Application of cutinase in the degradation of biodegradable polyester poly(butylene adipate-co-terephthalate) / 生物工程学报

With the environmental pollution caused by waste plastics becoming increasingly serious, biodegradable polyester has become the focus of public attention. Poly(butylene adipate-co-terephthalate) (PBAT) is a biodegradable polyester formed by the copolymerization of aliphatic and aromatic groups, which has excellent performance of both. The degradation of PBAT under natural conditions requires strict environmental conditions and long degradation cycle. To address these shortcomings, this study explored the application of cutinase in PBAT degradation and the impact of butylene terephthalate (BT) content on the biodegradability of PBAT, so as to improve the degradation rate of PBAT. Five Polyester degrading enzymes from different sources were selected to degrade PBAT to pick out the most efficient enzyme. Subsequently, the degradation rate of PBAT materials with different BT content were determined and compared. The results showed that cutinase ICCG was the best enzyme for PBAT biodegradation, and the higher the BT content, the lower the degradation rate of PBAT. Furthermore, the optimum temperature, buffer type, pH, the ratio of enzyme to substrate (E/S) and substrate concentration in the degradation system were determined to be 75 ℃, Tris HCl, 9.0, 0.4% and 1.0% respectively. These findings may facilitate the application of cutinase in PBAT degradation.

Isolation and identification of a polyester-polyurethane degrading bacterium Bacillus altitudinis YX8-1 / 生物工程学报

Although polyurethane (PUR) plastics play important roles in daily life, its wastes bring serious environmental pollutions. Biological (enzymatic) degradation is considered as an environmentally friendly and low-cost method for PUR waste recycling, in which the efficient PUR-degrading strains or enzymes are crucial. In this work, a polyester PUR-degrading strain YX8-1 was isolated from the surface of PUR waste collected from a landfill. Based on colony morphology and micromorphology observation, phylogenetic analysis of 16S rDNA and gyrA gene, as well as genome sequence comparison, strain YX8-1 was identified as Bacillus altitudinis. The results of high performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) showed that strain YX8-1 was able to depolymerize self-synthesized polyester PUR oligomer (PBA-PU) to produce a monomeric compound 4, 4'-methylene diphenylamine. Furthermore, strain YX8-1 was able to degrade 32% of the commercialized polyester PUR sponges within 30 days. This study thus provides a strain capable of biodegradation of PUR waste, which may facilitate the mining of related degrading enzymes.

Magnesium versus poly-L-lactic acid bioresorbable scaffolds: in vivo optical coherence tomography comparison of mechanical performance / Andamiajes bioresorbibles de magnesio versus poliméricos: comparación in vivo de su comportamiento mecánico por OCT

Abstract Background: Different mechanical properties have been suggested for metallic bioresorbable vascular scaffolds (BVS) in comparison to polymeric BVS. We aim to evaluate the acute mechanical performance of Magmaris® scaffold in comparison to Absorb®. Materials and Methods: Two groups of 10 coronary lesions treated with Magmaris® and Absorb® 1.1 (20584 vs. 21016 struts) were compared. In all cases, optical coherence tomographic (OCT) images were acquired after scaffold deployment. Baseline clinical, angiographic, and procedural characteristics were compared, including OCT evaluations. Results: No baseline clinical or angiographic significant differences were found between groups. The most common indication for revascularization was effort angina (60% vs. 70% p = 0.45) with no ST-elevation myocardial infarction (MI) cases. Main target artery was left anterior descending, with a mean vessel diameter of 3.46 ± 0.23 in Absorb® and 3.52 ± 0.19mm in Magmaris® groups (p = 0.56). All cases underwent pre- and post-dilatation with a procedural success rate of 100%. OCT analyses showed larger scaffold and vessel diameters in Magmaris® group: 3.11 ± 0.38 mm versus 3.07 ± 0.36 mm, p = 0.03 and 4.12 ± 0.51 mm versus 4.04 ± 0.46 mm, p = 0.04. Despite the application of slightly higher postdilatation pressures to Magmaris® devices (18.01 ± 2.15 vs. 17.20 ± 3.80 atm, p = 0.05), significantly lower percentages of disrupted and malapposed struts were identified within Magmaris® scaffolds (0.15% vs. 0.27%, p = 0.03 and 1.06% vs. 1.46% p = 0.01). No cardiac death, target vessel-related MI, or clinically driven target lesion revascularization was reported in a 30-day follow-up. Conclusion: Mechanical properties of Magmaris® scaffold allow achieving larger vessel and scaffold diameters in a safe manner, with lower rates of malapposition and scaffold disruption.

