Effect of root canal preparation, type of endodontic post and mechanical cycling on root fracture strength

Objective: To evaluate the impact of the type of root canal preparation, intraradicular post and mechanical cycling on the fracture strength of roots. Material and Methods: Eighty human single rooted teeth were divided into 8 groups according to the instruments used for root canal preparation (manual or rotary instruments), the type of intraradicular post (fiber posts-FRC and cast post and core-CPC) and the use of mechanical cycling (MC) as follows: Manual and FRC; Manual, FRC and MC; Manual and CPC; Manual, CPC and MC; Rotary and FRC; Rotary, FRC and MC; Rotary and CPC; Rotary, CPC and MC. The filling was performed by lateral compactation. All root canals were prepared for a post with a 10 mm length, using the custom # 2 bur of the glass fiber post system. For mechanical cycling, the protocol was applied as follows: an angle of incidence of 45 degrees, 37 degrees C, 88 N, 4 Hz, 2 million pulses. All groups were submitted to fracture strength test in a 45 degrees device with 1 mm/min cross-head speed until failure occurred. Results: The 3-way ANOVA showed that the root canal preparation strategy (p<0.03) and post type (p<0.0001) affected the fracture strength results, while mechanical cycling (p=0.29) did not. Conclusion: The root canal preparation strategy only influenced the root fracture strength when restoring with a fiber post and mechanical cycling, so it does not seem to be an important factor in this scenario

The effect of cholesterol on the reconstitution of alkaline phosphatase into liposomes

Tissue-nonspecific alkaline phosphatase (TNAP), present on the surface of chondrocyte- and osteoblast-derived matrix vesicles (MVs), plays key enzymatic functions during endochondral ossification. Many studies have shown that MVs are enriched in TNAP and also in cholesterol compared to the plasma membrane. Here we have studied the influence of cholesterol on the reconstitution of TNAP into dipalmitoylphosphatidylcholine (DPPC)-liposomes, monitoring the changes in lipid critical transition temperature (T(c)) and enthalpy variation (Delta H) using differential scanning calorimetry (DSC). DPPC-liposomes revealed a T(c) of 41.5 degrees C and Delta H of 7.63 Kcal mol(-1). The gradual increase in cholesterol concentration decrease Delta H values, reaching a Delta H of 0.87 Kcal mol(-1) for DPPC: cholesterol system with 36 mol% of cholesterol. An increase in T(c), up to 47 degrees C for the DPPC:cholesterol liposomes (36 mol% of Chol), resulted from the increase in the area per molecule in the gel phase. TNAP (0.02 mg/mL) reconstitution was done with protein:lipid 1:10,000 (molar ratio), resulting in 85% of the added enzyme being incorporated. The presence of cholesterol reduced the incorporation of TNAP to 42% of the added enzyme when a lipid composition of 36 mol% of Chol was used. Furthermore, the presence of TNAP in proteoliposomes resulted in a reduction in Delta H. The gradual proportional increase of cholesterol in liposomes results in broadening of the phase transition peak and eventually eliminates the cooperative gel-to-liquid-crystalline phase transition of phospholipids bilayers. Thus, the formation of microdomains may facilitate the clustering of enzymes and transporters known to be functional in MVs during endochondral ossification. (C) 2010 Elsevier B.V. All rights reserved.FAPESPCAPESCNP

