Effect of different obturation techniques on sealer penetration into dentinal tubules after final irrigation with XP-Endo finisher file

The first objective of this research was to evaluate the effectiveness of XP-Endo Finisher on dentinal tubule penetration of irrigation solution using confocal laser scanning microscopy. The main purpose of this research was to compare the effect of cold lateral condensation, continuous wave obturation and core-carrier based techniques on sealer penetration. Sixty mandibular premolars were prepared and allocated into two experimental groups (n=30) as the final irrigation technique and obturation technique experiment. In the final irrigation technique experiment, final irrigation was performed with XP-Endo Finisher, passive ultrasonic irrigation (PUI) and conventional needle irrigation (CNI) (n=10). The roots in the obturation technique experiment were also assigned into 3 groups and obturated with cold lateral condensation, continuous-wave obturation and core-carrier techniques (n=10). The most effective activation method, which emerged as a result of the first part of this study, was used as the final irrigation method in the obturation technique experiment. Then, all roots were sectioned in 1-mm-thick slices at 3mm from the apex for scanning. In terms of depth and percentage of material penetration, CNI exhibited significantly the lowest values and no significant difference was found between others. Also, there was no significant difference among obturation methods. In conclusion, XP-Endo Finisher and PUI are more effective than CNI on irrigant penetration. Sealer penetration into dentinal tubules is independent of obturation techniques.

El objetivo principal de esta investigación fue evaluar la eficacia de XP- Endo Finisher en la penetración de la solución de irrigación en los túbulos dentinarios mediante microscopía de láser confocal. Se prepararon sesenta premolares mandibulares y se distribuyeron en dos grupos experimentales (n=30) según el tipo de método de evaluación utilizado. En el experimento de la técnica de irrigación final, la irrigación final se realizó con XP-Endo Finisher, irrigación ultrasónica pasiva (PUI) e irrigación con aguja convencional (CNI) (n=10). Las raíces en el experimento de la técnica de obturación también se asignaron en 3 grupos y se obturaron con técnicas de condensación lateral fría, obturación de onda continua y portador de núcleo (n=10). El método de activación más eficaz, que surgió como resultado de la primera parte de este estudio, se utilizó como método de irrigación final en el experimento de la técnica de obturación. Luego, todas las raíces se seccionaron en muestras de 1mm de espesor. En términos de profundidad y porcentaje de penetración del material, CNI exhibió significativamente los valores más bajos y no se encontraron diferencias significativas entre los demás. Además, no hubo diferencias significativas entre los métodos de obturación. En conclusión, XP-Endo Finisher y PUI son más efectivos que CNI en la penetración del irrigante. La penetración del sellador en los túbulos dentinarios es independiente de las técnicas de obturación.

Research progress on the methods of observation and analysis of dental bonding interfaces / 口腔疾病防治

@#Dental bonding technology and materials have been used widely in dentistry because of their excellent properties. The development of novel bonding technology and materials is constantly being performed to improve the effect of dental bonding restorations. Observation and analysis of the dental bonding interface is one of the most important methods for laboratory evaluation of bonding efficiency. This paper aims to review the methods of observation and analysis of dental bonding interfaces to provide a reference for the selection of evaluation methods in dental bonding research. The features of 6 methods, including scanning electron microscopy (SEM), transmission electron microscopy (TEM), confocal laser scanning microscopy (CLSM), Raman spectroscopy (RS), optical coherence tomography (OCT) and atomic force microscopy (AFM), were described and summarized. Among these methods, SEM and TEM are used most often in the analysis of fine structures; CLSM and OCT are used for the acquisition of characteristic image signals, such as microleakage and exogenous and endogenous fluorescence; and RS and AFM can test chemical composition and mechanical properties.

Cancer-specific calcium nanoregulator suppressing the generation and circulation of circulating tumor cell clusters for enhanced anti-metastasis combinational chemotherapy

Tumor metastasis is responsible for chemotherapeutic failure and cancer-related death. Moreover, circulating tumor cell (CTC) clusters play a pivotal role in tumor metastasis. Herein, we develop cancer-specific calcium nanoregulators to suppress the generation and circulation of CTC clusters by cancer membrane-coated digoxin (DIG) and doxorubicin (DOX) co-encapsulated PLGA nanoparticles (CPDDs). CPDDs could precisely target the homologous primary tumor cells and CTC clusters in blood and lymphatic circulation. Intriguingly, CPDDs induce the accumulation of intracellular Ca

Dentinal tubule penetration of a calcium silicate-based root canal sealer using a specific calcium fluorophore

