Search

Aim: to examine the implantation of chitosan channels stuffed with mesenchyme-originated stem/progenitor cells (MSPCs) derived from adult rats in a spinal cord transection model. The level of axonal regeneration, the effect of chitosan channels on the survival of MSPCs, and the functional recovery results were also evaluated. Material and methods: chitosan channels stuffed with MSPCs were implanted at the level of T8 in a transected rat spinal cord. MSPCs were harvested from the pelvic bone marrow of adult rats, and the MSPC-chitosan channel group was compared with three control groups. The axonal regeneration capacity, the effect of chitosan channels on the survival of MSPCs, and the functional recovery results were compared among four groups. The survival of MSPCs was evaluated using histopathological techniques and electron microscopy, axonal regeneration/germination was evaluated by confocal microscopy, and locomotor function was assessed for 4 weeks using the Basso, Beattie, and Bresnahan locomotor score. Results: the MSPC-chitosan channel group exhibited enhanced survival of transplanted MSPCs compared with MSPCs transplanted directly into the lesion cavity, although no significant difference was detected in locomotor function between the treatment and control groups. The MSPC-chitosan channel group demonstrated thicker myelination of axons than the other groups. Conclusion: chitosan channels promoted the survival of transplanted MSPCs and created a tissue bridge after complete spinal cord transection. They also induced axonal regeneration and germination. No significant improvement in functional recovery was found between the groups., Hacettepe University

This study investigated the dentinal tubule penetration of mineral trioxide aggregate (MTA), NeoMTA Plus and Biodentine placed by either manual condensation or ultrasonic activation in simulated open apex model. Standardized divergent open apex models were created using palatal roots of 60 human maxillary molars and divided into six groups according to the used cements and activation methods (n = 10): MTA-manual condensation, MTA-ultrasonic activation, NeoMTA Plus-manual condensation, NeoMTA Plus-ultrasonic activation, Biodentine-manual condensation, Biodentine-ultrasonic activation. For the measurement of penetration, the cements were mixed with 0.1% Rhodamin B and 6-mm apical portions of each root canal were obturated in an orthograde direction. The roots were embedded into acrylic blocks, and 1-mm-thick sections were obtained at 3 mm from the apex. Specimens were mounted onto glass slides and scanned under a confocal laser scanning microscope (CLSM) and stereomicroscope. Dentinal tubule penetration areas, depth and percentage were measured using LSM and ImageJ software. The data were analyzed using two-way analysis of variance (anova) with Bonferroni correction (α = 0.05). No correlation was found between stereomicroscope and CLSM analyses (p > .05). CLSM analysis showed no significant differences between MTA, NeoMTA Plus, and Biodentine groups when manual condensation was used (p > .05). Ultrasonic activation did not increase the tubular penetration of MTA, NeoMTA Plus or Biodentine as compared to manual condensation of each material (p > .05). MTA, NeoMTA Plus and Biodentine showed similar tubular penetration when manual condensation was used. Ultrasonic activation of these cements had no effect on tubular penetration of each material as compared to the manual condensation counterparts.

Background.This study compared the effect of smear layer on the penetration depth and push-out bond strength of various root canal sealers. Methods.A total of 90 extracted human mandibular premolars were assigned into 2 groups: smear layer preserved and smear layer removed. Then the roots were further divided into 3 subgroups according to the sealer tested: AH 26, BioRoot RCS and MTA Plus. Obturation was performed with gutta-percha and the relevant sealer was mixed with 0.1% rhodamine B. Three 1-mm-thick slices were obtained from the mid-third area of each root. Two slices were selected for the push-out test and the remaining slice was used to calculate the dentinal tubule penetration depth and percentage. Results.The retention of MTA Plus and BioRoot RCS was higher than that of AH 26 when the smear layer was preserved (P

