Anti-4-1BB×PD-L1 Bispecific Antibody Reinvigorates Tumor-Specific Exhausted CD8+ T Cells and Enhances the Efficacy of Anti-PD-1 Blockade.

PURPOSE: To overcome the limited efficacy of immune checkpoint blockade, there is a need to find novel cancer immunotherapeutic strategies for the optimal treatment of cancer. The novel anti-4-1BB×PD-L1 bispecific antibody-ABL503 (also known as TJ-L14B)-was designed to simultaneously target PD-L1 and 4-1BB, and demonstrated strong antitumor T-cell responses without considerable toxicity. Here, we investigated how the combination of ABL503 and anti-PD-1 blockade affected the reinvigoration of exhausted tumor-infiltrating CD8+ T cells (CD8+ TILs) and anti-tumor efficacy. EXPERIMENTAL DESIGN: Single cell suspensions of hepatocellular carcinoma and ovarian cancer from treatment-naive patients were used for immunophenotyping of CD8+ TILs and in vitro functional assays. Humanized hPD-1/hPD-L1/h4-1BB triple knock-in mice were used to evaluate the effects of ABL503 and anti-PD-1 blockade in vivo. RESULTS: We observed that ABL503 successfully restored the functions of 4-1BB+ exhausted CD8+ TILs, which were enriched for tumor-specific T cells but unresponsive to anti-PD-1 blockade. Importantly, compared to anti-PD-1 blockade alone, the combination of ABL503 and anti-PD-1 blockade further enhanced the functional restoration of human CD8+ TILs in vitro. Consistently, the combination of ABL503 with anti-PD-1 in vivo significantly alleviated tumor growth, and induced enhanced infiltration and activation of CD8+ TILs. CONCLUSIONS: ABL503-a PD-L1 and 4-1BB dual-targeting bispecific antibody-elicits pronounced additive tumor growth inhibition, with increased infiltration and functionality of exhausted CD8+ T cells, which in turn enhances the anti-cancer effects of anti-PD-1 blockade. These promising findings suggest that ABL503 (TJ-L14B) in combination with PD-1 inhibitors will likely further enhance therapeutic benefit in clinical trials.

Tooth-composite bond failure with a universal and an etch-and-rinse adhesive depending on mode and frequency of application.

OBJECTIVES: To evaluate the effect of an additional layer of universal adhesive on the interfacial enamel/dentin-composite gap formation in relation to application mode and aging, via spectral domain optical coherence tomography (SD-OCT) and scanning electron microscopy (SEM). METHODS: In vitro class V cavities in 114 caries-free premolars were restored by applying one or two layers of a universal adhesive (Scotchbond Universal, SBU) in self-etch (se) and etch-and-rinse (er) mode or the reference adhesive OptiBond FL (OFL-er). The restorations were imaged by SD-OCT (six groups, n = 8) and SEM (n = 3) directly after filling (t1), water storage (t2, 24 h), embedding (t3), and thermo-mechanical loading (t4, TCML). The interfacial gaps were quantified using 26 parameters and analyzed using principal component analysis and linear mixed effect models. RESULTS: Gap formation at enamel and dentin was significantly influenced by the adhesive, the application mode and number of layers (p < 0.001). This was due to the influence of the SBU-er mode (p < 1e-05), which showed significantly more gap formation and a greater range of variation with double application when compared to SBU-se and OFL. The fewest interfacial gaps occurred with one or two applications of OFL-er and one layer of SBU-er. SIGNIFICANCE: Adhesive application mode and the number of adhesive layers are relevant factors in the tooth-composite bond failure. Double application worsened the adaptation of SBU to freshly prepared dentin conditioned with phosphoric acid.

Four-Dimensional Visualization of Microscale Dynamics of Membrane Oil Fouling via Synchrotron Radiation Microcomputed Tomography.

