Search

Recently we have shown that crotamine, a toxin from the South American rattlesnake Crotalus durissus terrificus venom, belongs to the family of cell-penetrating peptides. Moreover, crotamine was demonstrated to be a marker of centrioles, of cell cycle, and of actively proliferating cells. Herein we show that this toxin at non-toxic concentrations is also capable of binding electrostatically to plasmid DNA forming DNA-peptide complexes whose stabilities overcome the need for chemical conjugation for carrying nucleic acids into cells. Interestingly, crotamine demonstrates cell specificity and targeted delivery of plasmid DNA into actively proliferating cells both in vitro and in vivo, which distinguishes crotamine from other known natural cell-penetrating peptides, The mechanism of crotamine penetration and cargo delivery into cells was also investigated, showing the involvement of heparan sulfate proteoglycans in the uptake phase, which is followed by endocytosis and peptide accumulation within the acidic endosomal vesicles. Finally, the permeabilization of endosomal membranes induced by crotamine results in the leakage of the vesicles contents to the cell cytosol. [ABSTRACT FROM AUTHOR]

this study evaluated dentin microtensile bond strength (µTBS) and failure modes (at 24 h and one year), bonding interface regarding hybridization, surface morphology regarding demineralization, in situ metalloproteinase (MMP) activity, and antibacterial effect of three dentin etchants compared to 35% phosphoric acid (PA). The Adper Single Bond 2 adhesive (3 M Oral Care) was applied on moist dentin etched with PA (control) or on air-dried dentin etched with 3% aluminum nitrate + 2% oxalic acid (AN), 6.8% ferric oxalate + 10% citric acid (FO), or 10% citric acid (CA). The µTBS test used 40 human teeth (n = 10). Failure modes and surface morphology were analyzed by scanning electron microscopy (n = 3), while bonding interface morphology and MMP activity were evaluated by laser scanning confocal microscopy (n = 3). Antibacterial activity was evaluated against S. Mutans biofilm by means of viable cells count (CFU/mL). PA presented the highest bond strengths regardless of aging time. PA, AN, and CA showed stable bond strengths after one year of storage. Adhesive and mixed failures were predominant in all groups. Thin hybrid layers with short resin tags were observed for the experimental etchants. The AN-based etchant was able to inhibit MMP activity. All tested etchants presented antibacterial activity against S. Mutans biofilm. This study suggests different dentin etchants capable of inhibiting MMP activity while also acting as cavity disinfectants. • Phosphoric acid bond strength was higher than the tested etchants. • Ferric oxalate + citric acid showed significant bond strength reduction at 1 year. • Gelatinolytic activity was inhibited by aluminum nitrate + oxalic acid. • Bacteria cell viability was reduced according to the pH of the acids. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the influence of surface treatment on roughness (SA), topography, and shear bond strength (SBS) of computer‐aided designer and manufacture (CAD/CAM) zirconia‐reinforced lithium silicate (ZLS) and feldspathic (FEL) glass–ceramics. Materials and methods: FEL and ZLS specimens were submitted to 5% or 10% hydrofluoric acid (HF) or self‐etching ceramic primer (MEP) and different application times (20, 40, and 60 s), or to sandblasting (Control, 20 s). Resin cement cylinders were bonded to the specimens and tested in shear (n = 10) after 24 h and 16‐months of water storage. SA and topography were evaluated by atomic force (AFM, n = 10) and scanning electron microscopy. Data were analyzed by ANOVA and Bonferroni test (α = 0.05). Results: Sandblasting promoted the highest SA for ZLS, but 10% HF (40, 60 s) promoted higher SBS at 16 months. 10% HF produced the highest SA for FEL, but sandblasting and 5% HF (20 s) maintained SBS after 16 months, without differences from 10% HF (20 s) (p > 0.05). Overall, MEP produced lower SA and SBS among groups (p < 0.05). HF displayed greater morphological changes on FEL. Conclusion: 10% HF (40 s) provided better results for ZLS, while 5% or 10% HF (20 s) was suitable for FEL. Clinical significance: Surface treatments influenced SA, topography, and SBS of materials. HF etching is the surface treatment of choice for both CAD/CAM glass–ceramics. [ABSTRACT FROM AUTHOR]

