Scanning electron microscopic analysis of the effect of Carisolv TM gel on periodontally compromised human root surfaces

The aim of this study was to analyze, under scanning electron microscopy (SEM), the morphologic characteristics of root surfaces after application of CarisolvTM gel in association with scaling and root planing (SRP). Sixty periodontally compromised extracted human teeth were randomly assigned to 6 groups: 1) SRP alone; 2) passive topical application of CarisolvTM + SRP; 3) active topical application of CarisolvTM + SRP; 4) multiple applications of CarisolvTM + SRP; 5) SRP + 24% EDTA; 6) topical application of CarisolvTM + SRP + 24% EDTA. CarisolvTM gel was applied to root surfaces for 30 s, followed by scaling and root planing, consisting of 50 strokes with Gracey curettes in an apical-coronal direction, parallel to the long axis of the tooth. The only exception was group 4, in which the roots were instrumented until a smooth, hard and glass-like surface was achieved. All specimens were further analyzed by SEM. The results showed that the treatment with CarisolvTM caused significant changes in root surface morphology of periodontally compromised teeth only when the chemical agent was actively applied (burnishing technique). CarisolvTM failed to remove the smear layer completely, especially with a single application, independently of the method of application. Multiple applications of CarisolvTM were necessary to achieve a smear layer reduction comparable to that obtained with 24% EDTA conditioning.

Scanning electron microscopic and profilometric study of different sharpening stones

Scaling and root planing contribute to the recovery of periodontal health. All periodontal instruments loose their fine cutting angle after use. To maintain this angle, correct sharpening is required using specifically designed stones. The characteristics of sharpening stones can be compared to the blade of the instruments and also transported to root surface during instrumentation. Root smoothness is related to the quality of the blade. Therefore, the purpose of this study was to evaluate the characteristics of 9 sharpening stones by scanning electron microscopic and profilometric analyses. Ceramic and Neumar stones were very fine and both may be recommended to maintain the sharpness of the instruments. Arkansas, Thompson and CE stones presented greater roughness with very regular and round particles, and are suitable for maintenance of the cutting angle. In addition, these stones may be indicated for the routine sharpening of the instruments that are partly dull. Oxide Aluminum, Carborundum and JON stones were the coarsest with large irregular particles and may be indicated for initial sharpening of totally dull instruments with completion of sharpening with finer stones.

Scanning electron microscope assessment of several resharpening techniques on the cutting edges of gracey curettes

Aim: Treatment of periodontal diseases is based on efficient scaling and root planing (SRP) and adequate maintenance of the patient. The effectiveness of SRP is influenced by operator skill, access to the subgingival area, root anatomy, and the quality and type of instrument used for SRP. The aim of this study was to evaluate the cutting edges of Gracey curettes after manufacturing and after resharpening using several techniques. Methods and Material: The cutting edges of a total of 41 new #5-6 stainless steel Gracey curettes were evaluated blindly using scanning electron microscopy (SEM). The quality of the cutting edges was evaluated blindly by a calibrated examiner using micrographs. Data were analyzed using a Kruskal Wallis test and nonparametric two-way multiple comparisons. Results and Conclusions: Different sharpening techniques had significantly different effects on the sharpeness of cutting edges (p<0.05). Sharpening by passing the lateral face of curettes over a sharpening stone and then a #299 Arkansas stone produced a high frequency of smooth, sharp edges or slightly irregular edges between the lateral and coronal faces of the curettes. Sharpening by passing a blunt stone over the curette's lateral face produced the poorest quality cutting edge (a bevel). Sharpening of the coronal curette face produced extremely irregular cutting edges and non-functional wire edges. Sharpening with rotary devices produced extremely irregular cutting edges.

A scanning electronic microscopy study of the action of different desensitizing agents on bovine dentin

This study sought to use scanning electronic microscopy (SEM) to evaluate the dentinal tubule occlusion potential of different desensitizing agents. Ten slices of bovine dentin were divided into six fragments, cleaned (using ultrasound), and etched for 15 seconds with a 35% phosphoric acid solution. All but one of the groups received a different desensitizing agent; the sixth group served as a control and received no additional treatment. After the agents were applied, the dentin specimens were analyzed by SEM and scores were assigned based on the extent of tubular obliteration. Only three agents demonstrated tubular sealing that was significantly different from that of the control group.

