Effect of genetic polymorphisms rs2301113 and rs2057482 in the expression of HIF-1α protein in periodontal ligament fibroblasts subjected to compressive force

Abstract Objective Many genes and signaling molecules are involved in orthodontic tooth movement, with mechanically and hypoxically stabilized HIF-1α having been shown to play a decisive role in periodontal ligament signaling during orthodontic tooth movement. Thus, this in vitro study aimed to investigate if genetic polymorphisms in HIF1A (Hypoxia-inducible factor α-subunits) influence the expression pattern of HIF-1α protein during simulated orthodontic compressive pressure. Methodology Samples from human periodontal ligament fibroblasts were used and their DNA was genotyped using real time Polymerase chain reaction for the genetic polymorphisms rs2301113 and rs2057482 in HIF1A . For cell culture and protein expression experiments, six human periodontal ligament fibroblast cell lines were selected based on the patients' genotype. To simulate orthodontic compressive pressure in fibroblasts, a 2 g/cm2 force was applied under cell culture conditions for 48 hours. Protein expression was evaluated by Western Blot. Paired t-tests were used to compare HIF-1α expression with and without compressive pressure application and unpaired t-tests were used to compare expression between the genotypes in rs2057482 and rs2301113 (p<0.05). Results The expression of HIF-1α protein was significantly enhanced by compressive pressure application regardless of the genotype (p<0.0001). The genotypes in the genetic polymorphisms rs2301113 and rs2057482 were not associated with HIF-1α protein expression (p>0.05). Conclusions Our study confirms that compressive pressure application enhances HIF-1α protein expression. We could not prove that the genetic polymorphisms in HIF1A affect HIF-1α protein expression by periodontal ligament fibroblasts during simulated orthodontic compressive force.

Fit of metal-ceramic crowns: effect of coping fabrication method and veneering ceramic application

Aim: To evaluate the influence of cobalt-chromium (Co-Cr) coping fabrication methods and ceramic application on the marginal and internal fit of metal-ceramic crowns. Methods: Co-Cr copings for metal-ceramic crowns were prepared by lost wax casting or CAD-CAM machining of sintered blocks. The fit was analyzed using the silicone replica technique at four assessment points: marginal gap (MG), axial wall (AW), axio-occlusal (AO) angle, and central occlusal (CO) wall. After the initial analysis, the copings were ceramic-veneered with the layering technique, and the fit was again determined. Data were statistically analyzed by paired and unpaired Student's-t test (α=0.05). Results: Marginal and internal fit before ceramic application according to the coping manufacturing method showed significant differences only at CO (p < 0.001), with milled copings (137.98±16.71 µm) showing higher gap values than cast copings (112.86±8.57 µm). For cast copings, there were significant differences at MG (before 109.13±8.79 µm; after 102.78±7.18 µm) and CO (before 112.86±8.57 µm; after 104.07±10.63 µm) when comparing the fit before and after ceramic firing. For milled copings, there was significant difference only at AO (before 116.39±9.64 µm; after 108.54±9.26 µm). Conclusion: This study demonstrated that the coping fabrication method influenced the internal fit. Ceramic firing maintained or improved the fit of the metal-ceramic crowns. The marginal discrepancy of all restorations, before and after ceramic firing, can be considered clinically acceptable

Influence of pre-curing different adhesives on the color stability of cemented thin ceramic veneers

Abstract The objective was to evaluate the color stability of thin ceramic veneers as a function of the curing mode (with and without pre-cure) of different adhesive systems applied to the internal surface of ceramics. Five adhesive systems (Scotchbond Multi-Purpose, Single Bond Universal, Gluma 2 Bond, Ambar, and Ambar APS) and photo-cured resin cement (Allcem Veneer) were used for cementing 0.6 mm-thick feldspathic veneers (Mark II) on composite resin substrates (Charisma Diamond). The groups, according to adhesive system, were divided into two subgroups (n=10): i) adhesive and resin cement were polymerized separately (pre-cure of the adhesive), ii) adhesive and resin cement were polymerized simultaneously. The CIELab color parameters were determined with a spectrophotometer at 24h (baseline), 7 days, 30 days and 12 months. Data were analyzed by two-way ANOVA and Tukey's test (α=0.05). The results indicated statistically significant differences for adhesive systems and time. The mode of curing of the adhesive system was not statistically significant: pre-cured adhesives (2.6±1.3) and not pre-cured adhesives (2.8±1.4). For the adhesives, ΔE values varied in the following order: Ambar-APS (1.6±0.5) < Scotchbond Multi-Purpose (2.6±1.2) = Gluma 2 Bond (2.7±1.2) = Ambar (2.9±1.2) < Single Bond Universal (3.5±1.5). For time, ΔE values were: 7 days (1.7±0.7), 30 days (3.5±1.2) and 12 months (2.9±1.3). It can be concluded that the different adhesive systems used for cementing thin ceramic veneers influenced the final color of the indirect restorations. The adhesives curing mode did not present a significant effect in the color stability of thin ceramic veneers.

Resumo O objetivo deste estudo foi avaliar a estabilidade de cor de laminados cerâmicos de espessura fina em função do momento de ativação de diferentes sistemas adesivos na superfície interna das cerâmicas. Foram utilizados 5 sistemas adesivos (Scotchbond Multiuso, Single Bond Universal, Gluma 2 Bond, Ambar and Ambar APS) e o cimento resinoso fotoativado (Allcem Veneer) para cimentação de lâminas de cerâmica feldspática (Mark II) com 0,6 mm de espessura, sobre substratos de resina composta (Charisma Diamond). Os grupos, de acordo com o sistema adesivo utilizado, foram divididos em dois subgrupos (n=10): i) adesivo e cimento resinoso foram polimerizados separadamente (polimerização isolada do adesivo), ii) adesivo e cimento resinoso foram polimerizados ao mesmo tempo (polimerização simultânea). Os parâmetros de cor do sistema CIELab foram determinados com um espectrofotômetro nos tempos de 24 h (baseline), 7 dias, 30 dias e 12 meses. Os dados foram analisados por ANOVA a dois fatores e teste de Tukey (α=0,05). Os resultados indicaram diferenças estatísticas significantes para os sistemas adesivos e tempo. O momento da fotoativação do sistema adesivo não foi estatisticamente significante: polimerização prévia (2,6±1,3) e polimerização simultânea (2,8±1,4). Para o sistema adesivo, os valores médios de ΔE variaram na seguinte ordem: Ambar APS (1,6±0,5)<Scotchbond Multi-Purpose (2,6±1,2)=Gluma 2 Bond (2,7±1,2)=Ambar (2,9±1,2)<Single Bond Universal (3,5±1,5). Para o fator tempo o ΔE foi: 7 dias (1,7±0,7), 30 dias (3,5±1,2) e 12 meses (2,9±1,3). Pode-se concluir que os diferentes sistemas adesivos utilizados na cimentação de laminados cerâmicos de espessura fina influenciaram na cor final das restaurações. O momento da fotoativação, isolada ou simultânea ao cimento resinoso, não apresentou efeito significativo na estabilidade de cor dos laminados cerâmicos de espessura fina.