Abstract This study aimed to evaluate penetrability on dentinal tubule of a new bioceramic sealer through confocal laser scanning microscopy (CLSM). A specific fluorophore (Fluo-3) was mixed with the sealer. Forty distobuccal roots from maxillary molars were selected, and root canal preparation was carried out with Wave One Gold # 35.06 instruments. Roots were randomly assigned to 4 groups according to the filling procedures: Bioceramic/Lentulo (Sealer Plus BC); Bioceramic/EasyClean group, three activation of the sealer (3x20 s) with Easy Clean instrument; Bioceramic/Irrisonic: ultrasonic activation for 30 s; and AHplus/Lentulo: epoxy resin based sealer (AH Plus) was utilized with the same protocol as the BC/LE group. After 72 h, specimens were transversally sectioned at 2 and 7 mm from root apex and then analyzed through CLSM. Sealer penetration area on dentinal tubule was measured by Adobe Photoshop CC2018. Kruskal Wallis and Wilcoxon T tests were carried out. Penetrability results were similar for both sealers regardless of which technique was performed to activate them inside the root canal (p>0.05). It is reasonable to conclude that penetration of bioceramic and epoxy resin based sealers occurred unimpressively. The type of instrument used to activate bioceramic sealer did not affect penetrability. Fluo 3 should be recommended as the fluorophore to evaluate dentinal tubule penetration of bioceramic sealers.

Resumo Este estudo objetivou avaliar a penetração nos túbulos dentinários de um novo cimento biocerâmico utilizando microscopia confocal de varredura a laser (MCVL). Um fluoróforo específico (Fluo-3) foi misturado com o cimento. Quarenta raízes distovestibulares de molares superiores foram selecionados e o preparo do canal radicular foi realizado com instrumentos Wave One Gold #35.06. As raízes foram divididas randomicamente em quatro grupos de acordo com os procedimentos obturadores: Bioceramic/Lentulo: cimento biocerâmico (Sealer Plus BC); Bioceramic/EasyClean: três ativações do cimento com instrumento Easy Clean (3 x 20 s); Bioceramic/Irrisonic: ativação ultrassônica do cimento por 30 s e AHplus/Lentulo: cimento à base de resina epóxica (AH Plus) foi utilizado com o mesmo protocolo que o grupo Bioceramic/Lentulo. Após 72h, os espécimes foram seccionados transversalmente em 2 e 7 mm do ápice radicular e analisados com MCVL. A área de penetração nos túbulos dentinários foi mensurada com Adobe Photoshop CC2018. O teste de Kruskal Wallis e T de Wilcoxon foram realizados. Os resultados de penetração foram similares para ambos os cimentos independentemente de qual técnica foi utilizada para ativá-los no interior do canal radicular (p>0,05). É razoável concluir que a penetração de cimentos biocerâmicos e à base de resina epóxica ocorreram de forma pouco expressiva. O tipo de ativação do cimento biocerâmcio não afeta sua penetrabilidade nos túbulos dentinários. Fluo 3 deve ser recomendado como o fluoróforo para avaliar a penetração intratubular de cimentos biocerâmicos.

A self-controlled study on corneal nerve invasion phenomenon of Bowman's membrane in patients with ⅢA lattice corneal dystrophy / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To analyze and study the corneal nerve invasion phenomenon of Bowman's membrane in patients with IIIA lattice corneal dystrophy by confocal laser scanning microscopy. Quantitative analysis of 10a continuous observation image data was performed, followed by self-control studies.<p>METHODS: A total of 10 patients(13 eyes)with IIIA Lattice corneal dystrophy were continuously examined by confocal laser scanning microscopy. The data were observed and analyzed. <p>RESULTS: The normal corneal nerve of Bowman's membrane(Grade 0)of IIIA LCD patients gradually decreased with the prolongation of observation time. The nerves of grade I to V involved(amyloid-wrapped nerve fibers)gradually increased correspondingly. So suggested that the corneal nerve invasion of Bowman's membrane(amyloid deposits)in patients with IIIA LCD were gradually increasing with time. <p>CONCLUSION: In patients with type ⅢA lattice corneal dystrophy, there is a neurotropic phenomenon in Bowman's membrane, which gradually worsens with the aggravation of the lesion. This lesion can explain the recurrent epithelial damage of the IIIA LCD from some degrees. Continuous observation of patients with type IIIA LCD by corneal laser confocal microscopy can well understand the development of the lesion and explain its clinical manifestations.