Scanning electron microscopic study of the in situ effect of salivary stimulation on erosion and abrasion in human and bovine enamel

This in situ study investigated, using scanning electron microscopy, the effect of stimulated saliva on the enamel surface of bovine and human substrates submitted to erosion followed by brushing abrasion immediately or after one hour. During 2 experimental 7-day crossover phases, 9 previously selected volunteers wore intraoral palatal devices, with 12 enamel specimens (6 human and 6 bovine). In the first phase, the volunteers immersed the device for 5 minutes in 150 ml of a cola drink, 4 times a day (8h00, 12h00, 16h00 and 20h00). Immediately after the immersions, no treatment was performed in 4 specimens (ERO), 4 other specimens were immediately brushed (0 min) using a fluoride dentifrice and the device was replaced into the mouth. After 60 min, the other 4 specimens were brushed. In the second phase, the procedures were repeated but, after the immersions, the volunteers stimulated the salivary flow rate by chewing a sugar-free gum for 30 min. Enamel superficial alterations of all specimens were then evaluated using a scanning electron microscope. Enamel prism core dissolution was seen on the surfaces submitted to erosion, while on those submitted to erosion and to abrasion (both at 0 and 60 min) a more homogeneous enamel surface was observed, probably due to the removal of the altered superficial prism layer. For all the other variables - enamel substrate and salivary stimulation the microscopic pattern of the enamel specimens was similar.

Scanning electron microscopic evaluation of the root apex of mandibular premolars.

This aim of this study was to evaluate the root apex of mandibular premolars regarding the presence of main and accessory foramina. The root apexes from fifty extracted mandibular single-rooted premolars were examined by scanning electron microscopy (SEM). The apical openings had their diameter measured and were identified as main or accessory foramina. Double blinded and calibrated examiners analyzed the SEM photographs and classified the premolar roots into three types, based on the presence and size of the apical openings. Type I: roots with a single main apical foramen and no accessory foramina; type II: roots with a main foramen and one or more accessory foramina; type III: roots with accessory foramina only. For the first premolar, 16 roots were classified as type I (48.48%), 4 as type II (12.12%) and 13 as type III (39.40%). For the second premolars, 10 roots were classified as type I (58.83%), 3 as type II (17.65%) and 4 as type III (23.52%). The high incidence of roots with accessory foramina only (type III), mainly in the first premolar, warns of the need for caution during working length determination and apical debridement.

Simultaneous two-wavelength transmission quantitative phase microscopy with a color camera.

We present a quantitative phase microscopy method that uses a Bayer mosaic color camera to simultaneously acquire off-axis interferograms in transmission mode at two distinct wavelengths. Wrapped phase information is processed using a two-wavelength algorithm to extend the range of the optical path delay measurements that can be detected using a single temporal acquisition. We experimentally demonstrate this technique by acquiring the phase profiles of optically clear microstructures without 2pi ambiguities. In addition, the phase noise contribution arising from spectral channel crosstalk on the color camera is quantified.

A porcine deep dermal partial thickness burn model with hypertrophic scarring

We developed a reproducible model of deep dermal partial thickness burn injury in juvenile Large White pigs. The contact burn is created using water at 92 degrees C for 15s in a bottle with the bottom replaced with plastic wrap. The depth of injury was determined by a histopathologist who examined tissue sections 2 and 6 days after injury in a blinded manner. Upon creation, the circular wound area developed white eschar and a hyperaemic zone around the wound border. Animals were kept for 6 weeks or 99 days to examine the wound healing process. The wounds took between 3 and 5 weeks for complete re-epithelialisation. Most wounds developed contracted, purple, hypertrophic scars. On measurement, the thickness of the burned skin was approximately 1.8 times that of the control skin at week 6 and approximately 2.2 times thicker than control skin at 99 days after injury. We have developed various methods to assess healing wounds, including digital photographic analysis, depth of organising granulation tissue, immunohistochemistry, electron microscopy and tensiometry. Immunohistochemistry and electron microscopy showed that our porcine hypertrophic scar appears similar to human hypertrophic scarring. The development of this model allows us to test and compare different treatments on burn wounds.

Synthesis of biodegradable polymer-mesoporous silica composite microspheres for DNA prime-protein boost vaccination

DNA vaccines or proteins are capable of inducing specific immunity; however, the translation to the clinic has generally been problematic, primarily due to the reduced magnitude of immune response and poor pharmacokinetics. Herein we demonstrate a composite microsphere formulation, composed of mesoporous silica spheres (MPS) and poly(d,l-lactide-co-glycolide) (PLGA), enables the controlled delivery of a prime-boost vaccine via the encapsulation of plasmid DNA (pDNA) and protein in different compartments. Method with modified dual-concentric-feeding needles attached to a 40 kHz ultrasonic atomizer was studied. These needles focus the flow of two different solutions, which passed through the ultrasonic atomizer. The process synthesis parameters, which are important to the scale-up of composite microspheres, were also studied. These parameters include polymer concentration, feed flowrate, and volumetric ratio of polymer and pDNA-PEI/MPS-BSA. This fabrication technique produced composite microspheres with mean D[4,3] ranging from 6 to 34 μm, depending upon the microsphere preparation. The resultant physical morphology of composite microspheres was largely influenced by the volumetric ratio of pDNA-PEI/MPS-BSA to polymer, and this was due to the precipitation of MPS at the surface of the microspheres. The encapsulation efficiencies were predominantly in the range of 93-98% for pDNA and 46-68% for MPS. In the in vitro studies, the pDNA and protein showed different release kinetics in a 40 day time frame. The dual-concentric-feeding in ultrasonic atomization was shown to have excellent reproducibility. It was concluded that this fabrication technique is an effective method to prepare formulations containing a heterologous prime-boost vaccine in a single delivery system.