Purpose: Verify the influences of physical activity level, nutritional status and screen habits on the prevalence of back pain in Brazilian students. Methods: The sample consisted of 577 schoolchildren (female = 274; male = 303) aged between 10 and 16 years old, regularly enrolled in the 6th grade of elementary school living in the metropolitan area of the Alto Tietê of the state of São Paulo, Brazil. The prevalence, intensity and frequency of pain was verified with the Back Pain Assessment Instrument. The usual practice of physical activity was verified with the Physical Activity Questionnaire for Older Children/Adolescent. Nutritional status was analyzed using Body Mass Index. Screen habits were obtained through a previously structured questionnaire. Results: The Chi-square test indicated that pain complaint and its prevalence in the cervical region are significantly higher in females (p < 0.05). The multiple logistic regression test revealed that watching television influences the prevalence of cervical pain and that the use of more than one screen increases the occurrence of low back pain in male students (p < 0.05). Conclusion: Female students were the most affected by back pain complain, especially in the cervical region. However, factors associated with the prevalence of back pain were found only in males. [ABSTRACT FROM AUTHOR]

This study evaluated the cell viability and expression of different major genes involved in mineralization in odontoblast-like cells exposed to sodium trimetaphosphate (STMP). It was also investigated the influence of STMP on the rate of calcium phosphate crystal growth, its anti-proteolytic action against the enzymatic degradation of type I collagen, the binding mechanism of STMP to collagen fibrils, and the potential mechanism to induce collagen stabilization. Immortalized rat odontoblast MDPC-23 cells were cultured. Cell viability was assessed by trypan blue staining, and the changes in gene expression balance induced by STMP were assessed by quantitative reverse transcription (qRT) PCR assays. Crystalline particle formation was monitored by light-scattering detectors to estimate pH variation and the radial size of the crystalline particles as a function of reaction time (pH 7.4, 25 °C) in the presence of STMP in supersaturated calcium phosphate solution (Ca/P = 1.67). Images were obtained under atomic force microscopy (AFM) to measure the particle size in the presence of STMP. A three-point bending test was used to obtain the elastic modulus of fully demineralized dentin beams after immersion in STMP solution. The binding mechanism of STMP to collagen fibrils and potential stabilization mechanism was assessed with circular dichroism spectrometry (CD). The data were analyzed statistically (α = 0.05). STMP had no significant influence on the cell viability and gene expression of the MDPC-23 cells. STMP greatly increased the rate of crystal growth, significantly increasing the average radial crystal size. AFM corroborated the significant increase of STPM-treated crystal size. Mineralized collagen I fibrils exhibited less collagenase degradation with lower STMP concentration. CD analysis demonstrated changes in the conformational stability after STMP binding to type I collagen. The increased resistance of collagen against the proteolytic activity of collagenases appears to be related to the conformational change induced by STMP binding in collagen I and the STMP capacity for promoting biomimetic mineralization in type I collagen fibrils. [ABSTRACT FROM AUTHOR]

Abstract Objective Clinical issues have been raised about problems related to cytotoxic effects caused when applying self-adhesive cement. It was hypothesized that byproducts eluted from self-adhesive cements modulate oxidative stress response, the gene expression of signaling pathways of inflammatory process/transcriptional activators, and the expression and activity of interstitial collagenases, and modify the phenotypic characteristics of cellular proliferation and mineral deposition in odontoblastic-like cells. Methods Cements (MaxCem Elite [MAX] and RelyX U200 [U200)]) were mixed, dispensed into moulds, and photoactivated according to the manufacturers' instructions. Immortalized rat odontoblast-like cells (MDPC-23) were cultured and exposed to polymerized specimens of cements for 4 h. Reactive oxidative specimen production and quantification of gene expression were evaluated. Cell proliferation assay and alizarin red staining were also performed to evaluate the disturbance induced by the cements on cellular proliferation and mineralization. Results Despite their cytotoxic effects, both self-adhesive cements influenced the metabolism in the odontoblast cells on different scales. MAX induced significantly higher oxidative stress in odontoblast cells than U200. Gene expression varied as a function of exposure to self-adhesive cements; MAX induced the expression of pro-inflammatory cytokines such as TNF-α, whereas U200 downregulated, virtually depleted TNF-α expression, also inducing overexpression of the transcriptional factor Runx2. Overexpression of heme oxygenase-1 (HO-1) and thioredoxin reductase 1 (TRXR1) occurred after exposure to both cements, antioxidant genes that are downstream of Keap1-Nrf2-ARE system. MAX significantly induced the overexpression of collagenase MMP-1, and U200 induced the expression of gelatinase MMP-2. MAX significantly inhibited cell proliferation whereas U200 significantly activated cell proliferation. Alizarin red staining revealed significantly decreased mineral deposition especially when exposed to MAX. Significance These results support the hypothesis that byproducts of different self-adhesive cements play important roles in the highly orchestrated process which ultimately affect the cellular proliferation and the mineral deposition in odontoblastic-like cells, possibly delaying the reparative dentin formation after cementation of indirect restorations, especially on recently exposed dentin preparations. [ABSTRACT FROM AUTHOR]