Influence of root canal filling techniques on sealer penetration and gutta percha/sealer ratio: a confocal laser scanning microscopy analysis / Influência das técnicas de obturação do canal radicular na penetração do cimento e na relação guta-percha / cimento: uma análise por microscopia de varredura confocal a laser

Objective: The influence of four root canal filling techniques on the penetration of an endodontic sealer into dentinal tubules and the gutta percha/ sealer ratio (GP/SR) in root canals was evaluated using confocal laser scanning microscopy (CLSM). Material and Methods: Roots of the maxillary central incisors (n=40) were prepared with ProTaper Universal files up to file F5 and assigned to five groups: continuous wave condensation, lateral condensation, single cone, Thermafill®, and negative control group. After root canal filling with gutta-percha and AH26, along with the addition of 0.01% fluorescein, the roots were cut into 2-mm slices. Using CLSM, the specimens were transversely sectioned at 3, 6, and 10 mm from the apex. Results: Sealer penetration was deeper and more frequent at 10 mm than at the 6mm and 3mm for all obturation technique. Penetration was not significantly affected by obturation techniques except single master cone tecnique. Single cone technique demonstrated the lowest sealer penetration at all levels. However, sealer thickness was strongly dependent on obturation technique. Termafill® demostrated superior GP ratio followed by continuous wave condensation, lateral condensation and single cone. Conclusion: In conclusion, the single cone technique resulted in lower sealer penetration than the other techniques, which did not differ significantly from each other. However, sealer thickness was strongly dependent on obturation technique. Termafill® demostrated superior GP ratio followed by continuous wave condensation, lateral condensation and single cone. (AU)

Objetivo: A influência de quatro técnicas de obturação do canal radicular na penetração de um cimento endodôntico nos túbulos dentinários e a relação gutapercha / cimento (GP / CIM) em canais radiculares foram avaliadas por microscopia de varredura confocal a laser (MVCL). Material e Métodos: As raízes de incisivos centrais superiores (n = 40) foram preparadas com limas ProTaper Universal até a lima F5 e distribuídas em cinco grupos: condensação de onda contínua, condensação lateral, cone único, Thermafill® e grupo de controle negativo. Após o preenchimento do canal radicular com guta-percha e AH26, juntamente com a adição de 0,01% de fluoresceína, as raízes foram cortadas em fatias de 2 mm. Usando MVCL, as amostras foram seccionadas transversalmente a 3, 6 e 10 mm do ápice. Resultados: A penetração do cimento foi mais profunda e mais frequente em 10 mm do que nos 6 mm e 3 mm para todas as técnicas de obturação. A penetração não foi significativamente afetada pelas técnicas de obturação, exceto pela técnica de cone mestre único. A técnica de cone único demonstrou a menor penetração do cimento em todos os níveis. No entanto, a espessura do cimento foi fortemente dependente da técnica de obturação. O Termafill® demonstrou uma relação superior de GP, seguida por condensação de onda contínua, condensação lateral e cone único. Conclusão: Em conclusão, a técnica de cone único resultou em menor penetração do cimento do que as outras técnicas, que não diferiram significativamente uma da outra. No entanto, a espessura do cimento foi fortemente dependente da técnica de obturação. O Termafill® demonstrou uma relação superior de GP, seguida por condensação contínua das ondas, condensação lateral e cone único. (AU)

Structural organization of dental biofilm formed in situ in the presence of sucrose associated to maltodextrin

Abstract Maltodextrins, derived from corn starch, have been added to industrialized food combined with sucrose. However it is not clear the diffusion properties of the dental biofilm matrix and the tridimensional structure of multispecies biofilms formed in the presence of these carbohydrates. Therefore, the aim of study was to investigate by confocal laser scanning microscopy (CLSM) the structural organization of the multispecies dental biofilm formed in situ under exposure to sucrose associated to maltodextrin. Adult volunteers wore an intraoral palatal appliance containing bovine enamel blocks. They were instructed to remove the appliance 8 times per day and drop the following solutions on the enamel blocks: deionized distilled water (DDW), maltodextrin, sucrose + maltodextrin or sucrose. Biofilms formed were stained and the percentage of extracellular polysaccharide (%EPS) and thickness were determined by CLSM. Biofilm formed in the presence of sucrose and sucrose + maltodextrin presented similar %EPS and higher than DDW and maltodextrin. Regarding to the biofilm thickness, sucrose and sucrose + maltodextrin treatments were thicker than DDW and maltodextrin and similar between them. The structural organization of the multispecies dental biofilm formed in situ in the presence of sucrose does not change when this carbohydrate is associated to maltodextrin.