Large-volume methacrylate monolith for plasmid purification : process engineering approach to synthesis and application

The extent of exothermicity associated with the construction of large-volume methacrylate monolithic columns has somewhat obstructed the realisation of large-scale rapid biomolecule purification especially for plasmid-based products which have proven to herald future trends in biotechnology. A novel synthesis technique via a heat expulsion mechanism was employed to prepare a 40 mL methacrylate monolith with a homogeneous radial pore structure along its thickness. Radial temperature gradient was recorded to be only 1.8 °C. Maximum radial temperature recorded at the centre of the monolith was 62.3 °C, which was only 2.3 °C higher than the actual polymerisation temperature. Pore characterisation of the monolithic polymer showed unimodal pore size distributions at different radial positions with an identical modal pore size of 400 nm. Chromatographic characterisation of the polymer after functionalisation with amino groups displayed a persistent dynamic binding capacity of 15.5 mg of plasmid DNA/mL. The maximum pressure drop recorded was only 0.12 MPa at a flow rate of 10 mL/min. The polymer demonstrated rapid separation ability by fractionating Escherichia coli DH5α-pUC19 clarified lysate in only 3 min after loading. The plasmid sample collected after the fast purification process was tested to be a homogeneous supercoiled plasmid with DNA electrophoresis and restriction analysis.

Mesoporous silica spheres from colloids

A novel method has been developed to synthesize mesoporous silica spheres using commercial silica colloids (SNOWTEX) as precursors and electrolytes (ammonium nitrate and sodium chloride) as destabilizers. Crosslinked polyacrylamide hydrogel was used as a temporary barrier to obtain dispersible spherical mesoporous silica particles. The influences of synthesis conditions including solution composition and calcination temperature on the formation of the mesoporous silica particles were systematically investigated. The structure and morphology of the mesoporous silica particles were characterized via scanning electron microscopy (SEM) and N2 sorption technique. Mesoporous silica particles with particle diameters ranging from 0.5 to 1.6 μm were produced whilst the BET surface area was in the range of 31-123 m2 g-1. Their pore size could be adjusted from 14.1 to 28.8 nm by increasing the starting particle diameter from 20-30 nm up to 70-100 nm. A simple and cost effective method is reported that should open up new opportunities for the synthesis of scalable host materials with controllable structures.

The suitability of DEAE-Cl active groups on customized poly(GMA-co-EDMA) continuous stationary phase for fast enzyme-free isolation of plasmid DNA

The creation of a commercially viable and a large-scale purification process for plasmid DNA (pDNA) production requires a whole-systems continuous or semi-continuous purification strategy employing optimised stationary adsorption phase(s) without the use of expensive and toxic chemicals, avian/bovine-derived enzymes and several built-in unit processes, thus affecting overall plasmid recovery, processing time and economics. Continuous stationary phases are known to offer fast separation due to their large pore diameter making large molecule pDNA easily accessible with limited mass transfer resistance even at high flow rates. A monolithic stationary sorbent was synthesised via free radical liquid porogenic polymerisation of ethylene glycol dimethacrylate (EDMA) and glycidyl methacrylate (GMA) with surface and pore characteristics tailored specifically for plasmid binding, retention and elution. The polymer was functionalised with an amine active group for anion-exchange purification of pDNA from cleared lysate obtained from E. coli DH5α-pUC19 pellets in RNase/protease-free process. Characterization of the resin showed a unique porous material with 70% of the pores sizes above 300 nm. The final product isolated from anion-exchange purification in only 5 min was pure and homogenous supercoiled pDNA with no gDNA, RNA and protein contamination as confirmed with DNA electrophoresis, restriction analysis and SDS page. The resin showed a maximum binding capacity of 15.2 mg/mL and this capacity persisted after several applications of the resin. This technique is cGMP compatible and commercially viable for rapid isolation of pDNA.

