Search Results (16)

Background/Objectives: Cutaneous squamous cell carcinoma (cSCC) is the second most common skin cancer, with diverse clinical presentations. This study aims to correlate findings from dermoscopy, ultrasonography, ex vivo confocal microscopy, and histology to improve diagnostic accuracy and guide better clinical management of cSCC. Methods: This cross-sectional study, conducted between July 2022 and December 2024, included 26 patients with 35 clinically suspicious cSCC tumors, analyzed through clinical, dermoscopic, high-frequency ultrasound (HFUS), ex vivo confocal fluorescence microscopy (FCM), and histopathology. Tumors were evaluated for various clinical, imaging, and histopathological criteria, such as tumor thickness, vascularization, differentiation degree, and invasion level, with FCM applied to 24 tumors for advanced microscopic analysis. Results: The study analyzed 35 cases of histopathologically confirmed cSCC, finding that invasive SCC was associated with greater tumor thickness, increased vascularization, and ulceration on both ultrasound and dermatoscopy, while in situ SCC showed homogeneous echogenicity and specific dermoscopic patterns like dotted vessels and white halos. Strong correlations were identified between ultrasound and histopathological measurements of tumor thickness and invasion depth, and confocal microscopy revealed that features like plump bright cells and nest-like structures were linked to invasive and poorly differentiated tumors. Conclusions: This study uniquely integrates advanced imaging techniques—dermatoscopy, skin ultrasound, and ex vivo confocal microscopy—with histopathological analysis to provide new insights into tumor grade, vascularity, and invasion depth in cSCC, enhancing non-invasive diagnosis. Full article

The impact of adding silver nanoparticles (AgNPs) to bioceramic (BC) sealer on their ability to penetrate dentinal tubules is still unknown. Thus, this confocal laser scanning microscopic (CLSM) study aimed to assess the extent of dentinal tubule penetration of BC sealer (TotalFill^® Hiflow BC Sealer™, FKG, Switzerland) with and without AgNPs using the single-cone (SC) technique and the continuous-wave condensation (CWC) technique. AgNPs alone as well as in a mixture with the BC sealer were characterized using scanning electron microscopy and transmission electron microscopy. Single-rooted extracted human teeth (N = 100) were selected and prepared, and then divided into four groups (n = 25). Group 1 (BC/SC): BC sealer obturated with the SC technique. Group 2 (BC+AgNPs/SC): BC sealer with AgNPs obturated with the SC technique. Group 3 (BC/CWC): BC Sealer obturated with the CWC technique. Group 4 (BC+AgNPs/CWC): BC Sealer with AgNPs obturated with the CWC technique. After 2 weeks, roots were horizontally sectioned to obtain 1 mm thick dentin slices that were evaluated with CLSM. Sealer dentinal tubule penetration area and the maximum depth of penetration were measured. Data were analyzed with one-way ANOVA and the Tukey multiple comparison tests (p ≤ 0.05). The characterization process demonstrated a spherical-shaped nanoparticles without obvious agglomeration. The results showed that Group 2 (BC+AgNPs/SC) significantly demonstrated the highest mean tubular penetration depth, while group 3 (BC/CWC) had the lowest mean depth. Group 2 (BC+AgNPs/SC) exhibited the significantly highest mean value for the total area of penetration. However, groups 1 (BC/SC) and 3 (BC/CWC) exhibited the lowest mean value of total penetration area, with no statistically significant difference. The integration of AgNPs with BC sealer markedly enhanced penetration into dentinal tubules. The SC technique demonstrated superior penetration relative to the CWC technique. Full article

Multiple sclerosis (MS) is a chronic neurodegenerative disorder involving demyelination. The cuprizone model is commonly used to study MS by inducing oligodendrocyte stress and demyelination. The subventricular zone (SVZ) plays a key role in neurogenesis, while the neuronal/glial antigen 2 (NG2) is a marker for immature glial cells, involved in oligodendrocyte differentiation. The apelin receptor (APLNR) is linked to neurogenesis and behavior modulation. This study explores the role of APLNR in NG2-positive cells during de- and remyelination phases in the experimental cuprizone mouse model. Thirty male C57BL/6 mice were divided into control (not treated), demyelination (5 weeks cuprizone administration), and remyelination (5 weeks cuprizone administration + 5 weeks recovery) groups. Histological examinations, immunohistochemistry, and immunofluorescence on serial coronal sections were conducted to evaluate corpus callosum (CC) morphology and APLNR and NG2 expression in the SVZ, in addition to behavioral assessments. The histological analysis showed a significant reduction in the CC’s thickness and area after five weeks of cuprizone exposure, followed by recovery five weeks post-exposure. During the demyelination phase, APLNR-expressing cells peaked while NG2-positive cells decreased. In the remyelination phase, APLNR-expressing cells declined, and NG2-positive cells increased. Confocal microscopy confirmed the co-localization of NG2 and APLNR markers. Statistically significant differences were observed across experimental groups. Correlation analyses highlighted associations between APLNR/NG2 cell counts and CC changes. Behavioral tests revealed impaired motor coordination and memory during demyelination, with gradual recovery during remyelination. Significant changes in the CC structure and the number of APLNR and NG2-positive cells were observed during de- and remyelination, suggesting that NG2-positive cells expressing APLNR may play a key role in remyelination. Full article

