QUANTIFICATION OF RETINAL CAPILLARY NONPERFUSION IN DIABETICS USING WIDE-FIELD OPTICAL COHERENCE TOMOGRAPHY ANGIOGRAPHY.

PURPOSE: To combine advances in high-speed, wide-field optical coherence tomography angiography (OCTA) with image processing methods for semiautomatic quantitative analysis of capillary nonperfusion in patients with diabetic retinopathy (DR). METHODS: Sixty-eight diabetic patients (73 eyes), either without retinopathy or with different degrees of retinopathy, were prospectively recruited for volumetric swept-source OCTA imaging using 12 mm × 12 mm fields centered at the fovea. A custom, semiautomatic software algorithm was used to quantify areas of capillary nonperfusion. RESULTS: The mean percentage of nonperfused area was 0.1% (95% confidence interval: 0.0-0.4) in the eyes without DR; 2.1% (95% confidence interval: 1.2-3.7) in the nonproliferative DR eyes (mild, moderate, and severe), and 8.5% (95% confidence interval: 5.0-14.3) in the proliferative DR eyes. The percentage of nonperfused area increased in a statistically significant manner from eyes without DR, to eyes with nonproliferative DR, to eyes with proliferative DR. CONCLUSION: Capillary nonperfusion area in the posterior retina increases with increasing DR severity as measured by swept-source OCTA. Quantitative analysis of retinal nonperfusion on wide-field OCTA may be useful for early detection and monitoring of disease in patients with diabetes and DR.

Optical coherence tomography to evaluate the effects of oxidative hair dye on the fiber.

BACKGROUND/PURPOSE: Oxidative hair dyes can damage the hair, since these chemical procedures are involved to change the fiber structure and therefore changes in their mechanical and surface properties. Evaluate and compare the effect of the two colors of oxidative hair dye emulsions on Caucasian hair. This research analyzed the Dark brown hair untreated (I); Dark brown hair treated with light brown dye (II); Dark brown hair treated with light blond dye (III); Light blond hair untreated (IV); Light blond hair treated with light brown dye (V); Light blond hair treated with light blond dye (VI) on Caucasian hair. METHODS: The hair samples were submitted to breaking strength, color, and optical coherence tomography (OCT) analysis. RESULTS: For the breaking strength assay no presented statistically significant differences between treatments. The parameters of color and brightness can differ in some hair dye formulations, but also the hair type can respond differently. The OCT images of the sample I and IV was possible observed, clearly Medulla and Cortex, which was not observed clearly after treatment with both oxidative hair dye colors. CONCLUSION: Based on the results, the oxidative hair dyes increased alteration in color and ultrastructure of hair.

Obtaining artificially caries-affected dentin for in vitro studies.

AIM: This study evaluated and improved a protocol for obtaining standard caries-affected dentin (CAD) by Streptococcus mutans biofilm demineralization process. MATERIALS AND METHODS: Forty-eight human molars were divided in six experimental groups, according to: period of cariogenic challenge (7, 14 or 21 days) and type of dentin (erupted or unerupted teeth). After complete cariogenic challenge sound and CAD dentin were evaluated by: visual inspection (VI), digital radiography (DR), optical coherence tomography (OCT) and laser fluorescence (LF). RESULTS: Visual inspection confirmed the formation of CAD based on tissue yellowing and loss of surface gloss. Digital radiography detected the presence of radiolucent images, suggesting caries. Three calibrated examiners viewed all images obtained by VI and DR and were able to distinguish healthy from CAD. Fisher's exact statistical test (p < 0.05) confirmed no difference between groups by VI (G1/G4: p = 0.6; G2/G5: p = 1; G3/G6: p = 1) or DR (G1/G4: p = 1; G2/G5: p = 1; G3/G6: p = 1). Both LF values and demineralization depth, as determined by OCT, were subjected to ANOVA (p < 0.05). For LF, a statistically significant difference was observed for the type of substrate (p = 0.001). For OCT, no statistically significant differences in the type of substrate (p = 0.163), length of cariogenic challenge (p = 0.512) or interaction between factors (p = 0.148) were observed. Scanning electron micrographs confirmed the presence of CAD; a more uniform demineralization surface was observed in the dentin of unerupted teeth. CONCLUSION: This protocol suggests that standard CAD can be obtained in 7 days of cariogenic challenge using unerupted teeth. Clinical significance: With the new perspective on the clinical treatment of caries lesions, bonding is increasingly performed to demineralize CAD, which is susceptible to remineralization. A useful protocol to standardize the production of CAD, by microbiological cariogenic challenge, would be an important contribution to laboratorial test in the field of operative dentistry.

