Antimicrobial and optical properties of a new biogenic silica-coated silver nanoparticles incorporated into experimental resin.

Background: Evaluate the effects of incorporating silica-coated silver nanoparticles (Ag@SiO2 NPs) into odontological clinic resin materials. Material and Methods: Silver nanoparticles coated with silicon dioxide were added to the experimental resin matrix at 1, 3, and 5wt%. Degree of conversion (DC), optical properties (total transmittance and color change), and microstructural analysis were evaluated. Materials were tested for silver ion release, cytotoxicity in dental pulp fibroblasts, Streptococcus mutans biofilm growth by Colony-Forming Unit (CFU) and confocal laser scanning microscopy (CLSM). Results: Groups had a similar DC, despite significant differences observed in transmittance and color change analysis for all groups with NPs. Silver ion release values were below the detection limit after 72h for all groups, and NPs incorporation did not show a statistical difference from the control on pulp fibroblasts assay. After 72h, the CFU count was significantly reduced by 74% from 3wt% of Ag@SiO2NPs. CLSM evaluation on S. mutans colonies showed a dose-dependent decrease in the emitted fluorescence. Conclusions: The application of Ag@SiO2 NPs in a resinous matrix, demonstrates a significant reduction of S. mutans CFU in oral biofilm, at concentrations from 3wt%, without an increase in cytotoxicity. The reduced transmittance values did not affect the DC, although a significant color change was perceived in all concentrations. Key words:Nanoparticles, Silver Compounds, Composite Dental Resin, Anti-Bacterial Agent, Optical Imaging.

Effect of experimental dentin etchants on dentin bond strength, metalloproteinase inhibition, and antibiofilm activity.

OBJECTIVE: 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). MATERIALS AND METHODS: 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). RESULTS: 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. SIGNIFICANCE: This study suggests different dentin etchants capable of inhibiting MMP activity while also acting as cavity disinfectants.

Poly(Aspartic Acid) Promotes Odontoblast-like Cell Differentiation in Rat Molars with Exposed Pulp.

In recent years, alternative pulpal therapies targeting dentinogenesis signaling pathways using different peptides have been investigated. The aim of this study was to verify the effectiveness of poly(aspartic acid), pAsp, in dentin regeneration using an animal model. METHODS: Mechanical pulp exposure was performed in the upper molars of 56 Wistar rats, randomly divided as follows (n = 14): control (no treatment); MTA group-pulp capping with mineral trioxide aggregate (MTA Angelus); pAsp group-application of 20 µL of pAsp solution (25 mg·mL-1); MTA+pAsp group-application of MTA mixed with pAsp (5:1 by mass). Animals were euthanized after 7 or 21 days. Histological sections were submitted to hematoxylin-eosin and Brown and Brenn staining and immunohistochemical analysis for osteopontin (OPN) and dentin matrix protein 1 (DMP 1). RESULTS: At 7 days, an acute inflammatory infiltrate and the presence of disorganized mineralized tissue were observed in all groups. At 21 days, the quality and thickness of the reparative dentin in treated groups were superior to the control, and bacterial contamination was observed in two MTA-pAsp specimens. While all treated groups showed intense immunostaining for OPN at 21 days, only the pAsp group expressed DMP 1, indicating the presence of fully differentiated odontoblast-like cells. CONCLUSION: Poly(aspartic) acid promoted dentin regeneration in rat molars in the absence of an additional calcium source and may be an alternative to MTA as a pulp-capping agent.

Self-assembled peptide P11-4 interacts with the type I collagen C-terminal telopeptide domain and calcium ions.

OBJECTIVES: Evaluate molecularly the role of P11-4 self-assembly peptide in dentin remineralization and its interaction with collagen I. METHODS: The calcium-responsive P11-4 peptide was analyzed by intrinsic fluorescence emission spectrum, circular dichroism spectrum (CD), and atomic force microscope (AFM). Differential light scattering was used to monitor the nucleation growth rate of calcium phosphate nanocrystals in the absence or in the presence of P11-4. AFM was used to analyze the radial size (nm) of calcium phosphate nanocrystals formed in the absence or in the presence of P11-4, as well as to verify the spatial structure of P11-4 in the absence or in the presence of Ca2+. RESULTS: The interaction of Ca2+ with the P11-4 (KD = 0.58 ± 0.06 mM) promotes the formation of ß-sheet antiparallel structure, leads to its precipitation in saturated solutions of Ca/P = 1.67 and induces the formation of parallel large fibrils (0.6 - 1.5 µm). P11-4 organized the HAP nucleation by reducing both the growth rate and size variability of nanocrystals, analyzed by the F test (p < 0.0001, N = 30). P11-4 interacts (KD = 0.75 ± 0.06 µM) with the KGHRGFSGL motif present at the C-terminal collagen telopeptide domain. P11-4 also increased the amount of HAP and collagen in the MDPC-23 cells. SIGNIFICANCE: The presented data propose a mechanism that will help future clinical and/or basic research to better understand a molecule able to inhibit structural collagen loss and help the impaired tissue to remineralize.