In this work, ethyl cellulose was used as a wall material, propanetriol as a core material, polyvinyl alcohol as a stabilizer and gelatin as an emulsifier. Self-healing microcapsules with a slow-release effect were prepared using the solvent evaporation method. Various analytical techniques, such as 3D confocal microscopy (LCSM), optical microscopy (OM), scanning electron microscopy (SEM), infrared spectroscopy (FT-IR), energy dispersive spectroscopy (EDS), thermal weight loss analysis (TGA), laser particle size tester and electrochemical impedance polarization, are utilized. The morphology, distribution, particle size, corrosion resistance and self-healing ability of the prepared microcapsules and resin-based coatings were characterized and analyzed. The results show that the cross-sectional core–shell structure is clearly seen in the LCSM, showing a smooth, hollow, spherical shape. OM and laser particle size testers have shown that the size of the microcapsules decreases over time. Also, in OM, the microcapsules are uniformly distributed in the emulsion with a smooth and non-adherent surface. In SEM, the microcapsule particle size is about 150 μm, the shell wall thickness is about 18 μm, and the hollow structure of ruptured microcapsules is obvious. FT-IR and TGA confirmed the successful encapsulation of the formulated microcapsules. The results show that when the core-wall mass ratio is 1.2:1 and the amount of microcapsule is 10% of the coating amount, the prepared microcapsule has high thermal stability and certain wear resistance. By electrochemical and immersion experiments, it was found that a 3.5 wt % NaCl solution has the best impedance, the lowest corrosion current density, and good adhesion and tensile toughness. The results showed that glycerol was successfully released from the broken microcapsules and self-healed, forming an anticorrosive coating with excellent corrosion resistance and self-healing ability. Full article

Background: The proper sealing of the root canal seems to be critical to obtain a stable result of endodontic therapy. The sealer’s penetration into dentinal tubules during root canal treatment is probably a crucial factor to provide better clinical results. The aim of this study was to compare the effect of three irrigation protocols on the penetration of the epoxy sealer into dentinal tubules and two dyes used to stain the sealer. Methods: Ninety single-canaled human teeth with straight roots were used. The root canals were instrumented with Reciproc instruments up to the size 40/04 ISO. The teeth were divided into three groups (n = 30). The root canals of each group were rinsed accordingly: the control group 1: 5.25% sodium hypochlorite (NaOCl); the experimental group 2: smear layer removal (two times alternatively: 40% citric acid and 5.25% NaOCl) and NaOCl; the experimental group 3: smear layer removal (same as in group 2) and isopropyl alcohol. In each group the roots were further divided into 2 subgroups (n = 15). The root canals were obturated using warm vertical compaction technique of gutta-percha with AH Plus sealer marked with hydrophilic fluorescein (subgroup F) or hydrophobic porphyrin (subgroup P). After 72 h, one-millimeter-thick cross-sections were cut in two, five and eight millimeters distance from the apex. The depth of the penetration of the sealer into the dentinal tubules (resin tags) was measured with the use of a confocal laser microscope. Results: In of all the root parts, the longest resin tags were observed in group 2, whereas the shortest ones were found in group 1 (in the porphyrin subgroups all differences were statistically significant). Within the fluorescein subgroups, the differences between all groups were statistically significant in the middle section of the roots. In the apical and the coronal sections, significant differences were observed between group 1 and the other two groups. Conclusions: The isopropyl alcohol at the end of the irrigating protocol did not affect the higher sealer penetration of the sealer into the dentinal tubules compared with sodium hypochlorite. With the limitation of this study, the hydrophobic porphyrin may be considered as the favorable dye choice to stain endodontic sealers in further studies with confocal laser scanning microscopy, but the methods and reagents used should still be in the research phase. Full article