Role of non-carious cervical lesions multicausality in the behavior of respective restorations.

OBJECTIVES: To evaluate biocorrosion and eccentric occlusal loading interplay in marginal quality of cervical restorations. METHODS: Cervical wedge-shaped cavities were prepared in extracted premolars and restored with a composite. Premolars underwent either an erosive challenge (E: 1% citric acid/10 min), eccentric occlusal loading (EOL: 150 N/2.5 Hz/106 cycles), E before EOL (E + EOL), E intermediate to EOL (EOL/E/EOL), E after EOL (EOL + E), or no E or EOL (C: control). Marginal quality was analyzed based on a series of Optical Coherence Tomography images. Each of the margins was assigned a gap score (0, 1, 2, or 3) and measurement (µm). For each margin, scores data were analyzed with Kruskall Wallis and Dunn tests, and µm data, with Kruskall Wallis. Overall and for each group, the different margins were compared using Wilcoxon signed-rank test, and the correlation between scores and µm, Spearman's correlation coefficient (α = 0.05). RESULTS: E and EOL, even if associated, did not influence enamel marginal quality. EOL/E/EOL impaired dentin/cementum marginal quality only in the case of scores and compared to E. E + EOL, EOL + E or EOL and even C, without differences between each other, did not influence results differently from E or EOL/E/EOL. Margins in dentin/cementum always showed lengthier gaps. Except for C, E and EOL + E cervical margin, there was a strong positive correlation between scores and µm. CONCLUSIONS: Eccentric occlusal loading and/or biocorrosion cannot be assumed as causes of marginal failure of cervical restorations in wedge-shaped cavities. A relevant concern may still be the establishment of adhesive interfaces in dentin/cementum. CLINICAL SIGNIFICANCE: Although non-carious cervical lesions are strongly being recognized multifactorial and their respective restorations not always behave as expected, biocorrosion and eccentric occlusal loading interplay cannot serve as an explanation for marginal gaps they often present.

Analysis of enamel/restoration interface submitted cariogenic challenge and fluoride release.

The treatment of high-risk patients still is a challenge. The understanding and development non-invasive, non-destructive, and non-ionizing techniques, can help to guide the treatment and the diagnosis of primary and recurrent caries. The present study evaluated the behavior of enamel/restoration interface after a cariogenic challenge by Fourier domain optical coherence tomography (FD-OCT), scanning electron microscopy (SEM) examination, and the fluoride release of the different restorative materials. Cavities (1.5 × 0.5 mm) were performed in enamel surface and divided into groups (n = 8): glass ionomer cement (GIC), resin-modified glass ionomer cement (RMGIC), and resin composite (RC). The samples were submitted to pH-cycling, and the solutions analyzed for cumulative fluoride by ion-analyzer. The morphology was analyzed by SEM through replicas. The optical attenuation coefficient (OAC) was calculated through exponential decay from the images generated by FD-OCT. Data were analyzed considering α = 0.05. OAC values increased for all groups after pH-cycling indicating demineralization (p < .05). Considering the remineralizing solution, RMGIC presented higher fluoride release rate, followed by GIC, while RC did not release any fluoride. Yet for the demineralizing solution, RMGIC and GIC released similar fluoride rates, overcoming RC (p < .05). Micrographs revealed no changes on the restorations margins, although enamel detachment was observed for RC and GIC after pH-cycling.

Prospective ultramorphological characterization of human hair by optical coherence tomography.

BACKGROUND/PURPOSE: The continuous advancement in cosmetic science has led to an increasing demand for the development of non-invasive, reliable scientific techniques directed toward claim substantiation, which is of utmost relevance, to obtain data regarding the efficacy and safety of cosmetic products. METHODS: In this work, we used the optical coherence tomography (OCT) technique to produce in vitro transversal section-images of human hair. We also compared the OCT signal before and after chemical treatment with an 18% w/w ammonium thioglycolate solution. RESULTS: The mean diameter of the medulla was 29+/-7 microm and the hair diameter was 122+/-16 microm in our samples of standard Afro-ethnic hair. A three-dimensional (3D) image was constructed starting from 601 cross-sectional images (slices). Each slice was taken in steps of 6.0 microm at eight frames per second, and the entire 3D image was constructed in 60 s. CONCLUSION: It was possible to identify, using the A-scan protocol, the principal structures: the cuticle, cortex and medulla. After chemical treatment, it was not possible to identify the main structures of hair fiber due to index matching promoted by deleterious action of the chemical agent.

