Toluidine Blue Solution Research Articles

Sealing ability of various endodontic sealers with or without ethylenediaminetetraacetic acid (EDTA) treatment on bovine root canal.

This study investigated the wettability and consistency of various endodontic sealers, both inorganic and organic, and evaluated their sealing ability of root canals using the single-cone obturation technique, with and without ethylenediaminetetraacetic acid (EDTA) treatment. Bovine root canals were endodontically prepared and filled in preparation for the dye penetration test with toluidine blue solution. All sealers exhibited contact angles similar to or lower than dentin and displayed superior consistency. Among the sealers, organic sealers used without EDTA treatment showed reduced dye penetration compared to inorganic sealers. However, some inorganic and organic sealers showed dye penetration in the sealer and dentin of root canals subjected to EDTA treatment. In conclusion, the single-cone obturation technique, combined with these endodontic sealers, achieved close contact with root canal dentin due to their wettability and consistency. However, the sealing ability of certain sealers was influenced by EDTA treatment.

Community-based early detection of oral precancerous lesion, accessibility and utilization of oral healthcare services among Irular tribes residing at union territory of Pondicherry.

The present study was conducted to estimate the prevalence of oral carcinoma and precancerous lesion, oral healthcare seeking behaviour and identify accessible areas of tobacco cessation centres and oral healthcare facilities among Irular adult tribes residing in Pondicherry. 1. To estimate the prevalence of oral carcinomas and precancerous lesion by using toluidine blue solution. 2. To determine their oral healthcare seeking behaviour in terms of utilizing oral healthcare facilities. 3. To identify accessible tobacco cessation centres and oral healthcare centres for prevention of oral cancer. A cross-sectional study is designed to identify oral carcinomas, oral health-care seeking behaviour among Irular tribes of Pondicherry. 1% toluidine blue solution prepared for detection of oral precancerous lesion. Data was collected using a questionnaire and clinical examination. The collected data was subjected to statistical analysis. Out of 587 study population, oral mucosal lesion among the study population was 46 (7.8%) leukoplakia and 18 (3.1%) candidiasis. Positive health seeking behaviour 24 (41.7%) was observed. The study concluded that low prevalence of oral precancerous lesion and oral healthcare seeking behaviour is poor.

Effect of silver diamine fluoride on the microleakage of flowable resin composite and glass ionomer cement restorations to carious primary dentin: an-in vitro study

BackgroundThis study aimed to evaluate the effect of Silver Diamine Fluoride (SDF) on the microleakage of flowable resin composite (FRC) and resin-modified glass ionomer cement (GIC) restorations bound to carious primary dentin.MethodsForty-four extracted carious primary molars were allocated into four groups as follows (n = 11 teeth/group): Group I, Flowable resin composite (FRCa): SDF38% treatment + FRC, Group II, Flowable resin composite (FRCb): FRC without SDF treatment, Group III, Resin-modified glass ionomer cement (GICa): SDF38% treatment + GIC, Group IV, Resin-modified glass ionomer cement (GICb): GIC without SDF treatment. Specimens were subjected to thermo cycling at 500 cycles between 5 to 55 °C (dwell time of 60 seconds) in baths before being immersed for 24 h in a 1% toluidine blue solution. Microleakage testing was conducted for each specimen in two areas; occlusal and gingival. Specimens were evaluated under stereomicroscope at 4x magnification. Results were analyzed using Kruskal-Wallis test followed by pairwise comparisons utilizing Dunn’s post hoc test at p ≤ 0.05.ResultsInsignificant differences between different groups (p = 0.49) were observed at the gingival walls area readings. The highest value was found in GICb (2.33 ± 0.52), while the lowest value was found in FRCa (1.71 ± 0.76). Insignificant differences between different groups (p = 0.982) were observed at the occlusal walls area readings. The highest value was found in FRCa (1.43 ± 0.98), while the lowest value was found in GICb (1.17 ± 1.33).ConclusionSDF does not adversely affect the microleakage of FRC and GIC restorations bound to carious primary dentin.

Vulvoscopy: anachronism or a time-tested method?