Resumo Maltodextrinas, derivadas do amido de milho, tem sido adicionadas a alimentos industrializados combinadas à sacarose. Entretanto não estão esclarecidas as propriedades de difusão na matriz do biofilme dental e a estrutura tridimensional de biofilmes multiespécies formados na presença destes carboidratos. Portanto o objetivo deste estudo foi avaliar, através da microscopia confocal de escaneamento a laser (MCEL), a organização estrutural do biofilme dentário multiespécie formado in situ exposto à sacarose associada a maltodextrina. Voluntários adultos utilizaram dispositivos intraorais palatinos contendo blocos de esmalte bovino. Eles foram instruídos a remover os dispositivos 8 vezes por dia e gotejar as seguintes soluções sobre os blocos de esmalte: água destilada e deionizada (ADD), maltodextrina, sacarose+maltodextrina ou sacarose. Os biofilmes formados foram corados e o percentual de polissacarídeos extracelulares (%PEC) e suas espessuras foram determinados através da MCEL. Os biofilmes formados na presença de sacarose e sacarose+maltodextrina apresentaram os %PEC similares entre si, entretanto maiores do que os grupos submetidos a ADD e maltodextrina. Em relação à espessura do biofilme formado, os tratamentos sacarose e sacarose+maltodextrina apresentaram espessuras similares entre si, e maiores quando comparados aos grupos ADD e maltodextrina. A organização estrutural do biofilme dentário multiespécie formado in situ na presença de sacarose não é alterado quando este carbiodrato é associado a maltodextrina.

Application of Autofluorescence for Confocal Microscopy to Aid in Archaeoparasitological Analyses

Confocal laser scanning microscopy (CLSM) was used to examine archaeoparasitological specimens from coprolites associated with La Cueva de los Muertos Chiquitos (CMC) located near present-day Durango, Mexico. The eggs for 4 different types of parasites recovered from CMC coprolites were imaged using CLSM to assist with identification efforts. While some of the parasite eggs recovered from CMC coprolites were readily identified using standard light microscopy (LM), CLSM provided useful data for more challenging identifications by highlighting subtle morphological features and enhancing visualization of parasite egg anatomy. While other advanced microscopy techniques, such as scanning electron microscopy (SEM), may also detect cryptic identifying characters, CLSM is less destructive to the specimens. Utilizing CLSM allows for subsequent examinations, such as molecular analyses, that cannot be performed following SEM sample preparation and imaging. Furthermore, CLSM detects intrinsic autofluorescence molecules, making improved identification independent of resource and time-intensive protocols. These aspects of CLSM make it an excellent method for assisting in taxonomic identification and for acquiring more detailed images of archaeoparasitological specimens.

Clinical effect of fire needle in treating vitiligo and its CLSM image characteristics / 重庆医学

Objective To evaluate the clinical effect of fire needle in treating vitiligo and the characteristics of vitiligo image by the confocal laser scanning microscope(CLSM).Methods The randomized self-controlled experiment design was adopted.Each patient selected two symmetric or adjacent white patches and randomly received the fire needle treatment or tacrolimus treatment. The duration of treatment was 3 months.The CLSM images of white patches were recorded before treatment and after 3,6 times of fire needle treatment.Results Among 41 cases of stable stage vitiligo,The effective rates of the fire needle group and tacrolimus group were 82.9% and 78.0% respectively,and the difference was not statistically significant(P>0.05).After fire needle treat-ment,dentritic melanin cells appeared,the pigment granules gradually appeared around the basal layer and corpora papillare,and formed the pigment ring.Conclusion Fire needle and tacrolimus have the similar effect in treating vitiligo,moreover CLSM can be used as the non-invasive,objective and reliable detection means of the recovery of vitiligo melanocyte.

Improved dentin disinfection by combining different-geometry rotary nickel-titanium files in preparing root canals

OBJECTIVES: This study was to evaluate the antibacterial effect of different instrumentation and irrigation techniques using confocal laser scanning microscopy (CLSM) after root canal inoculation with Enterococcus faecalis (E. faecalis). MATERIALS AND METHODS: Mesiobuccal and mesiolingual canals of extracted mandibular molars were apically enlarged up to a size 25 hand K-file, then autoclaved and inoculated with E. faecalis. The samples were randomly divided into 4 main groups according to the system of instrumentation and irrigation: an XP-endo Shaper (XPS) combined with conventional irrigation (XPS/C) or an XP-endo Finisher (XPF) (XPS/XPF), and iRaCe combined with conventional irrigation (iRaCe/C) or combined with an XPF (iRaCe/XPF). A middle-third sample was taken from each group, and then the bacterial reduction was evaluated using CLSM at a depth of 50 µm inside the dentinal tubules. The ratio of red fluorescence (dead cells) to green-and-red fluorescence (live and dead cells) represented the percentage of bacterial reduction. The data were then statistically analyzed using the Kruskal-Wallis test for comparisons across the groups and the Dunn test was used for pairwise comparisons. RESULTS: The instrumentation and irrigation techniques had a significant effect on bacterial reduction (p < 0.05). The iRaCe/XPF group showed the strongest effect, followed by the XPS/XPF and XPS/C group, while the iRaCe/C group had the weakest effect. CONCLUSIONS: Combining iRaCe with XPF improved its bacterial reduction effect, while combining XPS with XPF did not yield a significant improvement in its ability to reduce bacteria at a depth of 50 µm in the dentinal tubules.