Resumen Introducción: Se ha sugerido la presencia de un distinto comportamiento mecánico entre los dos grupos principales de dispositivos bioresorbibles: metálicos y poliméricos. En este estudio evaluamos el comportamiento mecánico agudo del andamiaje bioresorbible metálico Magmaris® frente al del polimérico Absorb®. Métodos: Se compararon dos grupos de 10 lesiones coronarias tratadas con Magmaris® y Absorb® 1.1 (20584 vs. 21016 struts). En todos los casos se realizó estudio postimplante del dispositivo mediante tomografia de coherencia óptica (OCT). Se compararon las características basales clínicas y angiográficas, así como aspectos del procedimiento (incluídos los estudios de OCT) entre ambos grupos. Resultados: No se encontraron diferencias clínicas o angiográficas estadísticamente significativas entre ambos grupos. La indicación más frecuente de revascularización coronaria fué la presencia de angina de esfuerzo (60% vs. 70% p = 0.45), sin incluirse casos de IAMCEST. La arteria descendente anterior fué el principal vaso diana, con un diámetro medio de 3.46 ± 0.23 mm en el grupo de Absorb® y de 3.52 ± 0.19mm en el grupo de Magmaris® (p = 0.56). En todos los casos se realizó pre y postdilatación, con una tasa de éxito del procedimiento del 100%. Los estudios mediante OCT demostraron un mayor diámetro de stent y del vaso en el grupo de Magmaris®: 3.11 ± 0.38mm versus 3.07 ± 0.36 mm, p = 0.03 y 4.12 ± 0.51mm versus 4.04 ± 0.46mm, p = 0.04. A pesar de someter a los dispositivos Magmaris® a presiones de postdilatación ligeramente superiores (18.01 ± 2.15 vs. 17.20 ± 3.80 atm, p = 0.05), se identificó un menor porcentaje estadísticamente significativo de struts rotos o malapuestos en dicho grupo (0.15% vs. 0.27 %, p = 0.03 y 1.06 % vs. 1.46% p = 0.01). En un seguimiento a 30 días no se registraron eventos mayores: muerte cardíaca, IM relacionado con vaso diana o TLR. Conclusión: Las propiedades mecánicas del scaffold metálico bioresorbible Magmaris® permiten alcanzar mayores diámetros de stent y vaso de forma segura tras su implante, con una baja tasa de malaposición y disrupción.

Statistical analysis of shrinkage levels of human brain slices preserved by sheet plastination technique with polyester resin / Análisis estadístico de los niveles de retracción de cortes de cerebro humano conservados mediante la técnica de plastinación de cortes con resina poliéster

Plastination is currently the most important anatomical preservation technique due to the possibility of preserving bodies and organs for an indefinite period, in a dry and biosecure form, while preserving the morphological characteristics of the tissues. However, the shrinkage of the samples is also part of the plastination, perhaps becoming one of its few disadvantages. This paper presents the shrinkage caused by the classic technique of sheet plastination with polyester resin (Biodur® P40) in human brain slices, with the aim of statistically establishing the percentages of tissue shrinkage caused by this plastination protocol.

La plastinación es actualmente la técnica de preservación anatómica más importante debido a la posibilidad de preservar los cuerpos y órganos por un período indefinido, en forma seca y biosegura, al tiempo que preserva las características morfológicas de los tejidos. Sin embargo, la retracción de las muestras también es parte de la plastinación, quizás convirtiéndose en una de sus pocas desventajas. Este artículo presenta la retracción causada por la técnica clásica de plastinación de cortes con resina poliéster (Biodur® P40) en cortes de cerebro humano, con el objetivo de establecer estadísticamente los porcentajes de retracción de tejidos causados por este protocolo de plastinación.

Desarrollo de un protocolo de plastinación de cortes con resina poliéster aplicado a secciones de cerebro humano / Development of a sheet plastination protocol with polyester resin applied to human brain slices