Avaliação da customização por desgaste de pinos pré-fabricados de fibra de vidro

O objetivo deste estudo foi avaliar pinos pré-fabricados de fibra de vidro (White Post DC/FGM) submetidos à customização por desgaste da porção apical. Experimento 1: 5 pinos n. 4 foram divididos em 5 grupos (G) de acordo com o instrumento de desgaste: GA - sem desgaste, GB- mini torno industrial (Dentsply), GC - ponta diamantada n. 3195F (KG Sorensen), GD - disco de lixa de granulação média (Sof-Lex/3M/ESPE), GE- alicate (Tramontina). Observou-se a micromorfologia dos pinos em microscópio eletrônico de varredura (ZEISS/DSM 960). Experimento 2: 60 pinos de diferentes diâmetros foram divididos em 6 grupos: G0 - pinos n. 0,5, G1 - pinos n. 1, G2 - pinos n. 2, G3 - pinos n. 3, G4 - pinos n.4, G5 - pinos n. 4 com terço apical desgastado com discos de lixa até o equivalente ao terço apical dos pinos n. 2. Os pinos foram submetidos ao teste de flexão de 3 pontos na máquina de ensaios universal (Instron 5500 R), conforme ISO 10477. Experimento 3: 20 caninos humanos permanentes sofreram tratamento endodôntico e remoção das coroas clínicas padronizando 15 mm de remanescente radicular. Os dentes foram incluídos em resina acrílica com simulação do ligamento periodontal, receberam férula de 2 mm e foram divididos em 2 grupos: GI - pinos n. 4 cimentados em condutos preparados com broca equivalente ao pino (FGM), GII - pinos n. 4 customizados no terço apical cimentados em condutos preparados com brocas (FGM) equivalentes aos pinos n. 2 em 10 mm e n. 4 em 5 mm. Os pinos foram cimentados com cimento resinoso (Rely X U100/3M/ESPE), os corpos de prova receberam coroas diretas de resina composta (Enforce Core/Dentsply) padronizadas com coras de policarbonato (TDV) e foram submetidos ao teste de resistência à fratura na Instron a 45da ferramenta cilíndrica, com força de 500 N aplicada a 2 mm da incisal na face palatina/lingual, com velocidade de 0,5 mm/min até falha. O padrão de fratura foi classificado em favorável ou desfavorável. Os resultados foram tratados estatisticamente por teste de análise de variância (ANOVA, p<0,05). Os resultados dos testes de flexão e fratura foram respectivamente: G0 - 58,406,40; G1 - 83,959,43; G2- 103,4219,17; G3 - 160,7817,30; G4 - 170,4711,28; G5 - 106,3521,96; GI - 303,0262,21 e GII - 402,81131,97. O padrão de fratura foi tratado por Mann-Whitney que observou semelhança estatística entre os grupos. Concluiu-se que o desgaste de pinos de fibra de vidro com pontas diamantadas ou discos de lixa produz alterações micromorfológicas aceitáveis. O corte com alicate deve ser evitado. A customização por desgaste da porção apical de pinos de fibra de vidro diminui a resistência à flexão a valores aceitáveis. Dentes restaurados com pinos de fibra de vidro customizados por desgaste possuem resistência à fratura superior a dentes restaurados com pinos intactos. A customização por desgaste facilita a adaptação do pino ao conduto radicular e preserva a estrutura dental.

Functional properties of cell-seeded three-dimensionally woven poly(epsilon-caprolactone) scaffolds for cartilage tissue engineering.

Articular cartilage possesses complex mechanical properties that provide healthy joints the ability to bear repeated loads and maintain smooth articulating surfaces over an entire lifetime. In this study, we utilized a fiber-reinforced composite scaffold designed to mimic the anisotropic, nonlinear, and viscoelastic biomechanical characteristics of native cartilage as the basis for developing functional tissue-engineered constructs. Three-dimensionally woven poly(epsilon-caprolactone) (PCL) scaffolds were encapsulated with a fibrin hydrogel, seeded with human adipose-derived stem cells, and cultured for 28 days in chondrogenic culture conditions. Biomechanical testing showed that PCL-based constructs exhibited baseline compressive and shear properties similar to those of native cartilage and maintained these properties throughout the culture period, while supporting the synthesis of a collagen-rich extracellular matrix. Further, constructs displayed an equilibrium coefficient of friction similar to that of native articular cartilage (mu(eq) approximately 0.1-0.3) over the prescribed culture period. Our findings show that three-dimensionally woven PCL-fibrin composite scaffolds can be produced with cartilage-like mechanical properties, and that these engineered properties can be maintained in culture while seeded stem cells regenerate a new, functional tissue construct.

Inkless microcontact printing on SAMs of Boc- and TBS-protected thiols.

We report a new inkless catalytic muCP technique that achieves accurate, fast, and complete pattern reproduction on SAMs of Boc- and TBS-protected thiols immobilized on gold using a polyurethane-acrylate stamp functionalized with covalently bound sulfonic acids. Pattern transfer is complete at room temperature just after one minute of contact and renders sub-200 nm size structures of chemically differentiated SAMs.