Influence of Root Canal Filling Techniques on Sealer Penetration and Bond Strength to Dentin

Abstract The influence of root canal filling techniques on the penetration of AH Plus into dentinal tubules was evaluated using confocal laser scanning microscopy (CLSM) and the impact on the bond strength of the filling material on the root dentin was evaluated using the push-out test. Roots of the maxillary central incisors (n=40) were prepared with ProTaper Universal and assigned to four groups: lateral condensation, vertical compaction, modified Tagger hybrid and single cone. After root canal filling with gutta-percha and AH Plus, along with the addition of 0.01% fluorescein, the roots were cut into 2-mm slices. CLSM was performed on the first slice of each root third, followed by the push-out test. The second slice was analyzed under SEM. ANOVA demonstrated a statistically significant difference between the root canal filling techniques and the root thirds (p<0.05). For percent penetration, Tukey's test showed that the vertical compaction (91.77±13.51), lateral condensation (89.18±10.91) and modified Tagger hybrid (87.53±15.42) presented the highest values, which significantly differed (p<0.05) from those of the single cone (78.19±22.46). For bond strength (MPa), lateral condensation (3.42±0.54) and vertical compaction (3.47±0.53) had the highest values (p<0.05), while the modified Tagger hybrid (2.19±0.41) and single cone (0.54±0.21) had the lowest. In conclusion, the lateral condensation, vertical compaction and Modified Tagger's hybrid techniques resulted in greater penetration and bond strength to the intra-radicular dentin.

Resumo Avaliou-se a influência da técnica obturadora na penetração do cimento AH Plus nos túbulos dentinários, por meio de microscopia confocal de varredura a laser (MCVL), e o impacto na resistência de união do material obturador à dentina, por meio do teste de push out. Raízes de incisivos centrais superiores (n=40) foram preparadas com ProTaper Universal e distribuídas em 4 grupos: Condensação lateral, Compactação vertical, Híbrida de Tagger Modificada e Cone único. Após obturação com guta percha e AH Plus acrescido de fluoresceína 0,01%, as raízes foram seccionadas em slices de 2,0 mm. O primeiro slice de cada terço foi submetido à MCVL e, posteriormente, ao teste de push out. O segundo slice foi analisado em MEV. ANOVA demonstrou diferença estatisticamente significante para as técnicas obturadoras e terços radiculares (p<0,05). Para o percentual de penetração, Tukey evidenciou que compactação vertical (91,77±13,51), Condensação lateral (89,18±10,91) e Híbrida de Tagger Modificada (87,53±15,42) apresentaram os maiores valores, estatisticamente diferentes (p<0,05) da Cone único (78,19±22,46). Na resistência de união (MPa), Condensação lateral (3,42±0,54) e Compactação vertical (3,47±0,53) obtiveram os maiores valores, diferentes (p<0,05) da Híbrida de Tagger Modificada (2,19±0,41) e da Cone único (0,54±0,21), que teve os menores valores. Concluiu-se que as técnicas obturadoras Condensação lateral, Compactação vertical e Híbrida de Tagger Modificada propiciaram maior penetração e resistência de união à dentina intrarradicular.

Methods of Hematoxylin and Erosin Image Information Acquisition and Optimization in Confocal Microscopy / 대한의료정보학회지

OBJECTIVES: We produced hematoxylin and eosin (H&E) staining-like color images by using confocal laser scanning microscopy (CLSM), which can obtain the same or more information in comparison to conventional tissue staining. METHODS: We improved images by using several image converting techniques, including morphological methods, color space conversion methods, and segmentation methods. RESULTS: An image obtained after image processing showed coloring very similar to that in images produced by H&E staining, and it is advantageous to conduct analysis through fluorescent dye imaging and microscopy rather than analysis based on single microscopic imaging. CONCLUSIONS: The colors used in CLSM are different from those seen in H&E staining, which is the method most widely used for pathologic diagnosis and is familiar to pathologists. Computer technology can facilitate the conversion of images by CLSM to be very similar to H&E staining images. We believe that the technique used in this study has great potential for application in clinical tissue analysis.