Background: Once the chemo-mechanical preparation of root canals is finished, achieving a complete seal of the root canal system becomes crucial in determining the long-term success of endodontic treatment. The important goals of root canal obturation are to minimize leakage and achieve an adequate seal. Thus, a material that possesses satisfactory mechanical characteristics, is biocompatible, and has the ability to penetrate the dentine tubules adequately is needed. Aim: This study aimed to compare the penetrability and bond strength between two calcium silicate-based sealers and an epoxy resin-based sealer, as well as examine the relationship between penetrability and bond strength for the different sealers. Method and materials: Thirty-nine recently extracted single-rooted human premolar teeth were instrumented and divided evenly into three groups (n = 13), according to the sealer used for obturation: AH Plus Jet, EndoSequence, and AH Plus Bioceramic Sealer. Three teeth (30 slices) were randomly selected out of each for analysis using confocal laser scanning microscopy to assess penetrability. The remaining ten teeth (90 slices) in each group were subject to push-out tests using a universal testing machine. All teeth were sectioned into nine transverse slices of 0.9 mm thickness for their respective tests (apical, middle, coronal). Results: AH Plus Jet exhibited significantly lower penetrability and significantly higher bond strength compared to EndoSequence BC sealer (p = 0.002) and AH Plus Bioceramic Sealer (p = 0.006). There was no significant difference between EndoSequence BC sealer and AH Plus Bioceramic Sealer in terms of either penetrability or bond strength. No correlation was found between penetrability and bond strength. Conclusions: Within the limitation of this study and regardless of the location in the canal, the bioceramic based root canal sealers appeared to perform better than the epoxy resin-based sealer in terms of dentinal penetration rate. Further studies are required to compare other biomechanical properties of bioceramic sealers including setting characteristics and bacterial leakage. Full article

Objectives: Tissue breakdown was assessed by confocal laser scanning microscopy (CLSM) using autofluorescence around implants with ligatures, on a dog hemimandible. Influence of section thickness on the accuracy of histometrical observations was also evaluated, in comparison with thin sections in light microscopy. Material and Methods: Three months after tooth extraction, implants were placed. Two months after abutment placement, ligatures were placed with no plaque control. 11 months post-implantation, the animal was sacrificed. Undecalcified thin (30 µm) sections were cut, stained and evaluated by light microscopy to be used as a reference. Additional sections were performed, so that another pair of unstained thick sections resulted (250–300 µm). Tissue loss was assessed using histomorphometric parameters under CLSM and was compared to the light microscopy reference ones. Results: Morphometry confirmed tissue loss more pronounced on the “thick” and quick sections, when compared to the time-consuming and technique-sensitive “thin” ones. Conclusions: Within the limits of the present study, the adequacy of histometrical observations under CLSM reveal commensurable information about soft-tissue-bone-implant details, when compared to traditional light microscopy histological protocols. The CLSM investigation may seem demanding, yet the richness of data acquired may justify this approach, provided seatbacks caused by improper manipulation of “thick” sections are avoided. Full article

The Taxodium hybrid Zhongshanshan fast-growing species is susceptible to environment and gravity to form reaction wood. In this study, individual growth rings of reaction wood are used as subjects, and an individual growth ring is divided into three zones: compression zone (CZ), lateral zone (LZ), and opposite zone (OZ). The microanatomical structure and chemical properties of the tracheids in CZ, LZ, and OZ forms by the inclined or bent growth of T. Zhongshanshan are comparatively analyzed by using optical microscopy, scanning electron microscope, laser confocal microscopy, and Raman imaging techniques. In CZ, the length and diameter of compression wood (CW) tracheids decreased, and the shape of cross-sections became rounded as compared to the OZ and LZ tracheids. More notably, threaded fissures appeared on the cell wall of tracheids, and the thickness of the cell wall increased in CW. The analysis of tracheids’ cell wall structure showed that CW tracheids had a complete outer secondary wall middle (S2L) layer, but had no secondary wall inner (S3) layer. In the transition zone (TA) between CW and normal early wood, tracheids were divided into compressed and normal tracheids. Despite the compressed tracheids having a similar cell morphology to normal tracheids, they had a thin secondary wall S2L layer. Tracheids in LZ had a thin S2L layer only at the angle of the cell. No S2L layer was seen in the cell wall of OZ and CZ late wood tracheids. It can be concluded that the response of lignin deposition location to external stress was faster than the change in cell morphology. The above results help provide the theoretical basis for the response mechanism of T. Zhongshanshan reaction wood anatomical structures to the external environment and has important theoretical value for understanding its characteristics and its rational and efficient usage. Full article