Although oil-water separation technology via wettability-controlled membranes has emerged as a promising technology to treat oily wastewater, membrane fouling by faulents such as sludge flocs and colloids, and the consequent clogging of pores, severely degrades the efficiency of filtration systems. One of the main promotors of fouling by faulents is oil fouling, which is also a form of fouling itself. Despite considerable practical and academic interest in the analysis of oil-fouled membranes, direct visualization of the entire process of oil infiltration into hydrophilic membranes is still preliminary owing to (i) the similar optical contrast and physical density between oil and water, (ii) the low penetration depth of imaging methods, and (iii) the lack of 3D segmentation capability. In this study, microcomputed X-ray tomography using tunable synchrotron radiation provided direct high-speed 3D visualization of the microscale dynamics of the oil infiltration of a prewetted hydrophilic filter membrane over time. Direct visualization of the interfacial dynamics of oil infiltration opens a window into the complex liquid (water/oil)-gas-solid interface and thus helps furnish an in-depth understanding of oil fouling in the prewetted membrane.

Clinical and OCT assessment of application modes of a universal adhesive in a 12-month RCT.

OBJECTIVES: To evaluate a universal adhesive clinically using FDI criteria and by optical coherence tomography (OCT). METHODS: In 50 patients, three or four non-carious cervical lesions (NCCL) were restored with composite (Venus® Diamond Flow, Kulzer) using iBond® Universal (iBU, Kulzer) applied in self-etch (iBU-SE, n = 50), selective-enamel-etch (iBU-SEE, n = 29) or etch-and-rinse mode (iBU-ER, n = 50) and the reference OptiBond™ FL (OFL, Kerr, n = 50). Restorations were imaged by SD-OCT. The weighted mean length of interfacial adhesive defects (AD, %) was quantified per restoration immediately after placement (t0), simultaneously with clinical assessment (FDI criteria) after 14 days (t1), 6 (t2) and 12 months (t3). Data were statistically analyzed (McNemar-/Wilcoxon-/Mann-Whitney-U test (α = 0.05), Kaplan-Meier survival curves). RESULTS: After 12 months, cumulative failure rates were lower with iBU-SE (0.0%; p = 0.016), iBU-SEE (0.0%; p = 0.125), and iBU-ER (2.1%; p = 0.070; loss t3) compared to OFL (16.7%; losses t2, t3). Generally, marginal adaptation decreased (pi < 0.001) and marginal staining increased (pi ≤ 0.031), without significant group differences (pi > 0.064). AD increased in all groups (pi < 0.001). At enamel, AD appeared more extended with iBU-SE vs. iBU-SEE (t2-t3; pi ≤ 0.005), iBU-ER (t1-t3; pi ≤ 0.051) and OFL (t0-t3; pi ≤ 0.018). At dentin/cement iBU generally caused fewer defects than OFL (t1-t3; pi ≤ 0.010) and with SE vs. ER (t2-t3; pi = 0.010). CONCLUSIONS: In NCCLs, iBU generally provides a more durable bond than OFL. Recommended mode is SEE. Clinic and OCT provided comparable results. OCT has higher statistical power, shows group differences earlier and specifically for the different hard tooth tissues. CLINICAL SIGNIFICANCE: The universal adhesive iBU was superior against the reference OFL in NCCLs. It can be recommended with SEE. Evaluation of interfacial adhesive defects by OCT seems to allow early prediction of adhesives' clinical performance.

Arrest of root caries with an adjuvant chlorhexidine-fluoride varnish over a 12-months observation period: a QLF-analyzed, placebo-controlled, randomized, clinical trial (RCT).