WOS: 000431796100001, PubMed: 29748557, Amelogenin (AMG) is a cell adhesion molecule that has an important role in the mineralization of enamel and regulates events during dental development and root formation. The purpose of the present study was to investigate the effects of recombinant human AMG (rhAMG) on mineralized tissue-associated genes in cementoblasts. Immortalized mouse cementoblasts (OCCM-30) were treated with different concentrations (0.1, 1, 10, 100, 1000, 10,000, 100,000 ng . mL(-1)) of recombinant human AMG (rhAMG) and analyzed for proliferation, mineralization and mRNA expression of bone sialoprotein (BSP), osteocalcin (OCN), collagen type I (COL I), osteopontin (OPN), runt-related transcription factor 2 (Runx2), cementum attachment protein (CAP), and alkaline phosphatase (ALP) genes using quantitative RT-PCR. The dose response of rhAMG was evaluated using a real-time cell analyzer. Total RNA was isolated on day 3, and cell mineralization was assessed using von Kossa staining on day 8. COL I, OPN and lysosomal-associated membrane protein-1 (LAMP-1), which is a cell surface binding site for amelogenin, were evaluated using immunocytochemistry. F-actin bundles were imaged using confocal microscopy. rhAMG at a concentration of 100,000 ng . mL(-1) increased cell proliferation after 72 h compared to the other concentrations and the untreated control group. rhAMG (100,000 ng . mL(-1)) upregulated BSP and OCN mRNA expression levels eightfold and fivefold, respectively. rhAMG at a concentration of 100,000 ng . mL(-1) remarkably enhanced LAMP-1 staining in cementoblasts. Increased numbers of mineralized nodules were observed at concentrations of 10,000 and 100,000 ng . mL(-1) rhAMG. The present data suggest that rhAMG is a potent regulator of gene expression in cementoblasts and support the potential application of rhAMG in therapies aimed at fast regeneration of damaged periodontal tissue., TUBITAK (Turkey)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [SBAG108S265]; BMBF (Germany)Federal Ministry of Education & Research (BMBF) [TUR08/09], This study was supported by TUBITAK SBAG108S265 (Turkey) and BMBF TUR08/09 (Germany). We thank Johan Svensson and Leif Bulow for rhAMG production and characterization studies from Department of Pure and Applied Biochemistry, Center for Chemistry and Chemical Engineering Lund University, Lund, Sweden for rhAMG synthesis.

Introduction The purpose of this study was to evaluate dentinal tubule penetration (DTP) of calcium hydroxide (CH) and triple antibiotic paste (TAP) when performed with distilled water (DW) or a low surface tension liquid (ie, propylene glycol [PG]). Methods Root apices of 40 single-rooted premolars were removed to obtain 14-mm roots in length. Root canals were enlarged to simulate immature teeth. After smear layer removal, the roots were randomly divided into 4 groups ( n = 10) according to the root canal medicaments and the vehicles used: group 1:TAP + DW, group 2: TAP + PG, group 3: CH + DW, and group 4:CH + PG. Root canal medicaments were labeled with 0.1% rhodamine and applied into the canals using a Lentulo spiral. Specimens were molded into acrylic blocks, and 1-mm-thick sections were obtained from the middle third of each root. Specimens were mounted onto glass slides and scanned under a confocal laser scanning microscope. DTP depth, percentage, and area were measured using imaging software. Kruskal-Wallis and Mann-Whitney U tests were used for statistical analysis. The level of significance was set at P Results No significant difference was found among the experimental groups in terms of both percentage and depth of DTP ( P > .05). CH had a lower penetration area compared with TAP regardless of the vehicle used ( P Conclusions A low surface tension vehicle did not alter the penetration of CH and TAP.

In order to improve the immunogenicity of the highly purified vaccine antigens, addition of an adjuvant to formulation, without affecting the safety of the vaccine, has been the key aim of the vaccine formulators. In recent years, adjuvants which are composed of a delivery system and immunopotentiators have been preferred to induce potent immune responses. In this study, we have combined Salmonella Typhi porins and chitosan to develop a new adjuvant system to enhance the immunogenicity of the highly purified antigens. Cationic gels, microparticle (1.69 +/- 0.01 mu m) and nanoparticles (337.7 +/- 1.7 mu m) based on chitosan were prepared with high loading efficiency of porins. Cellular uptake was examined by confocal laser scanning microscopy, and the macrophage activation was investigated by measuring the surface marker as well as the cytokine release in vitro in J774A.1 macrophage murine cells. Porins alone were not taken up by the macrophage cells whereas in combination with chitosan a significant uptake was obtained. Porins-chitosan combination systems were found to induce CD80, CD86 and MHC-II expressions at different levels by different formulations depending on the particle size. Similarly, TNF-alpha and IL-6 levels were found to increase with porins-chitosan combination. Our results demonstrated that combination of porins with chitosan as a particulate system exerts enhanced adjuvant effect, suggesting a promising adjuvant system for subunit vaccines with combined immunostimulating activity.