La plastinación es una técnica anatómica de conservación cadavérica creada en 1977 por Gunther von Hagens, en Heidelberg, Alemania, y que sustituye los líquidos biológicos y/o de fijación por acetona, para luego impregar las muestras con distintas resinas, dependiendo de la técnica de plastinación desarrollada, para finalmente llevar a cabo la polimerización de los componentes incorporados a las muestras, para obtener muestras biológicas secas y totalmente duraderas. El objetivo de este trabajo consistió en desarrollar un protocolo de plastinación de cortes con resina poliéster (Biodur® P40) en secciones de 3 mm de espesor de cerebro humano. La muestras fueron fijadas y conservadas con formalina al 10 %. Los cerebros luego fueron seccionados con una maquina cortadora de tejidos, obteniéndose láminas delgadas de 3 mm de espesor. Inmediatamente los cortes de cerebro fueron colocados en deshidratación en acetona al 100 %, a -25 ºC, durante 7 días el primer baño de acetona, y durante otros 3 días más, para el segundo baño de acetona. Una vez deshidratados los cortes, estos fueron colocados en resina poliéster Biodur® P40 y se llevó a cabo la impregnación forzada de los cortes, en cámara de vacío a temperatura ambiente (20 ºC). Una vez finalizada la impregnación forzada, se procedió a la etapa de curado, la cual en primer lugar consiste en el armado de las cámaras de curado dentro de las cuales se colocaran los cortes con resina poliéster. Las cámaras de curado fueron colocadas bajo luz UV para acelerar la polimerización del poliéster y finalizar el proceso de plastinación. Se logró desarrollar satisfactoriamente en el Laboratorio de Plastinación y Técnicas Anatómicas de la Universidad de La Frontera un protocolo de plastinación de cortes con resina poliéster, obteniendo una excelente conservación de cortes de cerebro, con diferenciación de sustancias gris y blanca, y conservación de todas las características morfológicas.

Plastination is an anatomical technique of cadaveric conservation created in 1977 by Gunther von Hagens, in Heidelberg, Germany, and that substitutes biological and / or fixation fluids with acetone, to then impregnate the samples with different resins, depending on the developed plastination technique, to finally carry out the polymerization of the components incorporated into the samples, to obtain dry and totally durable biological samples. The aim of this work was to develop a sheet plastination protocol with polyester resin (Biodur® P40) in 3 mm thick slices of human brain. The samples were fixed and preserved with 10 % formalin. The brains were sectioned with a slice cut machine, obtaining thin sheets of 3 mm thick. Immediately the slices of brain were placed in dehydration in 100 % acetone, at -25 °C, for 7 days the first acetone bath, and for another 3 more days, for the second acetone bath. Once the cuts were dehydrated, they were placed in Biodur® P40 polyester resin and the forced impregnation was carried out in a vacuum chamber at room temperature (20 °C). Once the forced impregnation was finished, the curing stage was carried out, which first consists in the assembly of the curing chambers within which the slices with polyester resin were placed. The curing chambers were placed under UV light to accelerate the polymerization of the polyester and finished the plastination process. A sheet plastination protocol with polyester resin was successfully developed in the Laboratory of Plastination and Anatomical Techniques of Universidad de La Frontera, obtaining excellent conservation of brain slices, with differentiation of gray and white substances, and conservation of all morphological characteristics.

Proliferation and differentiation of mouse spermatogonial stem cells on a three-dimensional surface composed of PCL/gel nanofibers / Proliferación y diferenciación de células madre espermatogónicas de ratón en una superficie tridimensional compuesta de nanofibras PCL / gel

Spermatogonial stem cells (SSCs) have self-renewal and differentiation capacity essential for sperm production throughout the male reproductive life. The electrospun polycaprolactone/gelatin (PCL/Gel) nanofibrous scaffold mimics important features of the extracellular matrix (ECM), which can provide a promising technique for the proliferation and differentiation of SSCs in vitro. The goal of the present study was to investigate the effects of PCL/Gel nanofibrous scaffold on the propagation and differentiation of neonate mouse SSCs (mSSCs). mSSCs were enzymatically isolated, and the cells were purified by differential plating method and seeded on scaffold. After 2 weeks, viability, colony number and diameter, and expression of specific spermatogonial cell genes were investigated. After mSSCs propagation, the cells were cultivated in a differentiation medium on the scaffold for another 2 weeks, and differentiating cells were analyzed by real-time PCR. The number of mSSC colony (P<0.01) and expression levels of specific spermatogonial genes Plzf and Inga6 (P<0.01) and also differentiation genes c-Kit, Tp1 and Ptm1 (P<0.05) were higher in scaffold group compared with control during the culture period. We conclude that mSSCs can be expanded and can differentiate toward spermatid cells on PCL/Gel nanofibrous scaffold with improved developmental parameters.