Objective. Due to the growing number of patients with various diseases of the vulva, there is a need to evaluate the available diagnostic tools. Vulvoscopy is one of the widely used methods, the diagnostic sensitivity and specificity of which varies significantly.Purpose of the work. Based on the analysis of publications to determine the diagnostic significance of vulvoscopy.Material and methods. The data of modern research and publications were used in the preparation of the work.Results. The authors of a number of publications warn against considering vulvoscopy as a variant of colposcopy. A high percentage of false positive results when performing tests with acetic acid may be due to a variety of reasons and is not highly specific for papillomavirus infection. The use of the test with toluidine blue is not recommended due to low sensitivity.Conclusions. A positive aspect of vulvoscopy is the ability to examine the vulva area with optical magnification, in a position convenient for examining the patient, which makes it possible to identify areas to be biopsied. The use of additional tests with acetic acid should be limited, and tests with toluidine blue and Lugol solution are not recommended due to low sensitivity.

Physiological and anatomical aspects of the rooting of Brunfelsia pauciflora cuttings

• Cuttings represent a viable alternative for the vegetative propagation of Brunfelsia . • The limited rooting of the cuttings was not associated with an anatomical barrier. • Greater quantities of visibly detected starch in B. uniflora favored its rooting. Brunfelsia is an important genus of the Solanaceae family due to its ornamental aspects and pharmacological properties. Brunfelsia pauciflora stem cuttings represent an alternative to seedling production for its propagation. There is no scientific data currently available concerning B. pauciflora cuttings, although previous studies with Brunfelsia uniflora indicated that it may be difficult to root. Anatomical barriers have been reported as limiting factors for some species that are difficult to root. We investigated the effects of the plant growth regulator indolebutyric acid (IBA) and the influence of the seasons of the year on the rooting of B. pauciflora stem cuttings and compared its rooting to B. uniflora . We also investigated the possible influence of anatomical barriers and starch contents on their rooting. Cuttings of B. pauciflora (7 ± 1 cm long) were made in the autumn of 2019, spring/2019, and summer/2020, and treated with IBA at concentrations of 0, 2000, 4000, and 8000 mg L −1 . For comparison, similar cuttings of B. uniflora were prepared during spring/2019 and treated with IBA concentrations of 0, 4000, and 8000 mg L −1 . The rooting variables of both species were analyzed after 120 days of growth in a greenhouse. For the anatomical analyses, segments of the base of the cuttings were collected on the day of their collection, and again after the 120 day growth period. The samples were embedded in historesin, sectioned, and stained with toluidine blue and Lugol's iodine solution. No combined influence of plant growth regulator concentrations and the season of the year was identified, indicating that those factors act separately on the adventitious rooting of B. pauciflora . The IBA plant growth regulator did not significantly affect any of the variables analyzed. The greatest rooting of B. pauciflora (38.8%), the highest leaf maintenance (82.0%), and the lowest mortality rate (11.2%) were observed in the summer. The species can be classified as difficult-to-root. B. uniflora , on the other hand, evidenced a greater rooting capacity (52.2%) in the spring than B. pauciflora (12.2%). The use of IBA (4000 mg L −1 ) resulted in an increased rooting rate of B. uniflora . Anatomically, it was possible to identify the origin of the root primordia as the vascular cambium region, with root formation being defined as direct. A ring of perivascular fibers, often indicated in the literature as an anatomical barrier, was not observed as such in B. pauciflora , as rooting not only overcame the fibers, but also the callus mass. The two species of Brunfelsia differed in terms of their starch contents, which was greater in B. uniflora and appeared to favor its rooting.

An Experimental Study of Photoactivated Disinfection in the Treatment of Acute Pseudomembranous Stomatitis.

To investigate photoactivated disinfection (PAD) to treat acute pseudomembranous stomatitis, an animal model was established. Six-week-old male ICR mice were inoculated with Candida albicans under immunosuppression then divided into three groups (15 mice per group). Pseudomembranous area was measured, then mice had 1 mg mL-1 toluidine blue solution spread on the area, left for 1 min (PAD-1 group) or 2 min (PAD-2 group), then irradiated with a 750 mW LED red light for 1 min, a control group received no treatment. Fungal load was measured immediately; after 48 h of observation pseudomembranous area and fungal load were measured. The mice were sacrificed and histopathological examination was performed. Before treatment, pseudomembranous area scores were similar (3 to 4 points) in all groups; 48 h after treatment, the treatment groups' scores were lower (1 to 2 points) than the control group (3 to 4 points, P < 0.05). Immediately after treatment and 48 h later, the fungal loads of the treatment groups were lower than the control group (both P < 0.05). Histopathology of the treatment groups improved more than controls. The treatment groups' results were similar. Therefore, this method of PAD, with short treatment time, reduced the fungal load and pseudomembranous area scores in a mouse model of acute pseudomembranous stomatitis and may have clinical potential.