Objectives To evaluate the cytotoxic effects of exposing odontoblast cells to a variety of commercial self-adhesive cements polymerized using different activation modes. Methods Five cements: MaxCem Elite (MAX), Bifix SE (BSE), G-Cem LinkAce (GCE), Clearfil SA Luting (CAS), and RelyX U200 (U200) were mixed, dispensed into molds, and distributed in groups, according to polymerization protocols: immediate photoactivation; delayed photoactivation (10 min self-curing plus light-activation); and chemical activation (no light exposure). Immortalized rat odontoblast cells (MDPC-23) were cultured. Cell viability was assessed by Trypan Blue staining and total cell death was assessed by annexin V-APC/7-AAD double staining and flow cytometry. Volatilized compounds from polymerized specimens of cements were evaluated by gas chromatography/mass spectrometry (GC–MS). Data was analyzed with 2-way ANOVA/Tukey tests ( α = 0.05). Results Exposure to all of the cements tested significantly reduced the cell viability, irrespective of the activation protocol (p < 0.05). The least harmful cements were CSA and U200. Total death of cells significantly increased when exposed to BSE, GCE, and MAX, especially when chemically activated (p < 0.05). Characteristic apoptotic cells increased after exposure to cements, mainly for MAX, regardless of the activation mode. Chemical activation of MAX also induced necrosis. Moreover, GCE and MAX exhibited higher percentages of late apoptotic/dead cells. Chromatograms revealed 28 compounds released from the cements tested, some of them with known carcinogenic effects. Selection of self-adhesive cements and polymerization protocols affect the cytotoxicity and cell viability of odontoblastic cells. Clinical significance Despite the simplified cementation protocol, care is needed when cementing indirect restorations with self-adhesive cements, especially on recently exposed dentin. This category of material may cause differential cytotoxic effects and should be considered when selecting a cement. This is particularly true in clinical cases of light attenuation, where the polymerization depends on chemical activation, inducing higher cytotoxic damages when using some of the cements tested. [ABSTRACT FROM AUTHOR]

This study evaluated dentin bond strength and dentin-resin interfacial morphology using phosphoric acid-etching and EDTA-conditioning. The occlusal enamel of twenty-four human third molars was removed to expose the dentin surfaces, which were polished with a SiC paper (600 grit). Teeth were randomly divided into three groups (n = 8): etching with 37% phosphoric acid (15 s), conditioning with 0.1 M EDTA (60 s) and 0.5 M EDTA (120 s). Adhesive (XP Bond, Dentsply) was applied and three layers (2 mm each) of composite (EsthetX, Dentsply) were placed and light-activated separately (20 s). Teeth were sectioned to obtain specimens for the microtensile bond strength test (1 mm 2 at cross section). Half of the specimens was tested in a universal testing machine (EZ Test, Shimadzu) after 24 h and the other half after storage for 10 months. Failure mode and adhesive interface were analyzed using scanning electron microscopy. Data were analyzed by mixed models for repeated measures (PROC MIXED) and Tukey-Kramer test, considering a significance level of 5%. Mean bond strength values (SD) after 24 h and 10 months were, respectively (in MPa): phosphoric acid: 37.3 (7.7) / 33.9 (6.7); 0.1 M EDTA: 14.7 (7.3) / 15.1 (10.1); 0.5 M EDTA: 25.1 (7.7) / 21.1 (14.1). Dentin treatment with EDTA and phosphoric acid resulted in hybrid layer and resin tags formation. Mixed failures were prevalent for all groups tested after 24 h. Storage for 10 months changed the failure mode for the 0.5 M EDTA group to adhesive failure (between dentin and bonding agent). EDTA applied for 60 s yielded lower bond strength results compared to phosphoric acid and EDTA (120 s), but there was no significant reduction after 10 months of storage for any of the tested groups. [ABSTRACT FROM AUTHOR]

It has been hypothesized that cysteine cathepsins (CTs) along with matrix metalloproteases (MMPs) may work in conjunction in the proteolysis of mature dentin matrix. The aim of this study was to verify simultaneously the distribution and presence of cathepsins B (CT-B) and K (CT-K) in partially demineralized dentin; and further to evaluate the activity of CTs and MMPs in the same tissue. The distribution of CT-B and CT-K in sound human dentin was assessed by immunohistochemistry. A double-immunolabeling technique was used to identify, at once, the occurrence of those enzymes in dentin. Activities of CTs and MMPs in dentin extracts were evaluated spectrofluorometrically. In addition, in situ gelatinolytic activity of dentin was assayed by zymography. The results revealed the distribution of CT-B and CT-K along the dentin organic matrix and also indicated co-occurrence of MMPs and CTs in that tissue. The enzyme kinetics studies showed proteolytic activity in dentin extracts for both classes of proteases. Furthermore, it was observed that, at least for sound human dentin matrices, the activity of MMPs seems to be predominant over the CTs one. [ABSTRACT FROM AUTHOR]