Respiratory disease in horses is caused by a multifactorial complex of infectious agents and environmental factors. An important pathogen in horses is equine herpesvirus type 1 (EHV-1). During co-evolution with this ancient alphaherpesvirus, the horse’s respiratory tract has developed multiple antiviral barriers. However, these barriers can become compromised by environmental threats. Pollens and mycotoxins enhance mucosal susceptibility to EHV-1 by interrupting cell junctions, allowing the virus to reach its basolateral receptor. Whether bacterial toxins also play a role in this impairment has not been studied yet. Here, we evaluated the role of α-hemolysin (Hla) and adenylate cyclase (ACT), toxins derived from the facultative pathogenic bacterium Staphylococcus aureus (S. aureus) and the primary pathogen Bordetella bronchiseptica (B. bronchiseptica), respectively. Equine respiratory mucosal explants were cultured at an air–liquid interface and pretreated with these toxins, prior to EHV-1 inoculation. Morphological analysis of hematoxylin–eosin (HE)-stained sections of the explants revealed a decreased epithelial thickness upon treatment with both toxins. Additionally, the Hla toxin induced detachment of epithelial cells and a partial loss of cilia. These morphological changes were correlated with increased EHV-1 replication in the epithelium, as assessed by immunofluorescent stainings and confocal microscopy. In view of these results, we argue that the ACT and Hla toxins increase the susceptibility of the epithelium to EHV-1 by disrupting the epithelial barrier function. In conclusion, this study is the first to report that bacterial exotoxins increase the horse’s sensitivity to EHV-1 infection. Therefore, we propose that horses suffering from infection by S. aureus or B. bronchiseptica may be more susceptible to EHV-1 infection. Full article

In vitro transcribed messenger ribonucleic acid (mRNA) constitutes an emerging therapeutic class with several clinical applications. This study presents a systematic comparison of different technologies—intradermal injection, microneedle injection, jet injection, and fractional laser ablation—for the topical cutaneous delivery of mRNA. Delivery of Cy5 labeled mRNA and non-labeled enhanced green fluorescent protein (eGFP) expressing mRNA was investigated in a viable ex vivo porcine skin model and monitored for 48 h. Forty 10 µm-thick horizontal sections were prepared from each skin sample and Cy5 labeled mRNA or eGFP expression visualized as a function of depth by confocal laser scanning microscopy and immunohistochemistry. A pixel-based method was used to create a semi-quantitative biodistribution profile. Different spatial distributions of Cy5 labeled mRNA and eGFP expression were observed, depending on the delivery modality; localization of eGFP expression pointed to the cells responsible. Delivery efficiencies and knowledge of delivery sites can facilitate development of efficient, targeted mRNA-based therapeutics. Full article

The aim of the present study was to evaluate the influence of several experimental pretreatment crosslinker solutions on the resin polymer–dentine interface created using a representative universal adhesive system, by means of microtensile bond strength testing (μTBS), nanomechanical properties and ultramorphology confocal laser scanning microscopy (CLSM). Five experimental solutions containing different flavonoids were applied as dentine pretreatment after acid etching. A control pretreatment group containing no flavonoid was also employed. A representative modern universal adhesive was then applied, followed by a 3 mm thick composite built up. Specimens were sectioned into sticks and submitted to a μTBS test or nanoindentation analysis along the interface (24 h or 25,000 thermocycles). The ultramorphology of the polymer–resin interface was also evaluated using CLSM. The results were analyzed using two-way ANOVA and Bonferroni’s post hoc test (α = 0.05). All flavonoids improved short- and long-term μTBS values (p < 0.01), while only some specific such solutions improved the nanomechanical properties (p < 0.05) and preserved the structural morphology of the interface after aging. Pretreatment of acid-etched dentine using specific flavonoid-containing solutions may be a promising approach to improve both the nanomechanical properties and the durability of modern universal adhesive systems. Full article