Controlling for Artifacts in Widefield Optical Coherence Tomography Angiography Measurements of Non-Perfusion Area.

The recent clinical adoption of optical coherence tomography (OCT) angiography (OCTA) has enabled non-invasive, volumetric visualization of ocular vasculature at micron-scale resolutions. Initially limited to 3 mm × 3 mm and 6 mm × 6 mm fields-of-view (FOV), commercial OCTA systems now offer 12 mm × 12 mm, or larger, imaging fields. While larger FOVs promise a more complete visualization of retinal disease, they also introduce new challenges to the accurate and reliable interpretation of OCTA data. In particular, because of vignetting, wide-field imaging increases occurrence of low-OCT-signal artifacts, which leads to thresholding and/or segmentation artifacts, complicating OCTA analysis. This study presents theoretical and case-based descriptions of the causes and effects of low-OCT-signal artifacts. Through these descriptions, we demonstrate that OCTA data interpretation can be ambiguous if performed without consulting corresponding OCT data. Furthermore, using wide-field non-perfusion analysis in diabetic retinopathy as a model widefield OCTA usage-case, we show how qualitative and quantitative analysis can be confounded by low-OCT-signal artifacts. Based on these results, we suggest methods and best-practices for preventing and managing low-OCT-signal artifacts, thereby reducing errors in OCTA quantitative analysis of non-perfusion and improving reproducibility. These methods promise to be especially important for longitudinal studies detecting progression and response to therapy.

Evaluation of femtosecond laser-induced breakdown spectroscopy for analysis of animal tissues.

The aim of this work was to evaluate the performance of femtosecond laser-induced breakdown spectroscopy (fs-LIBS) for the determination of elements in animal tissues. Sample pellets were prepared from certified reference materials, such as liver, kidney, muscle, hepatopancreas, and oyster, after cryogenic grinding assisted homogenization. Individual samples were placed in a two-axis computer-controlled translation stage that moved in the plane orthogonal to a beam originating from a Ti:Sapphire chirped-pulse amplification (CPA) laser system operating at 800 nm and producing a train of 840 microJ and 40 fs pulses at 90 Hz. The plasma emission was coupled into the optical fiber of a high-resolution intensified charge-coupled device (ICCD)-echelle spectrometer. Time-resolved characteristics of the laser-produced plasmas showed that the best results were obtained with delay times between 80 and 120 ns. Data obtained indicate both that it is a matrix-independent sampling process and that fs-LIBS can be used for the determination of Ca, Cu, Fe, K, Mg, Na, and P, but efforts must be made to obtain more appropriate detection limits for Al, Sr, and Zn.

Antimicrobial photodynamic therapy combined to periodontal treatment: Experimental model.

BACKGROUND: Antimicrobial photodynamic therapy (aPDT) has been used as an adjuvant treatment for periodontitis. It combines a photosensitizer with a light source to induce reactive oxygen species and kill microbial cells. PpNetNI is a protoporphyrin derivative, and it has a chemical binding site at biofilm and great affinity to microbial cells. The aim of this study was to investigate the effects of aPDT as an adjuvant treatment for periodontitis. METHODS: Thirty healthy male rats Wistar (Rattus norvegicus) were used in this study (Approved by UNINOVE Ethical committee AN0029/2015). Periodontitis was induced by placing a cotton ligature around the first mandibular molar in a subgengival position. The contralateral mandibular first molar received neither a ligature nor any treatment, and was used as a control. After 7 days, the ligature was removed and all animals received scaling and root planing (SRP) and were divided according to the following treatments: SRP group (received SRP and irrigation with PpNetNI, 10µM) and aPDT group (PpNetNI 10µM followed by LED irradiation). aPDT was performed with a LED (630nm) with an output power of 400mW (fluence-rate 200mW/cm2; fluence 18J/cm2). Rats were euthanized at 24h, 48h and 7days postoperatively. The area of bone loss in vestibular region of the first molar was evaluated by Optical Coherence Tomography (OCT, THORLABS LTD., Ely, UK). Data were analyzed statistically (ANOVA and Tukey tests, p<0.05). RESULTS: The animals treated by aPDT showed bone gain of approximately 30% compared to the SRP group following 7days from the treatment. CONCLUSION: aPDT promoted bone recovery 7days after periodontal intervention.