Effects of argon plasma and aging on the mechanical properties and phase transformation of 3Y-TZP zirconia.

To evaluate the flexural strength (FS) and flexural modulus (FM) of a commercial 3Y-TZ0P ceramic after artificial aging and either without or with two application times of non-thermal plasma treatments (NTP). In addition, changes in crystalline phase transformation and surface nano-topography after NTP application, during different aging periods, were evaluated. Ninety 3Y-TZP bars (45x4x3 mm) were made for FS and FM testing, and assigned to nine groups (n=10): no NTP/no aging (Control); no NTP/4h aging; no NTP/30h aging; 10s NTP/no aging; 10s NTP/4h aging; 10s NTP/30h aging; 60s NTP/no aging; 60s NTP/4h aging and 60s NTP/30h aging. Artificial accelerated aging was simulated using an autoclave (134º C at 2 bar) for up to 30h. FS and FM were assessed using a universal testing machine and data analyzed using a ANOVA and Tukey test (α=0.05). The volume change in zirconia monoclinic phase (MPV) was evaluated using X-ray diffraction and surface nano-topography was assessed using atomic force microscopy (baseline until 30h-aging). NTP application did not influence the FS and FM of zirconia. Compared to the Control (no NTP/no aging), the FS of zirconia samples treated for 30 hours in autoclave ("no NTP/30h aging" group) increased. Artificial aging for 30 hours significantly increased the FM of zirconia, regardless of NTP application. MPV tended to increase following the increase in aging time, which might result in the surface irregularities observed at 30h-aging. NTP did not alter the zirconia properties tested, but 30h-aging can change the zirconia FS, FM and MPV.

Effects of argon plasma and aging on the mechanical properties and phase transformation of 3Y-TZP zirconia

Abstract To evaluate the flexural strength (FS) and flexural modulus (FM) of a commercial 3Y-TZ0P ceramic after artificial aging and either without or with two application times of non-thermal plasma treatments (NTP). In addition, changes in crystalline phase transformation and surface nano-topography after NTP application, during different aging periods, were evaluated. Ninety 3Y-TZP bars (45x4x3 mm) were made for FS and FM testing, and assigned to nine groups (n=10): no NTP/no aging (Control); no NTP/4h aging; no NTP/30h aging; 10s NTP/no aging; 10s NTP/4h aging; 10s NTP/30h aging; 60s NTP/no aging; 60s NTP/4h aging and 60s NTP/30h aging. Artificial accelerated aging was simulated using an autoclave (134º C at 2 bar) for up to 30h. FS and FM were assessed using a universal testing machine and data analyzed using a ANOVA and Tukey test (α=0.05). The volume change in zirconia monoclinic phase (MPV) was evaluated using X-ray diffraction and surface nano-topography was assessed using atomic force microscopy (baseline until 30h-aging). NTP application did not influence the FS and FM of zirconia. Compared to the Control (no NTP/no aging), the FS of zirconia samples treated for 30 hours in autoclave ("no NTP/30h aging" group) increased. Artificial aging for 30 hours significantly increased the FM of zirconia, regardless of NTP application. MPV tended to increase following the increase in aging time, which might result in the surface irregularities observed at 30h-aging. NTP did not alter the zirconia properties tested, but 30h-aging can change the zirconia FS, FM and MPV.

Resumo Avaliar a resistência à flexão (FS) e o módulo de flexão (FM) de uma cerâmica comercial 3Y-TZP após envelhecimento artificial, e com ou sem dois tempos de aplicação de plasma não térmico (NTP). Além disso, a transformação de fase cristalina e a nano-topografia de superfície após a aplicação de NTP, durante diferentes períodos de envelhecimento, foram avaliadas. Noventa barras 3Y-TZP (45x4x3 mm) foram feitas para testes de FS e FM, e distribuídas em nove grupos (n=10): sem NTP/sem envelhecimento (Controle); sem NTP/4h de envelhecimento; sem NTP/30h de envelhecimento; 10sNTP/sem envelhecimento; 10sNTP/4h; 10sNTP/30h; 60sNTP/sem envelhecimento; 60sNTP/4h e 60sNTP/30h. O envelhecimento artificial acelerado foi simulado em autoclave (134º C a 2 bar) por até 30 horas. FS e FM foram avaliados em máquina de ensaio universal e os dados analisados ​​pela ANOVA e teste de Tukey (α = 0,05). A mudança de volume da fase monoclínica de zircônia (MPV) foi avaliada usando difração de raios-X e nano-topografia de superfície foi avaliada utilizando microscopia de força atômica (baseline até 30h). A aplicação do NTP não influenciou a FS e FM da zircônia. Comparado ao Controle ("sem NTP/sem envelhecimento"), a FS das amostras de zircônia tratadas por 30 horas em autoclave ("sem NTP/30h de envelhecimento") aumentou. O envelhecimento artificial por 30 horas aumentou significativamente a FM da zircônia, independente do tempo de aplicação do NTP. O MPV tendeu a aumentar em função do aumento do tempo de envelhecimento para todos os grupos, que pode ter resultado nas irregularidades superficiais observadas com 30 horas de envelhecimento. O NTP não afetou as propriedades da zircônia testadas, mas o envelhecimento por 30 horas pode alterar a FS, FM e MPV da zircônia.