Las células madre espermatogónicas (CME) tienen capacidad de auto renovación y diferenciación esenciales para la producción de esperma a lo largo de la vida reproductiva masculina. El «scaffold¼ nanofibroso de policaprolactona / gelatina (PCL / Gel) electrohilado imita características importantes de la matriz extracelular (MEC), que puede proporcionar una técnica prometedora para la proliferación y diferenciación de CME in vitro. El objetivo del presente estudio fue investigar los efectos del «scaffold¼ nanofibroso PCL / Gel en la propagación y diferenciación de CME de ratones neonatos (mSSC). Los mSSC se aislaron enzimáticamente y las células se purificaron mediante un método de siembra diferencial y se sembraron en un «scaffold¼. Después de 2 semanas, se investigaron la viabilidad, el número y el diámetro de las colonias y la expresión de genes específicos de células espermatogónicas. Después de la propagación de mSSC, las células se cultivaron en un medio de diferenciación en el «scaffold¼ durante otras 2 semanas, y las células se analizaron mediante PCR en tiempo real. El número de colonias mSSC (P <0,01) y los niveles de expresión de los genes espermatogónicos específicos Plzf e Inga6 (P <0,01) y también los genes de diferenciación c-Kit, Tp1 y Ptm1 (P <0,05) fueron mayores en el grupo de «scaffold¼ en comparación con el control durante el período de cultivo. Concluimos que los mSSC pueden expandirse y diferenciarse en células espermátidas en un «scaffold¼ de nanofibras PCL / Gel con parámetros de desarrollo mejorados.

Poly(trimethylene carbonate-co-L-lactide) electrospun scaffolds for use as vascular grafts

Currently, there is great clinical need for suitable synthetic grafts that can be used in vascular diseases. Synthetic grafts have been successfully used in medium and large arteries, however, their use in small diameter vessels is limited and presents a high failure rate. In this context, the aim of this study was to develop tissue engineering scaffolds, using poly(trimethylene carbonate-co-L-lactide) (PTMCLLA), for application as small diameter vascular grafts. For this, copolymers with varying trimethylene carbonate/lactide ratios - 20/80, 30/70, and 40/60 - were submitted to electrospinning and the resulting scaffolds were evaluated in terms of their physicochemical and biological properties. The scaffolds produced with PTMCLLA 20/80, 30/70, and 40/60 showed smooth fibers with an average diameter of 771±273, 606±242, and 697±232 nm, respectively. When the degradation ratio was evaluated, the three scaffold groups had a similar molecular weight (Mw) on the final day of analysis. PTMCLLA 30/70 and 40/60 scaffolds exhibited greater flexibility than the PTMCLLA 20/80. However, the PTMCLLA 40/60 scaffolds showed a large wrinkling and their biological properties were not evaluated. The PTMCLLA 30/70 scaffolds supported the adhesion and growth of mesenchymal stem cells (MSCs), endothelial progenitor cells, and smooth muscle cells (SMCs). In addition, they provided a spreading of MSCs and SMCs. Given the results, the electrospun scaffolds produced with PTMCLLA 30/70 copolymer can be considered promising candidates for future applications in vascular tissue engineering.

Síntesis de nanopartículas de ácido poliláctico cargadas con antibióticos y su actividad antibacteriana contra Escherichia coli O157:H7 y Staphylococcus aureus resistente a meticilina / Synthesis of antibiotic loaded polylactic acid nanoparticles and their antibacterial activity against Escherichia coli O157:H7 and methicillin-resistant Staphylococcus aureus

Resumen Introducción: Las nanopartículas poliméricas constituyen una herramienta nanotecnológica que podría ayudar a combatir los microorganismos patógenos que han desarrollado resistencia a los antibióticos convencionales. Objetivo: Sintetizar nanopartículas de ácido poliláctico cargadas con ofloxacina y vancomicina, y determinar su actividad antibacteriana frente a Escherichia coli O157:H7 y Staphylococcus aureus resistente a la meticilina (SARM). Materiales y métodos: Las nanopartículas de ácido poliláctico cargadas con ofloxacina y vancomicina se sintetizaron utilizando el método de emulsión y evaporación de solvente. Se caracterizaron mediante dispersión de luz en modo dinámico, electroforesis Doppler con láser y microscopía electrónica de barrido (S-TEM). Se evaluó la actividad antibacteriana in vitro de las nanopartículas de ácido poliláctico con ofloxacina contra E. coli O157:H7 y nanopartículas de ácido poliláctico con vancomicina contra SARM, mediante el método de microdilución en caldo. Resultados: Se obtuvieron nanopartículas poliméricas con tamaños inferiores a 379 nm y carga superficial positiva de hasta 21 mV. Las nanopartículas cargadas con ofloxacina presentaron una concentración inhibitoria mínima (CIM50) de 0,001 μg/ml frente a E. coli O157:H7, valor 40 veces menor que la concentración de antibiótico libre necesaria para lograr el mismo efecto (CIM50=0,04 μg/ml). Para SARM, las nanopartículas mejoraron la potencia farmacológica in vitro de la vancomicina al exhibir una MIC50 de 0,005 μg/ml, comparada con la de 0,5 μg/ml del antibiótico libre. Conclusiones: Se mejoró el efecto antibacteriano de la ofloxacina y la vancomicina incorporadas en la matriz polimérica de ácido poliláctico. Las nanopartículas poliméricas constituirían una alternativa para el control de cepas bacterianas de interés en salud pública.