The aim of this study was 2-fold: to evaluate the penetration of a tricalcium silicate-based endodontic sealer (EndoSequence BC Sealer; Brasseler USA, Savannah, GA) into dentinal tubules without a core material (sealer) or with .02 or .04 tapered bioceramic gutta-percha points and to compare the time required to remove the root fillings .Roots of extracted human mandibular incisors (N = 60) were prepared with 0.04 taper nickel-titanium rotary files to #35 and randomly assigned into 3 groups (n = 10/group) according to the obturation method used: 1. obturating with sealer only, 2. sealer + .02 point, and 3. sealer + .04 point. The sealer was labeled with rhodamine B for analyzing dentinal tubule penetration under a confocal laser scanning microscope. The remaining specimens (n = 30) were used to measure the time for removal of the root canal fillings with retreatment files. The data were analyzed using 1-way analysis of variance and post hoc Games-Howell tests for dentinal tubule penetration and the Kruskal-Wallis test for retreatment time.Significantly greater sealer penetration and sealer-penetrated area was achieved when the sealer was used with a .04 gutta point (P.05), whereas there was no difference between the sealer and .02 gutta point groups (P.05). All test groups showed a similar depth of sealer penetration (P.05). Groups with the gutta-percha points required a similar time to remove root filling (P.05), whereas the working length could not be achieved in the sealer group.The use of a matched-taper bioceramic gutta-percha point enhanced the dentinal tubule penetration of the tested tricalcium silicate-based sealer. The use of a core material in conjunction with sealer facilitates removal of the root filling to the working length.

Crayfish is a common model animal for different experimental purposes. However, the lack of information about the genetic properties of the animal limits its use in comparison to other model animals. In the present study, a putative crayfish sodium/calcium exchanger gene has firstly been cloned in ganglia cDNA samples by conducting a series of PCR experiments, where a set of degenerate and specific primers and RACE method were used. The complete sequence is 2955 bp, and the ORF is 2718 bp in length. Molecular properties of the calculated peptide were similar to the sodium/calcium exchangers reported in the other species. Analysis of the qPCR data indicated that the putative gene has the highest expression level in the ganglia. However, an apparently elevated level of expression is observed in highly active tissues like heart, muscle and intestine, while the least expression level was observed in the stomach samples. It was proposed that the cloned gene may code the sodium/calcium exchanger protein in the crayfish.

In the present study, cytosolic calcium concentration changes were recorded in response to various forms of excitations, using the fluorescent calcium indicator dye OG-BAPTA1 together with the current or voltage clamp methods in stretch receptor neurons of crayfish. A single action potential evoked a rise in the resting calcium level in the axon and axonal hillock, whereas an impulse train or a large saturating current injection would be required to evoke an equivalent response in the dendrite region. Under voltage clamp conditions, amplitude differences between axon and dendrite responses vanished completely. The fast activation time and the modulation of the response by extracellular calcium concentration changes indicated that the evoked calcium transients might be mediated by calcium entry into the cytosol through a voltage-gated calcium channel. The decay of the responses was slow and sensitive to extracellular sodium and calcium concentrations as well as exposure to 1–10 mM NiCl2 and 10–500 µM lanthanum. Thus, a sodium calcium exchanger and a calcium ATPase might be responsible for calcium extrusion from the cytosol. Present results indicate that the calcium indicator OG-BAPTA1 might be an efficient but indirect way of monitoring regional membrane potential differences in a single neuron.

This study compared several irrigation protocols and application systems for sealer penetration into dentinal tubules. Single-rooted-human teeth were divided into 5 experimental groups (n = 15) and a control group (n = 5), according to final irrigation protocols: standard needle irrigation (SNI); Vibringe; Vibringe + NaviTip FX (Vibringe NFX); Endo Spray (ES); and passive-ultrasonic-irrigation (PUI). Following obturation of the root canals, the percentage of the sealer penetration was measured at different depths using stepwise CLSM analysis. The sealer penetration in the experimental groups was significantly higher than the control group at all levels (p .05). No significant differences were observed between Vibringe and SNI or Vibringe NFX, ES, and PUI at all depths (p .05). The Vibringe NFX, ES, and PUI groups allowed deeper sealer penetration than SNI at 100, 250, and 500 μm levels (p .05). The irrigant activation, the needle design, and the application form (syringe or spray) may impact the quality of the seal that is achieved with root canal filling.