NPP1 and TNAP hydrolyze ATP synergistically during biomineralization

Matrix vesicles (MVs) are a special class of extracellular vesicles released by mineralizing cells during bone and tooth mineralization that initiate the precipitation of apatitic minerals by regulating the extracellular ratio between inorganic phosphate (P-i), a calcification promoter, and pyrophosphate (PPi), a calcification inhibitor. The P-i/PPi ratio is thought to be controlled by two ecto-phosphatases present on the outer leaflet of the MVs' membrane: ectonucleotide pyrophosphatase/ phosphodiesterase 1 (NPP1) that produces PPi as well as P-i from ATP and tissue-nonspecific alkaline phosphatase (TNAP) that hydrolyzes both ATP and PPi to generate P-i. However, if and how these enzymes act in concert in MVs are still unclear. Herein, we investigated the role of NPP1 and TNAP in ATP hydrolysis during MV-mediated biomineralization using proteoliposomes as a biomimetic model for MVs. Proteoliposomes composed by 1,2-dipalmitoylphosphatidylcholine (DPPC) and harboring NPP1 alone, TNAP alone, or both together at different molar ratios (1:1, 10:1, and 1:10) were fabricated. After 48 h of incubation with ATP, TNAP-containing proteoliposomes consumed more ATP than NPP1-containing vesicles (270 and 210 nmol, respectively). Both types of vesicles comparatively formed ADP (205 and 201 nmol, respectively), while NPP1-containing vesicles hydrolyzed AMP less efficiently than TNAP-containing proteoliposomes (10 and 25 nmol, respectively). In vitro mineralization assays showed that in the presence of ATP, TNAP-harboring proteoliposomes mineralized through a sigmoidal single-step process, while NPP1-harboring vesicles displayed a two-step mineralization process. ATR-FTIR analyses showed that the minerals produced by TNAP-harboring proteoliposomes were structurally more similar to hydroxyapatite than those produced by NPP1-harboring vesicles. Our results with proteoliposomes indicate that the pyrophosphohydrolase function of NPP1 and the phosphohydrolase activity of TNAP act synergistically to produce a P-i/PPi ratio conducive to mineralization and the synergism is maximal when the two enzymes are present at equimolar concentrations. The significance of these findings for hypophosphatasia is discussed

Canal Preparation and Filling Techniques do not Influence the Fracture Resistance of Extensively Damaged Teeth

The purpose of this study was to assess the fracture resistance of extensively damaged teeth after two root canal preparation techniques (hand and rotary files) and after two filling techniques (active and passive compaction). Sixty-eight maxillary canines roots with an apical diameter equal to that of a #25 K-file were embedded in acrylic resin and the periodontal ligament was simulated by using a polyether impression material. The roots were randomly distributed into four groups (n=17): hand preparation and active compaction (HA), hand preparation and passive compaction (HP), rotary preparation and active compaction (RA), and rotary preparation and passive compaction (RP). All roots were restored with glass fiber post and metallic crown. The specimens were mechanically cycled (500,000 cycles, 45°, 37°C, 133 N, 2 Hz) and then subjected to a fracture resistance test. A single blinded examiner analyzed the external root surface and classified the failure pattern as favorable or unfavorable. The fracture resistance values ranged between 621.15 N (HP) and 785.71 N (HA). However, the Kruskal-Wallis test did not reveal differences in the fracture resistance values among the four groups (p =0.247). Under the tested conditions, root canal preparation and filling techniques had no influence on the fracture resistance of extensively damaged teeth restored with fiber post and metallic crown.O objetivo deste estudo foi avaliar a resistência à fratura de dentes amplamente destruídos após duas técnicas de preparo (limas manual e rotatória) e após duas técnicas obturadoras (compactação ativa e passiva). Sessenta e oito raízes de caninos superiores com diâmetro apical igual a uma lima K #25 foram embutidas em resina acrílica e o ligamento periodontal foi simulado utilizando um material de moldagem à base de poliéter. As raízes foram distribuídas aleatoriamente em quatro grupos (n=17): preparo manual e compactação ativa (MA), preparo manual e compactação passiva (MP), preparo rotatório e compactação ativa (RA) e preparo rotatório e compactação passiva (RP). Todas as raízes foram restauradas com pino de fibra de vidro e coroa metálica. Os espécimes foram ciclados mecanicamente (500.000 ciclos, 45°, 37°C, 133 N, 2 Hz) e depois submetidos ao teste de resistência à fratura. Um único examinador cego analisou a superfície externa das raízes e classificou o padrão de falha em favorável ou desfavorável. Os valores de resistência à fratura variaram entre 621,15 N (MP) e 785,71 N (MA). Entretanto, o teste de Kruskal-Wallis não revelou diferença nos valores de resistência à fratura entre os quatro grupos (p=0,247). Diante das condições testadas, as técnicas de preparo e de obturação do canal radicular não influenciam na resistência à fratura de dentes amplamente destruídos restaurados com pino de fibra de vidro e coroa metálica