Abstract Introduction: Polymeric nanoparticles are promising nanotechnology tools to fight pathogenic bacteria resistant to conventional antibiotics. Objective: To synthesize polylactic acid nanoparticles loaded with ofloxacin and vancomycin, and to determine their antibacterial activity against Escherichia coli O157:H7 and methicillin-resistant Staphylococcus aureus (MRSA). Materials and methods: We synthesized ofloxacin or vancomycin loaded polylactic acid nanoparticles by the emulsification-solvent evaporation method, and characterized them by dynamic light scattering, laser Doppler electrophoresis and scanning electron microscopy. We evaluated in vitro antibacterial activity of ofloxacin- and vancomycin-loaded polylactic acid nanoparticles against E. coli O157:H7 and MRSA using the broth microdilution method. Results: Ofloxacin- and vancomycin-loaded polylactic acid nanoparticles registered a positive surface charge density of 21 mV and an average size lower than 379 nm. In vitro minimum inhibitory concentration (MIC50) of ofloxacin-polylactic acid nanoparticles was 0,001 μg/ml against E. coli O157:H7, i.e., 40 times lower than the free ofloxacin (MIC50: 0.04 μg/ml), indicating enhanced antibacterial activity while the in vitro MIC50 of vancomycin-polylactic acid nanoparticles was 0,005 μg/ml against MRSA, i.e., 100 times lower than that of free vancomycin (MIC50: 0.5 μg/ml). Conclusion: Polylactic acid nanoparticles loaded with ofloxacin and vancomycin showed a higher antibacterial activity. Polymeric nanoparticles are a possible alternative for drug design against pathogenic bacterial strains of public health interest.

Prevalência de saúde cardiovascular ideal na população brasileira - Pesquisa Nacional de Saúde (2013) / Ideal cardiovascular health prevalence in the Brazilian population - National Health Survey (2013)

RESUMO: A prevenção primordial é definida como a prevenção inicial de fatores de risco, por meio da adoção de comportamentos mais saudáveis. Dentro desse conceito, a American Heart Association (AHA) definiu sete métricas, baseadas em evidências, para se alcançar uma saúde cardiovascular (SCV) ideal. O objetivo deste trabalho foi avaliar a prevalência de SCV na população brasileira, segundo sexo, faixa etária e região de moradia, utilizando os dados da última Pesquisa Nacional de Saúde (PNS), de 2013. Foram avaliados, como preconizado pela AHA, de forma conjunta (número de fatores) e isolada, quatro fatores comportamentais (tabagismo, atividade física, índice de massa corporal e dieta) e três biológicos (pressão arterial, glicemia e níveis de colesterol). A população brasileira atingiu prevalências menores de 1%, de sete fatores em nível ideal. Isoladamente, 3,2% da população apresentaram a dieta em nível ideal, seguido da atividade física (23,6%) e índice de massa corporal (43,7%). A população entre 18 e 35 anos apresentou a maior prevalência de número de métricas conjuntas em nível ideal (0,5%), valor também atingido pela população geral da Região Norte. Os resultados indicam que devem ser realizados ainda maiores esforços por meio de políticas públicas de prevenção primordial para atingir metas adequadas de SCV na população brasileira.

ABSTRACT: Primordial prevention is defined as the initial prevention of risk factors, through the adoption of healthier behaviors. Within this concept, the American Heart Association (AHA) has defined seven metrics, based on evidence, to achieve ideal cardiovascular health. The aim of this study was to evaluate the prevalence of cardiovascular health in the Brazilian population, according to sex, age, and region of residence, using data from the latest National Health Survey (2013). We assessed the risk factors, as recommended by the AHA, combined (number of factors) and individually: four behavioral (smoking, physical activity, body mass index and diet) and three biological factors (blood pressure, blood glucose and cholesterol levels). The Brazilian population has reached very low prevalence (1%), for the sum of 7 factors in ideal level. Individually, 3.2% of the population consumed ideal diet, followed by physical activity (23.6%) and body mass index (43.7%). The subjects aged between 18 and 35 years showed higher prevalence of metrics combined at the optimal levels (0.5%), which was also reached by the population of the Northern region. These results indicate that greater efforts are urgent by public policies at the level of primordial prevention in order to achieve appropriate targets of cardiovascular health in the Brazilian population.