Evaluation of dental enamel caries assessment using Quantitative Light Induced Fluorescence and Optical Coherence Tomography.

An in vitro study of morphological alterations between sound dental structure and artificially induced white spot lesions in human teeth, was performed through the loss of fluorescence by Quantitative Light-Induced Fluorescence (QLF) and the alterations of the light attenuation coefficient by Optical Coherence Tomography (OCT). To analyze the OCT images using a commercially available system, a special algorithm was applied, whereas the QLF images were analyzed using the software available in the commercial system employed. When analyzing the sound region against white spot lesions region by QLF, a reduction in the fluorescence intensity was observed, whilst an increase of light attenuation by the OCT system occurred. Comparison of the percentage of alteration between optical properties of sound and artificial enamel caries regions showed that OCT processed images through the attenuation of light enhanced the tooth optical alterations more than fluorescence detected by QLF System. QLF versus OCT imaging of enamel caries: a photonics assessment.

Optical coherence tomography as an auxiliary tool for the screening of radiation-related caries.

OBJECTIVE: The aim of this study was to evaluate the morphological alterations of radiation-related caries using optical coherence tomography. METHODS: Thirty-six extracted teeth from 11 patients who had undergone radiotherapy were sectioned in the sagittal axis in the center of the carious lesion, and 100 µm thick sections were obtained from each specimen. One sample from each tooth was investigated by an optical coherence tomography (OCT) system, and the results were compared with histological images from polarized light microscopy. RESULTS: In OCT dentin caries images, the demineralized area appeared as a white region, whereas the translucent zone appeared as a dark area, a similar pattern also seen in coronal caries. In noncavitated enamel lesions clinically observed as brown discoloration, the area of high porosity, and also the dark color, absorbs part of the light, resulting in a dark pattern. Finally, the involvement of dentin-enamel junction (DEJ) or cement-enamel junction (CEJ) could be clearly observed, when present and marked alterations along the CEJ could be noted, as junction continuity loss, gap formation, and mineral loss tissue. CONCLUSIONS: The OCT technique was able to characterize radiation-related caries, from a morphological point of view. Also demonstrated was its potential benefit for use in the clinical monitoring of radiation-related carious process.

Alternative methods for determining shrinkage in restorative resin composites.

OBJECTIVES: The purpose of this study was to evaluate polymerization shrinkage of resin composites using a coordinate measuring machine, optical coherence tomography and a more widely known method, such as Archimedes Principle. Two null hypothesis were tested: (1) there are no differences between the materials tested; (2) there are no differences between the methods used for polymerization shrinkage measurements. METHODS: Polymerization shrinkage of seven resin-based dental composites (Filtek Z250™, Filtek Z350™, Filtek P90™/3M ESPE, Esthet-X™, TPH Spectrum™/Dentsply 4 Seasons™, Tetric Ceram™/Ivoclar-Vivadent) was measured. For coordinate measuring machine measurements, composites were applied to a cylindrical Teflon mold (7 mm × 2 mm), polymerized and removed from the mold. The difference between the volume of the mold and the volume of the specimen was calculated as a percentage. Optical coherence tomography was also used for linear shrinkage evaluations. The thickness of the specimens was measured before and after photoactivation. Polymerization shrinkage was also measured using Archimedes Principle of buoyancy (n=5). Statistical analysis of the data was performed with ANOVA and the Games-Howell test. RESULTS: The results show that polymerization shrinkage values vary with the method used. Despite numerical differences the ranking of the resins was very similar with Filtek P90 presenting the lowest shrinkage values. SIGNIFICANCE: Because of the variations in the results, reported values could only be used to compare materials within the same method. However, it is possible rank composites for polymerization shrinkage and to relate these data from different test methods. Independently of the method used, reduced polymerization shrinkage was found for silorane resin-based composite.