Antifungal effects of synthetic human β-defensin 3-C15 peptide

OBJECTIVES: The purpose of this ex vivo study was to compare the antifungal activity of a synthetic peptide consisting of 15 amino acids at the C-terminus of human β-defensin 3 (HBD3-C15) with calcium hydroxide (CH) and Nystatin (Nys) against Candida albicans (C. albicans) biofilm. MATERIALS AND METHODS: C. albicans were grown on cover glass bottom dishes or human dentin disks for 48 hr, and then treated with HBD3-C15 (0, 12.5, 25, 50, 100, 150, 200, and 300 µg/mL), CH (100 µg/mL), and Nys (20 µg/mL) for 7 days at 37℃. On cover glass, live and dead cells in the biomass were measured by the FilmTracer Biofilm viability assay, and observed by confocal laser scanning microscopy (CLSM). On dentin, normal, diminished and ruptured cells were observed by field-emission scanning electron microscopy (FE-SEM). The results were subjected to a two-tailed t-test, a one way analysis variance and a post hoc test at a significance level of p = 0.05. RESULTS: C. albicans survival on dentin was inhibited by HBD3-C15 in a dose-dependent manner. There were fewer aggregations of C. albicans in the groups of Nys and HBD3-C15 (≥ 100 µg/mL). CLSM showed C. albicans survival was reduced by HBD3-C15 in a dose dependent manner. Nys and HBD3-C15 (≥ 100 µg/mL) showed significant fungicidal activity compared to CH group (p < 0.05). CONCLUSIONS: Synthetic HBD3-C15 peptide (≥ 100 µg/mL) and Nys exhibited significantly higher antifungal activity than CH against C. albicans by inhibiting cell survival and biofilm.

Clinical observation of the efficacy of No. 1 Baiban granules on vitiligo / 天津医药

Objective To observe the efficacy of No. 1 Baiban granules in the treatment of vitiligo using skin confocal laser scanning microscopy. Methods The study was approved by the hospital ethics committee. Nine-two patients with qi stagnation and blood stasis were collected from Department of Dermatology of the Affiliated Hospital of Tianjin Academy of Traditional Chinese Medicine and were randomly assigned to two groups. The experimental group (n=49) received oral Baiban granuleΙ, 3 g, twice a day, transfer factor capsules 6 mg, 3 times a day, 0.1%mometasone furoate cream (excluding the face) for external use, once a day, or 0.1%tacrolimus ointment (Afacial), twice a day for 3 months. The control group (n=43) only received oral transfer factor capsules 6 mg, 3 times a day, 0.1%mometasone furoate cream (except for the face) for external use, once a day, or 0.1%tacrolimus ointment (face), twice a day, treatment for 3 months. The therapeutic effect was observed in two groups. CLSM was used to monitor the compound color and pigment recovery level, and changes of the number of dendritic cells and inflammatory cell infiltration in skin. Results The compound color was improved significantly after treatment in experimental group and control group. The total effective rate was higher in experimental group than that of control group (81.63% vs. 69.77%,χ2=3.947,P<0.05). Results of CLSM observation showed that the pigment recovery of skin lesion, the proportion of dendritic cells and inflammatory cells were significantly higher in experimental group than those of control group (P < 0.05). Conclusion The treatment effect of No. 1 Baiban granules is more obvious for vitiligo of qi stagnation and blood stasis.

Anti-biofilm efficacy of silver nanoparticles against MRSA and MRSE isolated from wounds in a tertiary care hospital.

Purpose: Different approaches have been used for preventing biofi lm-related infections in health care settings. Many of these methods have their own de-merits, which include chemical-based complications; emergent antibiotic resistant strains, etc. The formation of biofi lm is the hallmark characteristic of Staphylococcus aureus and S. epidermidis infection, which consists of multiple layers of bacteria encased within an exopolysachharide glycocalyx. Nanotechnology may provide the answer to penetrate such biofi lms and reduce biofi lm formation. Therefore, the aim of present study was to demonstrate the biofi lm formation by methicillin resistance S. aureus (MRSA) and methicillin resistance S. epidermidis (MRSE) isolated from wounds by direct visualisation applying tissue culture plate, tube and Congo Red Agar methods. Materials and Methods: The anti-biofi lm activity of AgNPs was investigated by Congo Red, scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) techniques. Results: The minimum inhibitory concentration (MIC) was found to be in the range of 11.25-45 μg/ml. The AgNPs coated surfaces effectively restricted biofi lm formation of the tested bacteria. Double fl uorescent staining (propidium iodide staining to detect bacterial cells and fl uorescein isothiocyanate concanavalin A (Con A-FITC) staining to detect the exopolysachharides matrix) technique using CLSM provides the visual evidence that AgNPs arrested the bacterial growth and prevent the glycocalyx formation. In our study, we could demonstrate the complete anti-biofi lm activity AgNPs at a concentration as low as 50 μg/ml. Conclusions: Our fi ndings suggested that AgNPs can be exploited towards the development of potential anti-bacterial coatings for various biomedical and environmental applications. In the near future, the AgNPs may play major role in the coating of medical devices and treatment of infections caused due to highly antibiotic resistant biofilm.