This study aimed at evaluating the effectiveness of an adjuvant chlorhexidine-fluoride varnish (Cervitec F) for prevention and arrest of root caries on elderly participants using quantitative light-induced fluorescence (QLF). 23 participants with two or three non-cavitated root carious lesions were included and assigned to three groups of different varnishes (CF: Cervitec F, P: placebo, DP: Duraphate). Agents were applied once to root surface at baseline and in follow-up after 3, 6 and 9 months. The lesions were assessed clinically and with QLF. QLF-images were analyzed regarding fluorescence loss (ΔF), lesion volume (ΔQ) and bacterial activity (ΔR) before (t0), after 14 days (t1), 6- (t2) and 12-months (t3). CF showed a significant difference between t0 and t3: ∆F (- 12.51 [15.41] vs. - 7.80 [16.72], p = 0.012), ∆Q (- 2339.97 (20,898.30) vs. - 751.82 (5725.35), p < 0.001), ∆R (23.80 [41.70] vs. 7.07 [37.50], p = 0.006). Independently of the varnish application, preventive care seems positively influence the root caries progress. Although within CF group the strongest effect was observed, no superiority of a specific varnish application was confirmed over a 12-months QLF observation period. Extra topical fluoride can help remineralise dentin lesions and QLF can be used as a measurement method to determine changes in the dentin lesions.

Evaluation of quantitative light-induced fluorescence to assess lesion depth in cavitated and non-cavitated root caries lesions - An in vitro study.

BACKGROUND: This in vitro study aimed to assess carious lesions on root surfaces using quantitative light-induced fluorescence (QLF) and to compare the readings with axial lesion depth on µCT. METHODS: The root surfaces of 107 extracted human teeth were included after visual-tactile inspection. For further analysis, the following parameters were assessed: clinical findings (non-cavitated: leathery or hard, cavitated), QLF- (QLF-D Biluminator 2+), and µCT-images (Bruker Skyscan 1172). The shape of the undamaged tooth surface of the cavitated lesions was virtually re-constructed during µCT analysis. Clinical surface texture,% fluorescence loss, and lesion depth (µCT) were determined. STATISTICAL ANALYSIS: chi²-test, Spearman-Rho test, regression analysis. RESULTS: ∆F was significantly lower in non-cavitated leathery (-50.37 ± 15.10) and cavitated (-61.23 ± 9.92) compared to non-cavitated surfaces with a hard texture (-17.04 ± 16.10, p < 0.01). For non-cavitated surfaces, a negative correlation was observed between ∆F and lesion depth in µCT images regardless of texture (-0.748, p < 0.01). Regression analysis revealed that ∆F predicted lesion depth in µCT for non-cavitated surfaces (ß: 0.703, CI95: 0.67--0.43, p < 0.01). CONCLUSION: The percentage of fluorescence loss (∆F) in QLF predicted lesion depth of non-cavitated demineralized root surfaces. Therefore, QLF can be recommended for estimating the lesion depth of carious root lesions and seems to expand the possibilities of follow-up and lesion monitoring, especially for non-cavitated surfaces.

Fabrication of Oblique Submicron-Scale Structures Using Synchrotron Hard X-ray Lithography.

Oblique submicron-scale structures are used in various aspects of research, such as the directional characteristics of dry adhesives and wettability. Although deposition, etching, and lithography techniques are applied to fabricate oblique submicron-scale structures, these approaches have the problem of the controllability or throughput of the structures. Here, we propose a simple X-ray-lithography method, which can control the oblique angle of submicron-scale structures with areas on the centimeter scale. An X-ray mask was fabricated by gold film deposition on slanted structures. Using this mask, oblique ZEP520A photoresist structures with slopes of 20° and 10° and widths of 510 nm and 345 nm were fabricated by oblique X-ray exposure, and the possibility of polydimethylsiloxane (PDMS) molding was also confirmed. In addition, through double exposure with submicron- and micron-scale X-ray masks, dotted-line patterns were produced as an example of multiscale patterning.

Light-based diagnostic methods for the in vivo assessment of initial caries lesions: Laser fluorescence, QLF and OCT.