IMPROVEMENT OF AUTOFLUORESCENCE IMAGING METHOD IN DETECTION OF CANCEROUS LESIONS OF THE ORAL MUCOSA

Purpose: to optimize the diagnostic method for detecting neoplasms of the oral mucosa. Materials and methods: The study group included 32 people with lesions of the oral mucosa: lichen planus — 17; chronic mechanical injury (decubital ulcer) — 15. The authors have proposed a method for diagnosing erosive and ulcerative elements of the oral mucosa, which consists in staining pathological foci with a 1% solution of tuloidine blue followed by autofluorescence imaging. The effectiveness of autofluorescence imaging by «AFS» stomatoscope, staining and a combination of two methods in visualizing the boundaries of precancerous lesions of the oral mucosa was determined. Results and discussion: The technique of autofluorescence imaging by «AFS» stomatoscope, in 63.7% incorrectly diagnosed patients with cancer of the oral mucosa, in addition, 27.3% did not reveal malignant neoplasms. Marking the lesion elements with a 1% toluidine blue solution and their illumination with autofluorescence imaging, made it possible to establish a dark glow zone in those cases (27.3%) when imaging did not identify the pathological focus as an area without fluorescence. In all patients with malignant neoplasms of the oral mucosa diagnosed by autofluorescence imaging with staining, a truly positive result was confirmed by morphometric data. Conclusion: Autofluorescence imaging with staining has a high sensitivity and specificity of 100%, and is 2 times more effective than autofluorescence imaging in the diagnosis of dysplasia of erosive and ulcerative elements of the oral mucosa.

Toluidine blue for the detection of sentinel lymph nodes in patients with thyroid cancer

Thyroid cancer (TC) demonstrates steady growth in incidence worldwide and remains an urgent problem in oncology. The detection of sentinel lymph nodes (SLN) with a selective dye and further histological examination in selecting the proper (personalized) surgical strategy and the volume of surgical intervention for clinically undetermined regional lymph nodes. The purpose of the study is to evaluate the effectiveness and safety of intraoperative detection of SLN with a 1% toluidine blue aqueous solution. The significant tasks are to identify the pattern of TC metastases to cervical lymph nodes, to establish the prevalence of "skip" metastases, to compare the frequency of complications after total thyroidectomy and central neck dissection and lateral neck dissection with total thyroidectomy and central neck dissection, and to determine the feasibility of the application of lateral neck dissections in patients with papillary and follicular TC without metastases to regional lymph nodes (according to physical examination and ultrasound). According to our data the SLN identification rate was 97.6%. Sensitivity, specificity, positive predictive value, negative predictive value, and frequency of false negative and false positive results was 89.2, 94.6, 88.03, 95.16, 10.8, and 5.4%, respectively. The most common metastasis was in the central neck compartment (83.7%). Skip metastases were determined in 4.9% of patients. The low prevalence of "skip" metastases and a significant risk of postoperative complications (wound exudation, lymphorrhagia, inflammation, hypoparathyroidism, paresis of the vocal cords) support the idea that lateral neck dissection is appropriate only in cases of confirmed metastases by physical examination and/or ultrasound at the preoperative stage.

Fine-Tuning of Molecular Structures to Generate Carbohydrate Based Super Gelators and Their Applications for Drug Delivery and Dye Absorption.

Carbohydrate-based low molecular weight gelators (LMWGs) exhibit many desirable properties making them useful in various fields including applications as drug delivery carriers. In order to further understand the structural connection to gelation properties, especially the influence of halide substitutions, we have designed and synthesized a series of para-chlorobenzylidene acetal protected D-glucosamine amide derivatives. Fifteen different amides were synthesized, and their self-assembling properties were assessed in multiple organic solvents, as well as mixtures of organic solvents with water. All derivatives were found to be gelators for at least one solvent and majority formed gels in multiple solvents at concentrations lower than 2 wt%. A few derivatives rendered remarkably stable gels in aqueous solutions at concentrations below 0.1 wt%. The benzamide 13 formed gels in water and in EtOH/H2O (v/v 1:2) at 0.36 mg/mL. The gels were characterized using optical microscopy and atomic force microscopy, and the self-assembly mechanism was probed using variable temperature 1H-NMR spectroscopy. Gel extrusion studies using H2O/DMSO gels successfully printed lines of gels on glass slides, which retained viscoelasticity based on rheology. Gels formed by the benzamide 13 were used for encapsulation and the controlled release of chloramphenicol and naproxen, as well as for dye removal for toluidine blue aqueous solutions.