Influence of Poly-L-Lactic Acid Scaffold's Pore Size on the Proliferation and Differentiation of Dental Pulp Stem Cells

The aim of this study was to evaluate the influence of the poly-L-lactic acid (PLLA)-based scaffold's pore size on the proliferation and differentiation of dental pulp stem cells (DPSCs). The scaffolds were prepared in pulp chambers of 1-mm-thick tooth slices from third molars using salt crystals (150-250 µm or 251-450 µm) as porogen. DPSC (1x105 cells) were seeded in the scaffolds with different pore sizes, and cultured in 24-well plates. The cell proliferation was evaluated using the WST-1 assay after 3-21 days. Furthermore, RT-PCR was used to assess the differentiation of the DPSCs into odontoblasts, using markers of odontoblastic differentiation (DSPP, DSP-1 and MEPE). RNA from human odontoblasts was used as control. Cell proliferation rate was similar in both scaffolds except at the 14th day period, in which the cells seeded in the scaffolds with larger pores showed higher proliferation (p<0.05). After 21 days DPSCs seeded in both evaluated scaffolds were able of expressing odontoblastic markers DMP-1, DSPP and MEPE. In summary, both scaffolds tested in this study allowed the proliferation and differentiation of DPSCs into odontoblast-like cells.

O objetivo desse estudo foi avaliar a influência do tamanho dos poros de um scaffold à base de poli ácido láctico (PLLA) sobre a proliferação e diferenciação de células tronco da polpa dental (dental pulp stem cells - DPSC). Os scaffolds foram preparados dentro da câmara pulpar de discos de terceiros molares (1 mm), utilizando sal como porógeno (150-250 µm ou 251-450 µm). DPSC (1x105 células) foram semeadas nos scaffolds com diferentes tamanhos de poros e cultivadas em placas de 24 poços. A proliferação celular foi avaliada utilizando WST-1 após 3-21 dias. Além disso, RT-PCR foi utilizado para avaliar a diferenciação odontoblástica das DPSC utilizando marcadores da diferenciação odontoblástica (DSPP, DMP-1 e MEPE). RNA obtido de odontoblastos humanos foi utilizado como controle. A taxa de proliferação celular foi semelhante nos dois scaffolds avaliados, exceto no 14° dia, no qual as células cultivadas nos scaffolds com os maiores poros apresentaram uma maior taxa de proliferação (p<0,05). Após 21 dias, as DSPC cultivadas em ambos scaffolds avaliados foram capazes de expressar os marcadores odontoblásticos DMP-1, DSPP e MEPE. Em resumo, ambos scaffolds avaliados nesse estudo permitiram a proliferação e diferenciação odontoblástica das DPSC. .

Penetration of a resin-based filling material into lateral root canals and quality of obturation by different techniques

The aim of this study was to evaluate the penetration of a resin/polyester polymer-based material (Resilon Real Seal; SybronEndo Corp., Orange, USA) into simulated lateral canals, and the quality of obturations by different techniques. A total of 30 standardized simulated canals were divided into three groups according to the technique of obturation used: MS (McSpadden), SB (SystemB/Obtura II), and LC (Lateral Condensation). To analyze the penetration of the filling material, the simulated canals were digitalized and the images were analyzed using the Leica QWIN Pro v2.3 software. The data of the middle and apical thirds were separately submitted to analysis of variance (ANOVA), followed by the Tukey’s test for the comparison of the techniques. Results showed a significant difference (p < 0.05) between groups (LC < SB) in the middle third, and a significant difference (p < 0.05) between groups (LC < SB and MS < SB) in the apical third. To analyze the quality of the obturations, the canals were radiographed and evaluated by three examiners. The Kappa test on interexaminer agreement and the nonparametric Kruskal-Wallis test indicated no significant difference between filling techniques. It was concluded that Resilon achieves greater levels of penetration when associated with thermoplastic obturation techniques.

