed in a Teflon mold, following the manufacturer's instructions. After pH cycles, the solutions were injected in an HPLC. The θ was obtained, before and after pH cycle, by a goniometer at 60% air humidity and 25°C. A distilled water drop (0.006 ml) was put on the material surface, and after 6 min, 10 measures were obtained at 20s intervals. Each sample received 4 drops, one at a time, on different areas. RESULTS: HPLC results showed elution of byproducts in all materials. This was greater in the acid medium. Bis-GMA and TEGDMA were detected in TPH Spectrum and Definite residues. Analyses of the contact angle by ANOVA and Student-Neuman-Keuls's test showed that the surfaces of TPH Spectrum, Dyract AP and Definite were altered, except Vitremer (p<0.05). SIGNIFICANCE: All materials tested degraded on a caries simulated medium, suggesting that a great effort should be made to disseminate oral health information, since a high caries challenge environment (low pH) can lead to dental composite degradation, with potential toxic risks to patients.

Aluminium and fluoride release into artificial saliva from dental restoratives placed in teeth.

This study examined the release of aluminium and fluoride from restorative materials placed in either deciduous or young permanent immature teeth stored in artificial saliva for 1 month. Cavities were prepared in extracted teeth, then filled with a fluoride releasing restorative (glass-ionomer, compomer or composite resin), with and without conditioning as appropriate. The teeth were then stored in artificial saliva for 1 month, after which the amount of aluminium and fluoride released was determined spectrophotometrically. With all materials tested, both aluminium and fluoride were released in all cases. Young immature teeth were associated with lower level of ion release which was attributed to the absorption of ions by the enamel. However, unconditioned samples were usually associated with similar ion release to conditioned ones, suggesting that the loss of mineral phase on conditioning has only a marginal effect on the capacity for ion uptake. The ratio of aluminium to fluoride released varied with the type of tooth, deciduous conditioned teeth generally absorbing proportionately less aluminium than young immature teeth. The overall conclusion is that interaction with ions released by restorative materials is influenced by type of tooth.

Efeito do jato profilático de bicarbonato de sódio sobre a superfície de materiais restauradores estéticos / The effect of the prophylactic jet on the surface of restorative materials

Foi avaliado, em Microscopia Eletrônica de Varredura, o efeito do jato profilático de bicarbonato de sódio sobre os materiais: Durafil, Z100, TPH, Concise, Heliomolar Chelon-fil e Dyract AP, com e sem a proteção do selante de superfície Fortify, por 15, 30 ou 60 segundos. Os autores concluíram que a aplicação do jato altera a superfície dos materiais. A partir de 30 segundos, todos os materiais tiveram o contorno alterado com perda de substância

Marginal gap formation of composites in dentine: effect of water storage.

This study investigated the effects of water storage on the marginal adaptation of two composite resins (Spectrum TPH and Ariston AT), two polyacid-modified composite resins (Dyract AP and F2000) and a new PRG composite (Reactmer) to dentine over time. Two cylindrical dentine cavities (1.5 mm diameter and 1.5 mm deep) were prepared on the horizontally sectioned surfaces of freshly extracted teeth. In each tooth, one cavity was restored using composites with their respective bonding system and the other without the bonding system. The sample size for each material with/without bonding system was 6. The composites were placed in one increment, bulk-polymerized, immediately finished/polished with the Sof-lex discs system and fine polished on 30 microm silicon carbide/9 microm aluminium oxide lapping film discs in a microgrinding system. The specimens were then stored in distilled water at 37 degrees C and the maximum marginal gap width between the material and the dentine wall was determined at 24 h, 1 week, 2 weeks, 3 weeks and 4 weeks using a measurescope at x500 magnification. Results were subjected to statistical analysis using monva, anova/Scheffe's post hoc test and independent samples of t-test at significance level 0.05. At all time intervals, no significant difference in marginal gap formation was observed between materials. Where bonding systems were not used, a decrease in gap widths over time was observed with most materials but only Dyract AP exhibited a significant decrease. The latter was observed after 2 weeks storage in water. The use of bonding systems reduced dentine marginal gaps significantly and is therefore mandatory for all composites evaluated. Marginal gaps arising from polymerization contraction of conventional, polyacid-modified and PRG composites cannot be fully compensated by hydroscopic expansion. Clinical relevance Bonding systems are mandatory for all composites when bonding to dentin. In the event of a bond failure at placement, hygroscopic expansion of composites can reduce marginal gaps but not completely.

Avaliaçäo in vitro da liberaçäo de flúor e atividade antimicrobiana de compômeros restauradores / In vitro fluoride release and antimicrobial activity of restorative compomers

Utilizando-se de testes in vitro, os autores avaliaram a liberaçäo de flúor em água deionizada por 14 dias e a atividade antimicrobiana contra o S.mutans # GS-5, de dois compômeros restauradores, o Dyract e o Compoglass. A atividade contra o S.mutans foi medida através do teste de fermentaçäo em caldo por um período de 7 dias, com leitura do pH a cada 24 horas; da quantificaçäo de carboidratos e proteínas depositadas sobre os espécimes; e do teste de difusäo em ágar. O Dyract liberou de 0,039ñ0,011 µgF-/mm² no primeiro dia a 0,019ñ0,004 µgF-/mm², com diferença significante entre ambos a partir do 4º. dia (p<0,05). Os compômeros näo evitaram a fermentaçäo pelo microrganismo, permanecendo o pH abaixo de 4,5 em todos os dias, nem inibiram o crescimento no teste de difusäo. Apenas no teste de produçäo de placa sobre os compômeros e pastilhas de aço, foi verificada menor quantidade de proteina no depósito produzido sobre o Dyract, que no controle (p<0,05). Por esses resultados, os materiais em questäo podem näo ser os mais indicados em situaçöes de risco à cárie