ELECTRONIC-MICROSCOPIC CHANGES OF THE INTRAMURAL NERVOUS PLEXUS OF THE DUODENUM IN CONDITIONS OF CHRONIC STRESS

The article presents a study of ultrastructural changes of neurons of the intramural nervous system (muscular- intestinal plexus) of the duodenum under chronic stress. The experiment was performed on 60 white adult male rats. The material was taken from the proximal and distal parts of the duodenum after 14, 28, 56 days of the experiment. For electron microscopic examination, pieces of the duodenum were fi xed in 2.5% glutaraldehyde solution for 4 hours, and then fi xed in 2% osmium tetroxide solution. First, semi-thin sections were made to detect the localization of nerve cells of the musculoskeletal plexus of the duodenum, staining them in 1% methylene blue solution or 0.1% toluidine blue solution. Ultrathin sections were made on an ultramicrotome Tesla BS-490A, the material was studied on an electron microscope TEM – 125 K («Selmi», Ukraine). After 14 days of chronic stress, there is signifi cant damage to the neurons of the musculoskeletal plexus of the duodenum due to reactive and edematous processes, accompanied by the development of ultrastructural changes and the death of some neurons. After 28-56 days of the experiment, edematous, dystrophic- destructive processes in nerve cells are further observed, but in some of them there are a number of compensatory- adaptive and reparative processes aimed at restoring impaired functions.

Optimization of Toluidine Blue Biosorption in Aqueous Solutions Using Polyporus Squamosus Fungi as Absorbent by Response Surface Methodology

Textile wastewater including a large number of dyes and heavy metals can have adverse impacts on human health and surface water. In this work, biosorption Toluidine Blue from aqueous media onto natural Polypourus squamosus fungi as a low-cost biosorbent was investigated. Central Composite Design (CCD) in Response Surface Methodology (RSM) was successfully applied to optimize the biosorption condition. Medium parameters affected the biosorption of Toluidine Blue were determined to be initial pH, initial Toluidine Blue (Tb) concentration, temperature, and absorbent dosage. All experiments were carried out in a batch system using 250 mL flasks containing 100 mL of Toluidine Blue solution with a temperature-controlled magnetic stirrer. The Tb concentrations remaining in filtration solutions after biosorption were analyzed using UV-Spectro. With the obtained quadratic model, the optimal conditions for maximum biosorbed Toluidine blue were calculated to be 7, 27.5 mg/L, 35°C and 0.05 g for pH, C°, T (°C) and adsorbent dosage, respectively. Furthermore, most known isotherm models such as Langmuir and Freundlich were computed to find the best-fitted model.

DETECTION OF SENTINEL LYMPH NODES IN PATIENTS WITH THYROID CANCER WITH THE USE OF TOLUIDINE BLUE

Introduction. Treatment of patients with thyroid cancer (TС) with clinically undetected metastases to regional lymph nodes remains an unsolved problem of modern oncology. The search for reliable diagnostic methods, which underlie the decision to perform an adequate lymphadenectomy for TC, is one of the priorities of oncosurgery. The aim of the research: to find an effective method of intraoperative contrasting of sentinel lymph nodes (SLN) in patients with TC using toluidine blue and to evaluate its effectiveness. Methods. A study of 123 patients with TC who underwent surgical treatment using the contrast-visual method in the amount of thyroidectomy with central and bilateral selective cervical lymph node dissection (levels VI; IIA; III; IV). During surgery, a 1% solution of toluidine blue was injected into the tumor using a syringe. Surgical treatment was performed in two stages. At the first stage – thyroidectomy was performed in a block with a central lymphatic collector of the neck (level IV), which included removal of paratracheal, prelaryngeal, parathyroid tissue of the neck. Urgent intraoperative histological examination of thyroid tumor and distant SLN was performed. In the second stage of the operation performed cervical lymph node dissection with removal of fiber IIA; III; IV levels of the neck, with mandatory revision of the level of VB. After the operation, the final histological examination of the removed specimen with the tumor and all lymph nodes was performed. Results. After the application contrasting technique, SLN were found in 120 (97.6%) patients. Their number varied from 2 to 12. These lymph nodes were mainly localized in the central collector of the neck on the side of the primary tumor (central, ipsilateral SLN) in 91 (75.8%) patients. Based on the results of an intraoperative study of SLN, metastases were found in 33 (27.5%) of 120 patients. In most cases (83.3%) SLNs were detected in the central lymphatic collector of the neck (level VI). In 76 (63.3%) patients, staining of the lymph nodes of the III level of the neck (middle ipsilateral jugular lymphatic collector) occurred, in 11 (9.2%) SLNs were found only at levels III, IV and VB of the neck. Very rarely (5.8%), the lymph nodes of the anterior-superior mediastinum were contrasted. Conclusions. The method of contrast visualization of the regional collector in patients with TC using a 1% solution of toluidine blue is a highly informative study that allows to determine the regional lymph nodes affected by metastases and to differentiate the choice of the volume of lymph node dissection in the early stages of tumors. The method is simple to implement, does not involve considerable costs and can be recommended for use in oncology clinics.