Poly (l-lactide acid) improves complete nano-hydroxyapatite bone scaffolds through the microstructure rearrangement

Cracks often occur when nano-hydroxyapatite bone scaffolds are fabricated with selective laser sintering, which affect the performance of scaffolds. In this study, a small amount of poly (l-lactide acid) (PLLA) was added into nano-hydroxyapatite (nano-HAP) powder by mechanical blending in order to improve the sintering properties. The nano-HAP powder combined with 1wt percent PLLA was sintered under different laser power (5W, 7.5W, 10W, 12.5W, 15W and 20W). The fabricated scaffolds were characterized using Scanning Electron Microscope (SEM), X-ray Diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR) and Micro Hardness Tester. The results showed that nano-HAP particles grew up quickly with the laser power increasing, and there were many strip-like cracks on the surface of sintering zone. The cracks gradually reduced until disappeared when the laser power increased to 15W, together with a great improvement of density. Large pores were observed on the specimen when the laser power further increases, accompanied with the decomposition of HAP into β-tricalcium phosphate (β-TCP) and tetracalcium phosphate (TTCP). And the optimum parameters were eventually obtained with laser power of 15W, scanning speed of 1000 mm/min, powder bed temperature of 150ºC, laser spot diameter of 2 mm and layer thickness of 0.2 mm. We summarized that the molten PLLA enhanced the particle rearrangement of nano-HAP by capillary force and may absorb thermal stress in laser sintering process, while PLLA would be oxidized gradually until completely excluded from the sintered nano-HAP scaffolds, which was confirmed by FTIR analysis. This study provides a novel method to improve the sintering properties of nano-HAP with no adverse effects which would be used in the application of bone tissue engineering potentially.

In vitro comparative study of white and dark polycaprolactone trifumarate in situ cross-linkable scaffolds seeded with rat bone marrow stromal cells

OBJECTIVE: Dark poly(caprolactone) trifumarate is a successful candidate for use as a bone tissue engineering scaffold. Recently, a white polymeric scaffold was developed that shows a shorter synthesis time and is more convenient for tissue-staining work. This is an in vitro comparative study of both the white and dark scaffolds. METHODS: Both white and dark poly(caprolactone) trifumarate macromers were characterized via Fourier transform infrared spectroscopy before being chemically cross-linked and molded into disc-shaped scaffolds. Biodegradability was assessed by percentage weight loss on days 7, 14, 28, 42 and 56 (n = 5) after immersion in 10% serum-supplemented medium or distilled water. Static cell seeding was employed in which isolated and characterized rat bone marrow stromal cells were seeded directly onto the scaffold surface. Seeded scaffolds were subjected to a series of biochemical assays and scanning electron microscopy at specified time intervals for up to 28 days of incubation. RESULTS: The degradation of the white scaffold was significantly lower compared with the dark scaffold but was within the acceptable time range for bone-healing processes. The deoxyribonucleic acid and collagen contents increased up to day 28 with no significant difference between the two scaffolds, but the glycosaminoglycan content was slightly higher in the white scaffold throughout 14 days of incubation. Scanning electron microscopy at days 1 and 14 revealed cellular growth and attachment. CONCLUSIONS: There was no cell growth advantage between the two forms, but the white scaffold had a slower biodegradability rate, suggesting that the newly synthesized poly(caprolactone) trifumarate is more suitable for use as a bone tissue engineering scaffold.

Effects of combined mechanical stimulation on the proliferation and differentiation of pre-osteoblasts

We observed how combined mechanical stimuli affect the proliferation and differentiation of pre-osteoblasts. For this research, a bioreactor system was developed that can simultaneously stimulate cells with cyclic strain and ultrasound, each of which is known to effectively stimulate bone tissue regeneration. MC3T3-E1 pre-osteoblasts were chosen for bone tissue engineering due to their osteoblast-like characteristics. 3-D scaffolds were fabricated with polycaprolactone and poly-L-lactic acid using the salt leaching method. The cells were stimulated by the bioreactor with cyclic strain and ultrasound. The bioreactor was set at a frequency of 1.0 Hz and 10% strain for cyclic strain and 1.0 MHz and 30 mW/cm2 for ultrasound. Three experimental groups (ultrasound, cyclic strain, and combined stimulation) and a control group were examined. Each group was stimulated for 20 min/day. Mechanical stimuli did not affect MC3T3-E1 cell proliferation significantly up to 10 days when measured with the cell counting kit-8. However, gene expression analysis of collagen type-I, osteocalcin, RUNX2, and osterix revealed that the combined mechanical stimulation accelerated the matrix maturation of MC3T3-E1 cells. These results indicate that the combined mechanical stimulation can enhance the differentiation of pre-osteoblasts more efficiently than simple stimuli, in spite of no effect on cell proliferation.

Isolation and identification of poly beta hydroxybutyric acid accumulating bacteria of Staphylococcal sp. and characterization of biodegradable polyester.

