A new laser fluorescence method, KaVo DIAGNOdent, was tested with respect to reproducibility and validity and compared with radiography regarding accuracy in the detection of occlusal caries. Seventy–six extracted premolar and molar teeth were measured twice with DIAGNOdent under both wet and dry conditions, at an interval of 2 weeks. Conventional film radiographs were exposed. Microradiographic analysis of sections of the teeth served as the gold standard. Intra–class correlation coefficient and Pearson’s correlation coefficient were used to assess the reproducibility and the validity of the method, respectively. The influence of time and varying humidities on DIAGNOdent readings were analysed by two–way repeated measure ANOVA. ROC curves were plotted for DIAGNOdent readings and radiographic registration of caries by 6 observers, and the areas under the curves were compared using Student’s t test. Under both wet and dry conditions, the reproducibility of the DIAGNOdent method was excellent: R = 0.97 and 0.96, respectively. The diagnostic accuracy of DIAGNOdent was significantly better than that of radiography (p≤0.001). In this in vitro study of detection of occlusal caries, the diagnostic performance of the DIAGNOdent method was superior to that of radiography.

The aims were 1) to validate a laser-based device, KaVo DIAGNOdent, for quantification of caries lesions on smooth surfaces, using histopathologic and microradiographic analyses as the gold standard, and 2) to test inter- and intra-observer agreements. We also investigated the influence on DIAGNOdent readings of the storage medium used for extracted teeth. Two observers measured independently the tooth surfaces of 40 extracted premolars that had been stored in thymol-saturated saline. After subsequent storage in neutral-buffered formalin for 14 days they were re-measured. The teeth were then sectioned for histopathologic and microradiographic analysis. The Spearman rank correlation coefficients between lesion depth and DIAGNOdent readings were 0.78-0.83 and 0.85 for teeth stored in thymol-saturated saline and formalin, respectively. Inter- and intra-observer agreements were 0.94 and 0.95 when the teeth were stored in thymol-saturated saline. The DIAGNOdent reading was almost 1.5 times higher for teeth stored in formalin than for those stored in thymol saline. The Spearman rank correlation coefficient between mineral loss in enamel (deltaZ) and DIAGNOdent readings ranged from 0.64 to 0.68. It was concluded that DIAGNOdent may be helpful for assessing smooth-surface caries, but the cut-off points need to be assessed under clinical conditions. The increase in fluorescence associated with storage of teeth in formalin warrants further investigation.

BackgroundAutologous grafting, despite some disadvantages, is still considered the gold standard for reconstruction of maxillofacial bone defects. The aim of this study was to evaluate bone regeneration using bone marrow-derived mesenchymal stromal cells (MSCs) in a clinical trial, a less invasive approach than autologous bone grafting. This comprehensive clinical trial included subjects with severe mandibular ridge resorption.MethodsThe study included 11 subjects aged 52–79 years with severe mandibular ridge resorption. Bone marrow cells were aspirated from the posterior iliac crest and plastic adherent cells were expanded in culture medium containing human platelet lysate. The MSCs and biphasic calcium phosphate granules as scaffolds were inserted subperiosteally onto the resorbed alveolar ridge. After 4–6 months of healing, new bone formation was assessed clinically and radiographically, as were safety and feasibility. Bone at the implant site was biopsied for micro-computed topography and histological analyses and dental implants were placed in the newly regenerated bone. Functional outcomes and patient satisfaction were assessed after 12 months.ResultsThe bone marrow cells, expanded in vitro and inserted into the defect together with biphasic calcium phosphate granules, induced significant new bone formation. The regenerated bone volume was adequate for dental implant installation. Healing was uneventful, without adverse events. The patients were satisfied with the esthetic and functional outcomes. No side effects were observed.ConclusionsThe results of this comprehensive clinical trial in human subjects confirm that MSCs can successfully induce significant formation of new bone, with no untoward sequelae. Hence, this novel augmentation procedure warrants further investigation and may form the basis of a valid treatment protocol, challenging the current gold standard.Trial registrationEudraCT, 2012-003139-50. Registered on 21 August 2013. ClinicalTrials.gov, NCT 02751125. Registered on 26 April 2016.