Effect of sealing gel on the microleakage resistance and mechanical behavior during dynamic loading of 3 implant systems

Statement of problemSealing products have been produced to reduce microleakage at the implant abutment interface. However, little is known about their effectiveness and any alterations in mechanical behavior of implant systems with their application. PurposeThe purpose of this in vitro study was to evaluate the effect of a silicone sealing gel on implant abutment interface microleakage, abutment screw torque loss, and thread wear of implant systems in a simulated oral environment. Material and methodsFive specimens each of 3 implants systems (Nobel, Straumann, and WEGO) that included sealed and unsealed groups were analyzed (N=30). Before assembling the components, toluidine blue solution was injected to the implant intaglio cavity to evaluate implant abutment interface microleakage. After tightening to the recommended torque, 20 to 200 N of 30-degree off-axis dynamic force was applied at 2 Hz for 48 hours. The toluidine blue solution was extracted to test optical density values at 1, 3, 9, 24, 33, and 48 hours. Detorque values were measured before and after cycling loading, and torque loss rates were calculated. The abutment screw morphologies were observed by using scanning electron microscopy. The coefficient of friction tendency of applying sealing gel was explored with a ball-on-flat configuration. One-way ANOVA and Student t test were used for statistical analysis (α=.05). ResultsThe optical density value increased with the loading time, especially for Straumann group. The sealing gel decreased the implant abutment interface microleakage of Straumann assemblies after cyclic loading of 9 hours (P=.044), whereas no statistical difference was found for Nobel (P=.140) or WEGO groups (P=.402) at 6 time points. Torque loss occurred during tightening and further increased after dynamic cyclic load in each group. Among the 3 implant systems, Straumann implants reported the best antiloosening property (P<.001). The application of sealing gel reduced the initial (P=.048) and final (P=.032) torque loss rate in all the 3 systems. Scanning electron microscopy observations revealed the bottom thread tended to have more abrasion than the first thread. After applying sealing gel, less thread abrasion was found in Nobel and WEGO assemblies, whereas the protective effect was not evident for the Straumann group. The coefficient of friction of sealed group (0.17 ±0.026) was significantly (P=.012) lower than that of unsealed group (0.24 ±0.044). ConclusionsThe silicone sealing gel improved the immediate fastening and long-term antiloosening performances of 3 implant systems, decreased the implant abutment interface microleakage of Straumann system, and reduced abutment screw thread abrasion of the Nobel and WEGO systems.

Role of mast cells in aetiopathogenesis of radicular cyst

Role of mast cells in aetiopathogenesis of radicular cyst - JOOO- Print ISSN No: - 2395-6186 Online ISSN No:- 2395-6194 Article DOI No:- 10.18231/j.jooo.2020.019, Journal of Oral Medicine, Oral Surgery, Oral Pathology and Oral Radiology-J Oral Med Oral Surg Oral Pathol Oral Radiol

A novel nucleic acid based bio-assay toward recognition of Haemophilus influenza using bioconjugation and DNA hybridization method