Influence of the Method in Root Canal Filling Using Active Lateral Compaction Techniques

The aim of this study was to evaluate, by stereomicroscopy and confocal laser microscopy, the influence of different lateral compaction methods for the obturation quality, as well as the time spent for the procedure. Thirty root canals of freshly extracted single-rooted human teeth were prepared with the ProTaper system up to F5 instrument and filled with gutta-percha and AH Plus sealer by the lateral compaction technique. The teeth were divided into 3 groups (n=10) in accordance with the method used for the lateral compaction, as follows: Manual, Mechanical and Ultrasonic. The sealers were stained with rhodamine B dye in a proportion of 0.1% per gram in weight to allow for the analysis under a confocal microscope. During the root filling procedure, the time spent was recorded with a stopwatch. The specimens were stored at 37 °C for 48 h, and then sectioned at 2, 4 and 6 mm from the apex. The percentage of gutta-percha, sealer and void areas were evaluated using a stereomicroscope and sealer penetration perimeter by confocal laser microscope. Statistical analyses were performed using the Kruskall-Wallis and Dunn tests (p<0.05). The mechanical method required a shorter time for the lateral compaction than the manual method (p<0.05). There were no significant differences (p>0.05) for the perimeter of the dentin with sealer penetration among all groups. The mechanical method showed a higher (p<0.05) percentage of gutta-percha and less sealer at the 4 mm section in comparison with the manual method. The ultrasonic group showed intermediate values. The void areas found in the root filling were similar (p>0.05) between the three methods. In conclusion, the fastest lateral compaction was achieved with the mechanical method, and all the methods showed void areas in the root filling.

O objetivo deste estudo foi avaliar a influência de diferentes técnicas de condensação lateral em relação a qualidade da obturação por meio de estereomicroscopia e microscopia confocal, bem com o tempo gasto para realizar o referido procedimento. Trinta canais radiculares de dentes humanos unirradiculares foram modelados com o sistema ProTaper, finalizando com o instrumento F5. Em seguida os canais foram obturados com guta percha e cimento AH Plus por meio da técnica de condensação lateral. Os espécimes foram divididos em 3 grupos (n=10) de acordo com o método de condensação lateral utilizado, sendo: Manual, Mecânico, Ultrassônico. Os cimentos foram manipulados com rodamina B na proporção de 0.1% de peso para permitir a análise com microscopia confocal. O tempo gasto para realizar cada procedimento de obturação foi registrado com o uso de um cronômetro. Os espécimes foram armazenados a 37 °C durante 48 h, para em seguida serem seccionados a 2, 4 e 6 mm a partir do ápice radicular. A porcentagem da área de guta percha, cimento e vazios foram avaliados com estereomicroscópio. A penetração de cimento foi avaliada por meio de microscópio confocal. A análise estatística foi realizada utilizando os testes de Kruskall-Wallis e Dunn (P < 0.05). O método mecânico exigiu menos tempo para realizar a condensação lateral em comparação com o método manual (p<0.05). Não foi encontrada diferença (p>0.05) em relação ao perímetro de penetração de cimento nos túbulos dentinários entre todos os grupos avaliados. O método mecânico apresentou maior (p<0.05) porcentagem de guta percha e menos cimento no nível de 4 mm quando comparado com o método manual. O grupo do ultrassom apresentou resultados intermediários. Áreas de vazios encontradas foram similares (p>0.05) entre os grupos. Foi possível concluir que o método mecânico foi mais rápido para a técnica de condensação lateral, mas todos os métodos de obturação estudados apresentaram espaços vazios na massa obturadora.

Effect of subinhibitory concentrations of fluoroquinolones on biofilm production by clinical isolates of Streptococcus pyogenes.

Background & objectives: Subinhibitory concentrations (sub-MICs) of antibiotics, although not able to kill bacteria, but influence bacterial virulence significantly. Fluoroquinolones (FQs) which are used against other bacterial pathogens creates resistance in non-targeted Streptococcus pyogenes. This study was undertaken to characterize the effect of sub-MICs of FQs on S. pyogenes biofilm formation. Methods: Biofilm forming six M serotypes M56, st38, M89, M65, M100 and M74 of S. pyogenes clinical isolates were challenged against four FQs namely, ciprofloxacin, ofloxacin, levofloxacin and norfloxacin. The antibiofilm potential of these FQs was analysed at their subinhibitory concentrations (1/2 to 1/64 MIC) using biofilm assay, XTT reduction assay, scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Results: Among the four FQs tested, ofloxacin and levofloxacin at 1/2 MIC showed the maximum inhibition (92%) of biofilm formation against M56 and M74 serotypes. FQs effectively interfered in the microcolony formation of S. pyogenes isolates at 1/2 to 1/8 sub-MICs. Inhibition of biofilm formation was greatly reduced beyond 1/16 MICs and allowed biofilm formation. XTT reduction assay revealed the increase in metabolic activity of S. pyogenes biofilm against the decrease in FQs concentration. SEM and CLSM validated the potential of sub-MICs of FQs against the six S. pyogenes. Interpretation & conclusions: Our results showed that the inhibitory effect all four FQs on S. pyogenes biofilm formation was concentration dependent. FQs at proper dosage can be effective against S. pyogenes and lower concentrations may allow the bacteria to form barriers against the antibiotic in the form of biofilm.