OBJECTIVES: To evaluate different light-based caries diagnostic methods for assessment of non-cavitated initial carious lesions on smooth surfaces. METHODS: 39 participants were included. For each participant, three regions of interest (ROI) with ICDAS codes 0, 1 or 2 were defined. All ROIs (n = 117) were investigated/imaged with laser fluorescence (LF, DIAGNOdent 2095; KaVo dental GmbH), quantitative light-induced fluorescence (QLF; Biluminator 2+, Inspektor Research Systems B.V.) and spectral-domain optical coherence tomography (SD-OCT; Telesto II, Thorlabs GmbH). The values of LF, ΔF of QLF, and the lesion extent assessed by OCT were categorized. Frequency distribution of LF-/QLF- and OCT-scores was determined for each ICDAS code included. Inter- and intra-examiner reproducibility of QLF and OCT measurements were assessed by unweighted kappa coefficient (ĸ) and Wilcoxon test (α = 0.05). RESULTS: Compared with LF and QLF, OCT showed various manifestations of carious lesions for visually sound ROIs and a larger variation of caries extent in depth within the same ICDAS code groups. Intra-examiner reproducibility ranged between 0.49 and 1.00 for the QLF analysis and between 0.95 and 0.99 for the OCT analysis. Inter-examiner reproducibility ranged between 0.17 and 0.32 for the QLF analysis and between 0.65 and 0.79 for the OCT analysis. CONCLUSION: LF and QLF were insensitive to less pronounced smooth-surface lesions while OCT allowed differentiation based on the penetration depth of the carious lesions. This makes OCT a suitable method to complement conventional visual inspection in order to detect and assess (very) early lesions.

Evaluation of low-viscosity bulk-fill composites regarding marginal and internal adaptation.

This study aimed at evaluating the marginal and internal adaptation of low-viscosity bulk-fill composites to enamel and dentin using a self-etch or an etch-and-rinse adhesive without and with artificial ageing. Hundred and twenty-eight MOD cavities in extracted molars were assigned to eight groups (n = 16), restored with the adhesives OptiBond FL (OFL) or Xeno V+ (X) and two low-viscosity bulk-fill composites SDR or x-tra base, covered with Premise. Tetric EvoCeram Bulk Fill and Premise served as a control. n = 8 per group were subjected to prolonged water storage (180 days) and thermocycling (2500×). Scanning electron microscopy was used to determine marginal gaps (MG) and interfacial adhesive defects (IAD). There were no significant differences between composite types in 44 out of 48 (MG) or 43/48 (IAD) comparisons. More MG were observed with X than with OFL (14 out of 16 comparisons, two significant), while in 16 of 16 comparisons with X more IAD were observed (14 significant). After artificial ageing, MG generally increased (9/16 significant), compared to IAD (one significant). The performance of the investigated composite types concerning the integrity of the tooth-composites interface was comparable. Compared to the 1-step self-etch system, the bond with the 3-step etch-and-rinse adhesive was raised.

Direct Visualization of Microscale Dynamics of Water Droplets on under-Oil-Hydrophilic Membranes by Using Synchrotron White-Beam X-ray Microimaging Techniques.

Despite considerable academical and practical interests on separation of water-in-oil emulsion via special wettable membranes, fundamental understanding on microscale dynamics of water droplets on under-oil-hydrophilic membranes (UOHMs) at early stages during separation is still very preliminary due to temporal and spatial resolution of existing visualization techniques. To this end, we here succeed in a direct microscopic visualization of separation processes of water droplets on the UOHMs by employing a high-speed, two-dimensional synchrotron white-beam X-ray microimaging technique. During the separation of water-in-oil emulsion, microscale dynamic behaviors of water droplets on hydrophilic membrane surfaces immersed in the different oil media (i.e., hexane, kerosene, and light and heavy mineral oils) and oil films between water droplets and membrane surfaces are visualized and analyzed.

Assessment of non-cavitated root caries lesions by quantitative light-induced fluorescence-An in vivo feasibility study.