Haemophilus influenza (H. influenza) is a gram negative coccobacillus pathogenic microorganism. H. influenza produces beta-lactamases, and it is also able to modify its penicillin-binding proteins, so it has gained resistance to the penicillin family of antibiotics. In this work, a novel sensitive approach was established for the monitoring of H. influenza using DNA based bio-assay. For the first time, specific sequence of thiolated probe of Haemophilus influenza (SH-5′-AAT TTT CCA ACT TTT TCA CCT GCA T-3′) was immobilized on the surface of gold (Au) electrode. Square wave voltammetry (SWV) was carried out in toluidine blue (TB) solution for DNA hybridization and targeting of cDNA sequence of Haemophilus influenza. Field scanning electron microscope (FE-SEM) was applied to investigation of the electrode morphology and estimate of particle size. In the optimal conditions, the planned strategy could detect target DNA (5′-ATG CAG GTG AAA AAG TTG GAA AAT T-3′) down to 1 ZM with a linear range from 1 μM to 1 ZM. Moreover, engineered geno-assay selectively differentiates the complementary sequence from target sequences with one, double and three base mismatch sequences.

Possible adaptation of autologous osteochondral regenerative medicine using umbilical cord-derived mesenchymal stromal cells of cleft palate patient

Background & Aim Cleft palate is a major obstacle to constraining ADL for the children. In this study, to regenerate and compensate the defected palate by autologous umbilical cord-derived mesenchymal stromal cells (UC-MSC) of cleft palate patient, we studied the osteochondrogenic differentiation potency of UC-MSC. Methods, Results & Conclusion Materials/Methods UC-MSC derived from a patient who had cleft palate (CLP-UC-MSC) at the time of birth was obtained from the umbilical cord blood and cord bank of the Institute of Medical Science, The University of Tokyo (IMSUT CORD). We also obtained MSC derived from two healthy children (H-UC-MSC) as control and compared them. Banking and the experiments of this study were approved by IRB of IMSUT. Result The proliferation and surface markers of CLP-UC-MSC (CD105 + D73 + CD90 + HLA class I + CD45-HLA-DR-) were equivalent to H-UC-MSC. Expanded CLP-UC-MSC at passage 4 (P4) showed normal karyotype. Osteogenic differentiation using P4 cells was performed by seeding 12-well plates in 1 ml of the osteogenic differentiation medium, replacing the medium every 5 days, and culturing for 30 days. In Alizarin Red S staining, deposited Ca was confirmed in differentiated CLP-UC-MSC as well as those in the H-UC-MSC. Chondrogenic differentiation potency using P4 cells was evaluated by a pellet method in 15ml centrifuge tube made of PP, adding chondrogenic differentiation medium. After 24 days of culture, intracellular mucoitinsulfuric acid and chondroitin sulfate stained by Toluidine Blue Solution was observed. Discussion In this study, we demonstrated that CLP-UC-MSC showed the equivalent osteochondrogenic differentiation potential to H-UC-MSC, although only one case of cleft palate was examined. It was reported previously that UC-MSC showed less osteogenic differentiation ability compared to bone marrow-derived MSC, but we succeeded to induce the osteogenic differentiation potency, suggesting that it could be applied to osteochondral regeneration using autologous CLP-UC-MSC and H-UC-MSC.

Applicable possibility of autologous osteochondral regenerative medicine using umbilical cord-derived mesenchymal stromal cells of cleft palate patient

Background & Aim Cleft palate is a major obstacle to constraining ADL for the children. In this study, to regenerate and compensate the defected palate by autologous umbilical cord-derived mesenchymal stromal cells (UC-MSC) of cleft palate patient, we studied the osteochondrogenic differentiation potency of UC-MSC. Methods, Results & Conclusion Materials/Methods UC-MSC derived from a patient who had cleft palate (CLP-UC-MSC) at the time of birth was obtained from the umbilical cord blood and cord bank of the Institute of Medical Science, The University of Tokyo (IMSUT CORD). We also obtained MSC derived from two healthy children (H-UC-MSC) as control and compared them. Banking and the experiments of this study were approved by IRB of IMSUT. Result The proliferation and surface markers of CLP-UC-MSC (CD105 + D73 + CD90 + HLA class I + CD45-HLA-DR-) were equivalent to H-UC-MSC. Expanded CLP-UC-MSC at passage 4 (P4) showed normal karyotype. Osteogenic differentiation using P4 cells was performed by seeding 12-well plates in 1 ml of the osteogenic differentiation medium, replacing the medium every 5 days, and culturing for 30 days. In Alizarin Red S staining, deposited Ca was confirmed in differentiated CLP-UC-MSC as well as those in the H-UC-MSC. Chondrogenic differentiation potency using P4 cells was evaluated by a pellet method in 15ml centrifuge tube made of PP, adding chondrogenic differentiation medium. After 24 days of culture, intracellular mucoitinsulfuric acid and chondroitin sulfate stained by Toluidine Blue Solution was observed. Discussion In this study, we demonstrated that CLP-UC-MSC showed the equivalent osteochondrogenic differentiation potential to H-UC-MSC, although only one case of cleft palate was examined. It was reported previously that UC-MSC showed less osteogenic differentiation ability compared to bone marrow-derived MSC, but we succeeded to induce the osteogenic differentiation potency, suggesting that it could be applied to osteochondral regeneration using autologous CLP-UC-MSC and H-UC-MSC.