Staphylococcus sp. strain BP/SU1, capable of degrading the biopolymer and utilize it as a source of carbon and energy, was isolated from activated sludge using METABOLIX (MBX D411G). It was found that this strain was capable of accumulating poly(3-hydroxybutyric acid) P(3-HB), as granule poly (3-hydroxybutyric acid), p(3-HB), inclusion bodies when grown under suitable nutrient conditions. These strains could sustain cell growth up to a dry mass of 9.24 g/l with a doubling time of 8 to 10 hr and could accumulate P(3-HB) as granular inclusion bodies to a cell dry weight of more than 12%. P(3-HB) accumulated by this organism was isolated and characterized through NMR, FT-IR spectroscopy, UV Spectroscopy, Mass spectroscopy and Differential Scanning Calorimetry. P(3-HB) granules so isolated showed physical and chemical properties that should be possessed by a superior quality thermoplastic biopolymer.

Análisis de las técnicas de impresión en prótesis parcial removible a extensión distal / Analysis of the impression technique in distal extension removable partial denture

El procedimiento de impresión es una etapa importante en la elaboración de la prótesis parcial removible al reproducir la anatomía dentaria, los rebordes alveolares residuales y las estructuras adyacentes. La impresión funcional se indica para los casos dentomucosoportados para hacer compatible las diferencias en el comportamiento de los dientes pilares y la mucosa del reborde bajo cargas funcionales. Existen distintas técnicas de impresiones funcionales y todas ellas influyen distintamente en el desplazamiento de los tejidos que recubren al reborde y en el comportamiento de las bases protésicas. La técnica de modelo alterado es uno de los procedimientos de impresión más utilizados debido a los resultados que se obtienen, sin embargo, recientemente se ha demostrado efectos similares con otras técnicas.

The impression procedure is an important step in achieving sucess in removable partial denture. The functional impression is used for tooth-mucosa supported removable partial denture and it´s an attempt to accommodate the difference in resiliency between hard and soft tissue support. There are many techniques and all of them displace residual ridge mucosa in different ways. The altered cast impression fulfills the requirements of functional impression, however, other techniques have demonstrated the same effects.

Surface roughness of composite resins after finishing and polishing

This study evaluated the effect of surface finishing methods on the average surface roughness of resin composites. Seven composites and two polishing systems were used. One hundred and twenty-six conical specimens of each material were prepared in stainless steel molds against a polyester strip. Forty-two of them remained intact and were used as controls. Each half of the remaining samples was polished with either diamond burs or diamond burs + aluminum oxide discs. The results showed no statistical difference in average surface roughness (Ra, µm) between the polyester strip and aluminum oxide discs (p>0.05). However, finishing with diamond burs showed a statistically higher average roughness for all composites (p<0.05). Statistical differences were detected among materials (p<0.05) in the use of diamond burs

Comparative study of tissue response to polyglecaprone 25, polyglactin 910 and polytetrafluorethylene suture materials in rats

The authors evaluated the biocompatibility of three suture materials, polyglecaprone 25, polyglactin 910 and polytetrafluorethylene, implanted into subcutaneous tissue in the dorsal region of 20 Wistar albinus rats. After periods of 2, 7, 14 and 21 days, the rats were sacrificed and the specimens were processed for routine histotechnical analysis and stained with hematoxylin and eosin. The rate of fibrosis, angioblastic and fibroblastic proliferation, and also the intensity of inflammation were observed with the optic microscope. The results showed that polyglecaprone 25 suture material induced a mild inflammatory reaction, followed by polyglactin 910 and polytetrafluorethylene, respectively. Such biological behavior must be considered during the selection of the suture material to be used in oral surgery.

Molecular weight dependence of intrinsic viscosity for polyarylates at 30°C

The intrinsic viscosity of polyarylates having different molecular weights was calculated in O-chlorophenol at 30C. The root-mean-square distance [h 2] 1/2 separating the ends of polyarylates chain was calculated. Analysis of data indicated that the intrinsic viscosity is proportional to the average molecular weight and [h 2] 1/2. The slopes of Stock-Mayer and Fixman plots were used to determine the mutual excluded volume [beta] of segments. The beta-values obtained were compared with the values calculated for the poly[nitroarylates] and were found to be in the order: Beta poly[nitroarylates] > > beta[polyarylates]. This was due to the presence of the nitro group and the ether linkage in the backbone of the polyarylates

Spontaneous ignition behaviour of cotton fabric having different amounts of polyester

Measurements were made on a series of cottonpolyester blends having different compositions. The results showed that the time to ignition increases as the polyester contents of the fabric were increased and explanation of this observation is given. Both the orientation of the fabric and the tewperature of the furnace environment have affected the time to ignition values. The time to ignition was found to be longer for the fabric loaded in horizontal position than that loaded vertical position. This difference was attributed to the effect of the forced convection induced inside the furnace