OBJECTIVES: To evaluate the reproducibility of quantitative light-induced fluorescence (QLF) for the detection and assessment of demineralization states of non-cavitated root surfaces and to determine the correlation between visual inspection and QLF-analysis. METHODS: 46 exposed non-cavitated root surfaces of 12 participants were classified in three scores: sound (0), lesion ≤ 5 mm (1a), lesion > 5 mm in diameter (1b). Three examiners imaged every root surface three times using QLF (QRayCam) and measured fluorescence loss (ΔF) and lesion volume (ΔQ). The QLF-images were analyzed by three examiners. The intra- and interexaminer reproducibilities were calculated (intraclass correlation coefficient, ICC). The correlation between the scores of root caries and QLF-analysis was determined (rank correlation coefficient, Spearman-Rho ρ). RESULTS: Intra- and interexaminer reproducibility (ICC) was 0.98 and 0.95 for ΔF, 0.94 and 0.91 for ΔQ, respectively. A significant correlation was observed between the scores and ΔF (ρ = -0.53, p < 0.01). CONCLUSION: QLF as a reproducible diagnostic tool enables non-invasive detection and differentiation of non-cavitated root caries lesions on root surfaces in vivo. It can be recommended in addition to the clinical evaluation for prospective follow-up examinations of demineralization states of exposed root surfaces.

Clinical and OCT outcomes of a universal adhesive in a randomized clinical trial after 12 months.

OBJECTIVES: To assess the performance of a universal adhesive in different application modes in non-carious cervical lesions clinically and by optical coherence tomography (OCT). METHODS: 55 adult patients with three non-carious cervical lesions (NCCL) each participated in the study. Lesions were restored with Scotchbond™ Universal (SBU, 3 M) applied in the self-etch (SBU-SE) and the selective-enamel-etch mode (SBU-SEE) in combination with Filtek™ Supreme XTE (3 M). OptiBond™ FL (OFL, Kerr) was used as a control. Restorations were clinically assessed (FDI criteria) after 14 days, 6 and 12 months and in parallel imaged by OCT (interfacial adhesive defects), starting immediately after filling placement. Cumulative failure rates (CFR) and means of interfacial adhesive defect were statistically evaluated. RESULTS: After 12 months, CFRs were lower in the SBU groups (0.0% each) than in the OFL group (20.0%, p = 0.001). Clinically, small marginal fractures occurred three times more often in the SBU-SE than in the SBU-SEE group (p = 0.001). Immediately after filling placement and at each reassessment OCT revealed more interfacial defects at enamel interfaces for SBU/SE compared to SBU/SEE and OFL (pi ≤ 0.044). At dentin/cement more defects were seen with OFL compared to SBU/SE and SBU/SEE (pi ≤ 0.001). Before restoration loss, more interfacial defects appeared compared to remaining restorations (pimmediately/6M = 0.132/0.002). CONCLUSIONS: Clinical evaluation and OCT imaging revealed higher interfacial integrity for SBU in both application modes compared to OFL. OCT detected interfacial bond failures prior to clinical deterioration or restoration loss. CLINICAL SIGNIFICANCE: Scotchbond Universal showed an equivalent or improved bonding performance compared to the reference adhesive. Selective enamel etching is recommended. The parameter interfacial adhesive defect seems to be a valuable predictor for evaluation of adhesive restoration systems.

Effectiveness of caries-preventing agents on initial carious lesions within the scope of orthodontic therapy.

OBJECTIVE: To evaluate the effectiveness of three different caries-preventing agents on artificial caries in a Streptococcus mutans-based caries model. METHODS: Sixty-five caries-free human molar enamel blocks were treated with a demineralization solution and a remineralization solution. The specimens were assigned to the following groups according to the caries-protective product applied: group A, chlorhexidine varnish; group B, fluoride-releasing chemically cured sealant; group C, fluoride-releasing lightcured sealant; group D, positive control (specimens that were subjected to de- and remineralization cycles without treatment with any caries-protective agents); and group E, negative control (specimens that were not subjected to de- and remineralization cycles). Samples in groups A-D were stored in demineralization solution with S. mutans and thereafter in artificial saliva. This procedure was performed for 30 days. Average fluorescence loss (ΔF) and surface size of the lesions were measured using quantitative light-induced fluorescence at baseline and on the 7th, 14th, and 30th days. RESULTS: After 30 days, group A demonstrated a significant increase in ΔF and the surface size of the lesions, no significant difference in comparison with the positive control group, and a significant difference in comparison with the negative control group. Group B showed no significant changes in both parameters at any of the measurement points. While group C showed increased ΔF after 14 days, no significant fluorescence change was observed after 30 days. CONCLUSIONS: Both fluoride-releasing sealants (chemically or light-cured) show anti-cariogenic effects, but the use of chlorhexidine varnish for the purpose of caries protection needs to be reconsidered.