Microleakage at Different Implant-Abutment Connections Under Unloaded and Loaded Conditions.

This study evaluated the microleakage at different implant-abutment (I-A) connections under unloaded and loaded conditions. Forty implants, specially designed with an opening at the apex, were grouped according to the I-A and screw device: external hexagon implants with titanium (EH) or EH diamond-like carbon screws fixing the abutment; internal hexagon implants with titanium screws (IH); and Morse taper implants with solid (MT) or MT passing screws (MTps) abutments. The implants were fixed in a 2-compartment device, and toluidine blue solution (1.0 mg/mL) was placed at the I-A interface (upper compartment). The lower compartment was filled with purified water. Four implants of each group were loaded (50 N, 1.2 Hz) and after 50,000, 100,000, 200,000, and 300,000 cycles, aliquots were collected from the lower compartment for absorbance reading. Data were analyzed using analysis of variance for repeated measurements and Tukey (α = 0.05). Although microleakage increased over time in most of the groups, MTps group showed lower values when loaded (P < 0.05). The DLC on the EH screw did not prevent microleakage (P < 0.05). It can be concluded that MT connection is more effective to prevent microleakage.

Determination of carrageenan by means of photometric titration with Methylene Blue and Toluidine Blue dyes

A new approach to carrageenan quantitation was described. The method consists in titration of carrageenan solution by Methylene Blue dye or Toluidine Blue dye solution until a certain absorbance value. The optimal wavelengths are 660nm and 640nm when titrating with Methylene Blue dye and Toluidine Blue dye, respectively. Rectilinear calibration plots (R2>0.996) provide carrageenan determination in the concentration range from 2 to 60mg/L with relative standard deviation from 1 to 5%. The proposed method is simple and feasible in use due to optical dip probe providing in situ absorbance measurements. The proposed way of end-point recognition as pre-set voltage is applicable with any automatic titrator. The method was tested on model jelly dessert sachet. No interference was registered from typical ingredients of jellies such as sucrose, citric acid, sodium citrate, malic acid, potassium sorbate as well as blue colorant. The neutral polysaccharides such as guar gum and locust bean gum have insignificant interference when their content is fivefold as compared to that of carrageenan.

Histochemical observation of bony reversal lines in bisphosphonate-related osteonecrosis of the jaw

To contrast the pattern of bony reversal linesin bisphosphonate osteonecrosis of the jaw with infected osteomyelitis derived acute osteonecrosis of the jaw. This study investigated the histochemical characteristics of reversal lines in 50 cases of BP-related osteonecrosis of the jaw (BRONJ) compared with non-BP-involved bones in 20 cases of chronic osteomyelitis of the jaws. Necrotic bones were stained by using the toluidine blue, Safranin O, Giemsa, van Gieson, and Masson's trichrome staining methods. All BP-involved bones in BRONJ were distinguishable from non-BP-involved bones in chronic osteomyelitis of the jaws by multiple thick, irregular, reversal lines, which were strongly stained with toluidine blue, Safranin O, and Giemsa solution. The reversal lines of BP-involved bones (average 31.2±10.85μm) were thicker than those of osteomyelitic bones (average 11.1±3.76μm), and they were closely associated with immature bony matrices containing collagenous materials positive for van Gieson and Masson's trichrome staining with statistical significance (P=.0212 in t test statistics). The immature reversal lines of BP-involved bones continuously appeared as thick non-birefringence lines between lamellate structures as observed under a polarizing microscope, whereas the reversal lines of non-BP-involved bones were gradually thinned as their mineralization advanced. BP-involved bones had immature bony matrices outlined by thick reversal lines, which might be crucial to rapid osteonecrosis of BRONJ and also could be hallmarks for the differential diagnosis of BRONJ from chronic osteomyelitis of the jaws.