It is hypothesized that occlusal lesions are initiated on the fissure walls and can therefore be obscured by sound superficial tissue. Additionally, there is evidence that one effect of regular use of fluorides is greater opacity of enamel, which may obscure underlying lesions in dentin, the so-called 'hidden lesions'. Dental radiographs are inadequate for detecting decay in the occlusal surfaces until the lesion is well advanced through the enamel and into the dentin. The clinician relies on visual observation of texture and discoloration, clinical judgment based upon experience, and on tactile sense by probing with an explorer. An objective detection method to complement the traditional visual assessment is used by the clinician for arriving at clinical decisions on the management of the carious lesion: whether invasive therapy or a more conservative, noninvasive approach. Objective and reliable longitudinal monitoring of the lesion's response to preventive measures allow the selection of an appropriate therapy before the lesion progresses to the stage where invasive treatment is required. This paper discusses the problem of the lack of appropriate clinical methods for the detection and quantification of carious lesions. A few commercially available methods are described (the quantitative light-induced fluorescence method, the DIAGNOdent device, and electrical caries monitor) and some new techniques mentioned.

In vivo repeatability and reproducibility of the quantitative light-induced fluorescence (QLF) method were tested with respect to three variables: lesion area, and average and maximum changes in lesion fluorescence. To test the image-capturing stages, three analysts each captured images of 15 incipient smooth surface lesions in vivo, and the images were analysed by one of the analysts. To test the analytical stage of the method, three analysts analysed the images of 15 in vivo incipient smooth surface lesions. For the image-capturing stage, inter-examiner reliability showed an intra-class correlation coefficient (r) between 0.95 and 0.98. For the analytical stage, intra-examiner reliability for all three analysts showed a value of r between 0.93 and 0.99. Inter-examiner reliability showed a value of r between 0.95 and 0.99. It was concluded that the in vivo repeatability and reproducibility of the QLF method are excellent.

Ecdysone 20-monooxygenase (E20MO), a cytochrome P450 monooxygenase (CYP314A1), catalyses the conversion of ecdysone (E) to 20hydroxyecdysone (20E). We report here the cloning and characterization of the Halloween gene Shade (Shd) encoding E20MO in the Colorado potato beetle, Leptinotarsa decemlineata. LdSHD has five conserved motifs typical of insect P450s, ie the Helix-C, Helix-I, Helix-K, PxxFxPE/DRF (PERF) and heme-binding motifs. LdShd was expressed in developing eggs, the first to fourth instars, wandering larvae, pupae and adults, with statistically significant fluctuations. Its mRNA was ubiquitously distributed in the head, thorax and abdomen. The recombinant LdSHD protein expressed in Spodoptera frugiperda 9 (Sf9) cells catalysed the conversion of E to 20E. Dietary introduction of double-stranded RNA (dsRNA) of LdShd into the second instar larvae successfully knocked down the LdShd expression level, decreased the mRNA level of the ecdysone receptor (LdEcR) gene, caused larval lethality, delayed development and affected pupation. Moreover, ingestion of LdShd-dsRNA by the fourth instars also down-regulated LdShd and LdEcR expression, reduced the 20E titre, and negatively influenced pupation. Introduction of 20E and a nonsteroidal ecdysteroid agonist halofenozide into the LdShd-dsRNA-ingested second instars, and of halofenozide into the LdShd-dsRNA-ingested fourth instars almost completely relieved the negative effects on larval performance. Thus, LdSHD functions to regulate metamorphotic processes by converting E to 20E in a coleopteran insect species Le. decemlineata.

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DIAGNOdent is now well documented for detection of primary occlusal caries, but not as yet for secondary caries. The aim of this study was to investigate the application of DIAGNOdent for in vitro detection of secondary caries. The material comprised 66 extracted teeth: 48 with amalgam restorations and 18 with tooth-colored restorations. Digital images of occlusal surfaces were captured for each specimen. The teeth were mounted in groups of 3 or 4 in plaster blocks simulating their anatomical positions. Bitewing radiographs of each block were assessed for secondary caries by 5 observers. The margins of each restoration were carefully scanned with DIAGNOdent; the site of the highest reading was localized in digital images; and the corresponding values were registered. Marginal integrity and staining were also documented. The restorations were then removed and the teeth were hemi-sectioned. For verification, two observers working together examined all the cavities under a stereomicroscope at 16 x magnification followed by probing. Sensitivity and specificity for DIAGNOdent and conventional radiography were 0.77/0.81 and 0.65/0.81, respectively. Regarding ROC analyses, the Az values were 0.89 and 0.72 for DIAGNOdent and radiography. For DIAGNOdent, the false-positive fraction included only stained teeth. Cohen's kappa statistics disclosed moderate agreement between the 2 methods, with an agreement of 56%. DIAGNOdent tended to give more positive diagnoses than bitewing radiography. The results indicate that DIAGNOdent may be a helpful tool for detecting secondary caries.