Correction to: Internal and marginal adaptation of high-viscosity bulk-fill composites in class II cavities placed with different adhesive strategies.

In the original publication of the article the paragraph "The four resin-based composites…" appearing below the "Materials and methods" section is incomplete. The correct paragraph is provided in this correction below.

Internal and marginal adaptation of high-viscosity bulk-fill composites in class II cavities placed with different adhesive strategies.

The aim of this study was to evaluate internal and marginal adaptation of high-viscosity bulk-fill composites to enamel and dentin with a self-etch (SE) and an etch-and-rinse adhesive (ER) without and with artificial aging. 128 MOD cavities in extracted human molars were prepared, randomly assigned to 8 groups (n = 16), bonded with either OptiBond FL (OFL, ER) or Xeno V+ (X, SE), and restored in 4 mm horizontal bulk layers with SonicFill (SF), Tetric EvoCeram Bulk fill (TEC), and x-tra fil (XF). Incremental layering technique with Premise (P) served as control. Half of the specimens each (n = 8) were subjected either to water storage (1 day, 37 °C) or prolonged storage (180 days) and thereafter thermocycling. Replicas were analyzed for marginal gap formation. After sectioning, interfacial adhesive defects were assessed. Results were statistical analyzed. (1) Internal adaptation: Except for TEC/OFL at enamel without artificial aging, no significant differences between bulk-fill composites and the control were observed. All groups at dentin with OFL showed less internal adhesive defects than that with X (p < 0.05). (2) Marginal adaptation: No significant differences were observed between bulk-fill composites and control except for P after artificial aging (p > 0.05). All other composites, regardless of artificial aging, formed significantly more marginal gaps at enamel with X compared to with OFL (p ≤ 0.05). Simplified restorations with high-viscosity bulk-fill composite showed comparable internal and marginal adaptation to incrementally placed fillings. A superiority of the 3-step ER approach was seen compared to the 1-step SE adhesive irrespective of the bulk-fill composite used.

Fabrication of Polymer Microstructures of Various Angles via Synchrotron X-Ray Lithography Using Simple Dimensional Transformation.

In this paper, we developed a method of fabricating polymer microstructures at various angles on a single substrate via synchrotron X-ray lithography coupled with simple dimensional transformations. Earlier efforts to create various three-dimensional (3D) features on flat substrates focused on the exposure technology, material properties, and light sources. A few research groups have sought to create microstructures on curved substrates. We created tilted microstructures of various angles by simply deforming the substrate from 3D to two-dimensional (2D). The microstructural inclination angles changed depending on the angles of the support at particular positions. We used convex, concave, and S-shaped supports to fabricate microstructures with high aspect ratios (1:11) and high inclination angles (to 79°). The method is simple and can be extended to various 3D microstructural applications; for example, the fabrication of microarrays for optical components, and tilted micro/nanochannels for biological applications.

OCT for early quality evaluation of tooth-composite bond in clinical trials.