Objective: Relevancy of the reference genes have not been investigated in crayfish tissue samples. Various genes have been proposed for establishing a reference level to analyze qPCR results obtained in different tissue samples. However, majority of those genes have been investigated solely in mammalian species. There is no report related to the relevancy and the efficiency of those common reference genes in crayfish tissue samples yet. The primary aim of the present work is to compare the expression level of a set of reference genes in various crayfish tissue samples to establish a reliable reference gene for quantitative expression level comparisons.Methods: cDNA copies were synthesized by RT-PCR from equalised total RNA samples of nine different tissue samples. qPCR were performed on cDNA samples by using four different primer pairs specific for β-actin, Peptidylprolyl Isomerase B, Sarcoplasmic Calcium Binding Protein and 18S ribosomal RNA genes.Results: qActin2 amplified gene fragments for β-actin in all samples. Cycle time of the amplification reaction differed substantially between the investigated tissue types. A similar difference was observed between the melting curves of the products. The primer pair PPIB amplified gene fragments for Peptidylprolyl Isomerase B only in heart, muscle and intestine samples but not in the rest of the tissue samples. Further, both amplification and melting curves for heart, muscle and intestine were different. qSCP amplified some gene fragments for Sarcoplasmic Reticulum Binding Protein only in muscle, heart and ganglion samples. The 18S_1 amplified fragments for 18S. Amplification curves the samples superimposed on each other. Melting curves of the products were very similar. Variance of the obtained data was the lowest for 18S ribosomal RNA.Conclusion: In refer to present results; it is conceivable to propose that among all the genes investigated in the present study, expression level of ribosomal 18S RNA is the most homogeneously distributed one within the body parts of the crayfish. Further, it is minimally affected by extreme biological changes such as molting. Thus, 18S Ribosomal RNA would be suggested as a reference gene to be used when investigating tissue specific expression level of a gene in the crayfish.

Objectives The aim of this study was to compare the push-out bond strength and dentinal tubule penetration of root canal sealers used with coated core materials and conventional gutta-percha. Materials and Methods A total of 72 single-rooted human mandibular incisors were instrumented with NiTi rotary files with irrigation of 2.5% NaOCl. The smear layer was removed with 17% ethylenediaminetetraacetic acid (EDTA). Specimens were assigned into four groups according to the obturation system: Group 1, EndoRez (Ultradent Product Inc.); Group 2, Activ GP (Brasseler); Group 3, SmartSeal (DFRP Ltd. Villa Farm); Group 4, AH 26 (Dentsply de Trey)/gutta-percha (GP). For push-out bond strength measurement, two horizontal slices were obtained from each specimen (n = 20). To compare dentinal tubule penetration, remaining 32 roots assigned to 4 groups as above were obturated with 0.1% Rhodamine B labeled sealers. One horizontal slice was obtained from the middle third of each specimen (n = 8) and scanned under confocal laser scanning electron microscope. Tubule penetration area, depth, and percentage were measured. Kruskall-Wallis test was used for statistical analysis. Results EndoRez showed significantly lower push-out bond strength than the others (p < 0.05). No significant difference was found amongst the groups in terms of percentage of sealer penetration. SmartSeal showed the least penetration than the others (p < 0.05). Conclusions The bond strength and sealer penetration of resin-and glass ionomer-based sealers used with coated core was not superior to resin-based sealer used with conventional GP. Dentinal tubule penetration has limited effect on bond strength. The use of conventional GP with sealer seems to be sufficient in terms of push-out bond strength.

Voltage-gated sodium channel genes and associated proteins have been cloned and studied in many mammalian and invertebrate species. However, there is no data available about the sodium channel gene(s) in the crayfish, although the animal has frequently been used as a model to investigate various aspects of neural cellular and circuit function. In the present work, by using RNA extracts from crayfish abdominal ganglia samples, the complete open reading frame of a putative sodium channel gene has firstly been cloned and molecular properties of the associated peptide have been analyzed. The open reading frame of the gene has a length of 5793 bp that encodes for the synthesis of a peptide, with 1930 amino acids, that is 82 % similar to the a-peptide of a sodium channel in a neighboring species, Cancer borealis. The transmembrane topology analysis of the crayfish peptide indicated a pattern of four folding domains with several transmembrane segments, as observed in other known voltage-gated sodium channels. Upon analysis of the obtained sequence, functional regions of the putative sodium channel responsible for the selectivity filter, inactivation gate, voltage sensor, and phosphorylation have been predicted. The expression level of the putative sodium channel gene, as defined by a qPCR method, was measured and found to be the highest in nervous tissue.