Clinical evaluation of rapid diagnosis of fungal keratitis by the combination of corneal scraping with laser scanning confocal microscopy / 中华实验眼科杂志

Background The rapid diagnosis can win more treating opportunities for patients with fungal keratitis.Even though the fungal culture is the gold standard for the diagnosis of fungal keratitis,it is difficult in early diagnosis due to the long duration of cultivation and false-negative rate.Objective This trial was to explore the clinical value in the rapid diagnosis of fungal keratitis by the combination of corneal scraping with laser scanning confocal microscopy.Methods Corneal scraping and laser scanning confocal microscopy were separately performed in 167 eyes of 167 patients with fungal keratitis.All the eyes were examined by the slit lamp,followed by laser scanning confocal microscope,and then the 10％ KOH corneal smear was examined under the optical microscope.Results The positive rate of diagnosis was 75％ (125/167) by corneal scraping,and that by laser scanning confocal microscopy was 91％ (152/167).The positive rate of examining outcome was significantly higher in laser scanning confocal microscopy than that of corneal scraping (x2 =14.88,P =0.00).The positive results were 114 cases and negative results were 4 cases by two methods,with the concordance rate 70.7％ (118/167).The hyphae or spore were seen in 32 cases by laser scanning confocal microscopy in 42 negative cases by corneal scraping,and in 15 negative cases by confocal laser scanning microscopy,11 positive outcomes were offered by corneal scraping.Conclusions The combined application of corneal scraping with confocal laser scanning microscopy can improve and speed up the diagnosis positive rate of fungal keratitis.

Study of the effective of efflux pump inhibitors in photodynamic therapy inhibition the cariogenic bacteria in dental plaque biofilms / 国际生物医学工程杂志

Objective To Study the effect of efflux pump-inhibitors(EPI)-Verapamil in photodynamic therapy (PDT) using hematoporphyrin monomethylether (HMME) as photosensitizer on the cariogenic bacteria in dental plaque biofilms.Methods According to the administrator order of the verapamil and photosensitizer in PDT,streptococcus mutans,streptococcus sanguis,eosinophilic lactobacillus and actinomyces viscosus were used to establish the dental plaque biofilm model.The experiment was divided into five groups,group A was incubated with the photosensitizer and verapamil group,group B using verapamil before incubated the photosensitize,group C suing photosensitizer before incubated verapamil,group D with PDT only,group E was control group.After laser treatment,the influence of the dental plaque biofilms was observed by confocal laser scanning microscope.Results As saline-treated group is a group of normal the dental plaque biofilms.In PDT only group,compared with the saline group,red fluorescence increased significantly,the bacteria lose accumulation capacity,and were isolated and scattered in dispersed state.In PDT plus verapamil group,compared with only PDT group,green staining increased,bacterial activity increased.In group B,cells were incubated with verapamil before incubated the photosensitizer group,green staining increased significantly,red fluorescence reduced,indicating live bacteria increased,and bacteria activity was significantly increased.Conclusion PDT is an effective method in eliminating cariogenic bacteria of dental plaque biofilms.Bacterial efflux pump inhibitors can lower HMME-PDT inhibition cariogenic bacteria in dental plaque biofilm,and pre-verapamil administration could significantly inhibit the effect of PDT treatment of dental caries.

In vitro biofilm forming potential of Streptococcus suis isolated from human and swine in China

Streptococcus suis is a swine pathogen and also a zoonotic agent. The formation of biofilms allows S. suis to become persistent colonizers and resist clearance by the host immune system and antibiotics. In this study, biofilm forming potentials of various S. suis strains were characterized by confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM) and tissue culture plates stained with crystal violet. In addition, the effects of five antimicrobial agents on biofilm formation were assayed in this study. S. suis produced biofilms on smooth and rough surface. The nutritional contents including glucose and NaCl in the growth medium modulated biofilm formation. There was a significant difference in their biofilm-forming ability among all 46 S. suis strains. The biofilm-forming potential of S. suis serotype 9 was stronger than type 2 and all other types. However, biofilm formation was inhibited by five commonly used antimicrobial agents, penicillin, erythromycin, azithromycin, ciprofloxacin, and ofloxacin at subinhibitory concentrations, among which inhibition of ciprofloxacin and ofloxacin was stronger than that of other three antimicrobial agents.Our study provides a detailed analysis of biofilm formation potential in S. suis, which is a step towards understanding its role in pathogenesis, and eventually lead to a better understanding of how to eradicate S. suis growing as biofilms with antibiotic therapy.