OBJECTIVES: To evaluate early quality of composite restorations with a universal adhesive in different application modes clinically and with optical coherence tomography (OCT). METHODS: 22 patients with four non-carious cervical lesions each received composite restorations (Filtek Supreme™ XTE, 3 M). The universal adhesive Scotchbond Universal™(SBU, 3 M) was applied with three etching protocols: self-etch (SE), selective-enamel-etch (SEE) and etch-and-rinse (ER). The etch-and-rinse adhesive OptiBond™ FL (OFL, Kerr) served as a control. Restorations were imaged by OCT (Thorlabs) directly after application (t0). After 14 days (t1) and 6 month (t2) OCT imaging (interfacial adhesive defects) was repeated combined with clinical assessment (FDI criteria). Groups were compared by Friedman-/Wilcoxon- and McNemar-Test. RESULTS: No differences were seen clinically between groups (pi ≥ 0.500). OCT assessment revealed more adhesive defects at the enamel interface with SBU/SE at t0-t2 compared to all groups (pi ≤ 0.016). OFL showed more defects than SBU/ER (t1: p = 0.01; t2: p = 0.083). At dentin/cementum interface OFL exhibited more adhesive defects than SBU with all conditioning modes (t0, t1, pi ≤ 0.003) and at t2 to SBU/SE and SBU/ER (p < 0.001). Since t1 defects with SBU were detected more frequently in the SE and SEE modes compared to ER (pi ≤ 0.037). In contrast to SBU defects increased with OFL up to t2 (pi ≤ 0.007). CONCLUSIONS: In contrast to clinical evaluation, OCT revealed subtle adhesive defects directly after application that might interfere with clinical success. It was demonstrated that ER does not decrease initial adhesion of SBU to dentin.

Optical coherence tomography to evaluate variance in the extent of carious lesions in depth.

Evaluation of variance in the extent of carious lesions in depth at smooth surfaces within the same ICDAS code group using optical coherence tomography (OCT) in vitro and in vivo. (1) Verification/validation of OCT to assess non-cavitated caries: 13 human molars with ICDAS code 2 at smooth surfaces were imaged using OCT and light microscopy. Regions of interest (ROI) were categorized according to the depth of carious lesions. Agreement between histology and OCT was determined by unweighted Cohen's Kappa and Wilcoxon test. (2) Assessment of 133 smooth surfaces using ICDAS and OCT in vitro, 49 surfaces in vivo. ROI were categorized according to the caries extent (ICDAS: codes 0-4, OCT: scoring based on lesion depth). A frequency distribution of the OCT scores for each ICDAS code was determined. (1) Histology and OCT agreed moderately (κ = 0.54, p ≤ 0.001) with no significant difference between both methods (p = 0.25). The lesions (76.9% (10 of 13)) _were equally scored. (2) In vitro, OCT revealed caries in 42% of ROI clinically assessed as sound. OCT detected dentin-caries in 40% of ROIs visually assessed as enamel-caries. In vivo, large differences between ICDAS and OCT were observed. Carious lesions of ICDAS codes 1 and 2 vary largely in their extent in depth.

Handheld Photoacoustic Microscopy Probe.

Optical resolution photoacoustic microscopy (OR-PAM) is a non-invasive, label-free method of in vivo imaging with microscopic resolution and high optical contrast. Based on intrinsic contrasts, OR-PAM has expanded to include in vivo vessel imaging, flow cytometry, physiological parameter analysis, and single-cell characterization. However, since conventional OR-PAM systems have a fixed tabletop configuration, a large system size, and slow imaging speed, their use in preclinical and clinical studies remains limited. In this study, using microelectromechanical systems (MEMS) technology, we developed a handheld PAM probe with a high signal-to-noise ratio and image rate. To enable broader application of the OR-PAM system, we reduced its size and combined its fast scanning capabilities into a small handheld probe that uses a 2-axis waterproof MEMS scanner (2A-WP-MEMS scanner). All acoustical, optical, and mechanical components are integrated into a single probe with a diameter of 17 mm and a weight of 162 g. This study shows phantom and in vivo images of various samples acquired with the probe, including carbon fibers, electrospun microfibers, and the ear, iris, and brain of a living mouse. In particular, this study investigated the possibility of clinical applications for melanoma diagnosis by imaging the boundaries and morphology of a human mole.