Bartonella bacilliformis (B. bacilliformis), Bartonella henselae (B. henselae), and Bartonella quintana (B. quintana) are bacteria known to cause verruga peruana or bacillary angiomatosis, vascular endothelial growth factor (VEGF)-dependent cutaneous lesions in humans. Given the bacteria’s association with the dermal niche and clinical suspicion of occult infection by a dermatologist, we determined if patients with melanoma had evidence of Bartonella spp. infection. Within a one-month period, eight patients previously diagnosed with melanoma volunteered to be tested for evidence of Bartonella spp. exposure/infection. Subsequently, confocal immunohistochemistry and PCR for Bartonella spp. were used to study melanoma tissues from two patients. Blood from seven of the eight patients was either seroreactive, PCR positive, or positive by both modalities for Bartonella spp. exposure. Subsequently, Bartonella organisms that co-localized with VEGFC immunoreactivity were visualized using multi-immunostaining confocal microscopy of thick skin sections from two patients. Using a co-culture model, B. henselae was observed to enter melanoma cell cytoplasm and resulted in increased vascular endothelial growth factor C (VEGFC) and interleukin 8 (IL-8) production. Findings from this small number of patients support the need for future investigations to determine the extent to which Bartonella spp. are a component of the melanoma pathobiome. Full article

A double staining protocol for image acquisition using confocal microscopy (CLSM) coupled with image analysis was employed to elucidate the crust and cross-sectional properties of fried dough. Penetrated oil by image analysis (POia), porosity and pore features were quantified from the cross-section micrographs. Crust surface roughness was measured using fractal metrics and fat content was determined by solvent extraction using the American Association of Cereal Chemists method. Crumb porosity ranged between 54.94%–81.84% and reduced (p < 0.05) with bran addition. Crumb pore sizes ranged from 0–475 µm with <1 circularity, indicating elliptical shape. POia values were notably higher (p < 0.05) than PO by Soxhlet extraction (POsox), except for wheat bran (WB) fried dough where the values of POia and POsox were closely ranked. The linear effect of initial moisture content and bran concentration showed a significant impact on the image properties. The mean fractal dimension (FD) decreased as initial moisture increased. The addition of WB caused a significant reduction in the FD of fried dough, while the opposite effect was noted for its oat bran counterpart. Due to non-collinearity of image properties (FD, POia and porosity), data were fitted to cubic polynomial regression with R² values > 0.70. CLSM and image analysis were effective in measuring oil absorption and interpreting crumb properties of fried dough. The protocol used in this study can be applied to other thick deep-fried foods for qualitative observation and quantitative measurement of a specific physical or chemical property. Full article

Oxide layers on titanium foils were produced by galvanostatically controlled plasma electrolytic oxidation in 12.9 M sulfuric acid with small amounts of phosphoric acid added up to a 3% mole fraction. In pure sulfuric acid, the oxide layer is distinctly modified by plasma discharges. As the time of the process increases, rough surfaces with typical circular pores evolve. The predominant crystal phase of the titanium dioxide material is rutile. With the addition of phosphoric acid, discharge effects become less pronounced, and the predominant crystal phase changes to anatase. Furthermore, the oxide layer thickness and mass gain both increase. Already small amounts of phosphoric acid induce these effects. Our findings suggest that anions of phosphoric acid preferentially adsorb to the anodic area and suppress plasma discharges, and conventional anodization is promoted. The process was systematically investigated at different stages, and voltage and oxide formation efficiency were determined. Oxide surfaces and their cross-sections were studied by scanning electron microscopy and energy-dispersive X-ray spectroscopy. The phase composition was determined by X-ray diffraction and confocal Raman microscopy. Full article

Since the resolution of the ANA grade [Amborellales, Nymphaeales, Austrobaileyales] as sister to all other flowering plants, a few comparative studies of root structure have suggested that some of their anatomical traits could be of importance to understanding root evolutionary development and angiosperm phylogeny. However, there is still a paucity of information on root structure and apical meristems (RAMs) in these lineages and especially the sister to all other Austrobaileyales, Austrobaileya scandens. We used microtome sections and bright field, epifluorescence, laser confocal, and scanning electron microscopy to study adventitious root RAMs and tissues of A. scandens. Our results indicate that root structure is relatively simple in A. scandens. The epidermis has a thick cuticle and lacks root hairs. The stele is typically diarch, or some modification thereof, and surrounded by a cortex differentiated into a uniseriate endodermis, a middle region sometimes packed with starch, some oil cells, and colonized by arbuscular mycorrhizal fungi, and a multiseriate exodermis. Secondary growth produced many vessel elements in the secondary xylem and scattered sclerenchymatous fibers in secondary phloem. The absence of distinct patterning within the RAM and between the RAM and derivative differentiating tissues shows that the RAM is open and characterized by common initials. Roots structure and anatomy of A. scandens are thus essentially similar to some previously described in Amborella or Illicium in the ANA grade and many magnoliids, and suggest that the first woody flowering plants likely had an open RAM with common initials. Their functional and evolutionary significance in woody early-diverging and basal lineages of flowering plants and gymnosperms remains unclear, but they are clearly ancestral traits. Full article