Surface treatments on CAD/CAM glass-ceramics: Influence on roughness, topography, and bond strength.

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.

Back pain in elementary schoolchildren is related to screen habits.

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.

Photobiomodulation Therapy Modulates Muscle Gene Expression and Improves Performance of Rats Subjected to a Chronic Resistance Exercise Protocol.

Objective: In professional sports activities, the search for increased performance is constant. Electrophysical agents, including photobiomodulation (PBM), have been used in the sports context to accelerate postworkout recovery, prevent injuries, and even to improve performance. This study aims to investigate the effects of infrared laser (904 nm) on skeletal muscle gene expression of performance-related proteins of rats submitted to a chronic resistance training protocol. Materials and methods: Male Wistar rats (n = 40), weighing ±300 g were divided into four groups: sedentary control (CT, n = 10); irradiated control (CTL, n = 10); exercised not irradiated (EX, n = 10); exercised irradiated (EXL, n = 10). To assess the performance, the maximum carrying test was adapted and applied 72 h prior the training and 72 h after the last exercise session. The vertical weight climbing protocol was adapted for resistance training 3 × per week with 48 h interval between each session: first week adaptation, second week 25% of body weight (BW), third week 50% BW, fourth week 75% BW, and fifth week 100% BW. Animals were irradiated before exercise on hind paws 50 sec each, with infrared laser 904 nm 5 days per week, during 4 weeks, 9 J per leg in a total of 18 J energy per day. Results: The EXL performed more climbing (7.1 ± 0.91) compared to EX (4.4 ± 0.63). PBM promoted increased expression of lactate dehydrogenase enzyme, mammalian target of rapamycin protein, and androgen receptor (p < 0.05) but not the myosin heavy chain (p = 0.43). Conclusions: PBM therapy increases the expression of performance-related muscle mass gain genes besides improving the resistance training performance.

The interaction of sodium trimetaphosphate with collagen I induces conformational change and mineralization that prevents collagenase proteolytic attack.

OBJECTIVES: 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. METHODS: 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). RESULTS: 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. SIGNIFICANCE: 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.

904 nm Low-Level Laser Irradiation Decreases Expression of Catabolism-Related Genes in White Adipose Tissue of Wistar Rats: Possible Roles of Laser on Metabolism.

Background: Adipose tissue is the main energy storage tissue in the body. Its catabolic and anabolic responses depend on several factors, such as nutritional status, metabolic profile, and hormonal signaling. There are few studies addressing the effects of laser photobiomodulation (PBM) on adipose tissue and results are controversial. Objective: Our purpose was to investigate the metabolic effects of PBM on adipose tissue from Wistar rats supplemented or not with caffeine. Materials and methods: Wistar rats were divided into four groups: control (CTL), laser-treated [CTL (L)], caffeine (CAF), and caffeine+PBM [CAF (L)]. Blood was extracted for quantification of triglyceride and cholesterol levels and white adipose tissues were collected for analysis. We evaluated gene expression in the adipose tissue for the leptin receptor, lipase-sensitive hormone, tumor necrosis factor alpha, and beta adrenergic receptor. Results: We demonstrated that the low-level laser irradiation was able to increase the feed intake of the animals and the relative mass of the adipose tissue in the CTL (L) group compared with CTL. Laser treatment also increases serum triglycerides [CTL = 46.99 ± 5.87; CTL (L) = 57.46 ± 14.38; CAF = 43.98 ± 5.17; and CAF (L) = 56.9 ± 6.12; p = 0.007] and total cholesterol (CTL = 70.62 ± 6.80; CTL (L) = 79.41 ± 13.07; CAF = 71.01 ± 5.52; and CAF (L) = 79.23 ± 6.881; p = 0.003). Conclusions: Laser PBM decreased gene expression of the studied genes in the adipose tissue, indicating that PBM is able to block the catabolic responses of this tissue. Interestingly, the CAF (L) and CAF animals presented the same CLT (L) phenotype, however, without increasing the feed intake and the relative weight of the adipose tissue. The description of these phenomena opens a new perspective for the study of the action of low-level laser in adipose tissue.

Laser Photobiomodulation 904 nm Promotes Inhibition of Hormone-Sensitive Lipase Activity in 3T3-L1 Adipocytes Differentiated Cells.

Background: The lipid metabolism is essential for maintaining the body's energy responses. Laser photobiomodulation triggers many important cellular effects, but these effects on lipid metabolism are not well described. In this study, we analyzed the laser photobiomodulation in the hormone-sensitive lipase (HSL) activity, a key enzyme in the triglycerides (TAG) hydrolysis in adipose tissue 3T3-L1. Methods: Cells were submitted to the differentiation protocol in adipose cells, irradiated with 1, 2, and 3J with laser (904 nm-60 mw-laser diode) and incubated for 4 h after irradiation. Results: The response of laser photobiomodulation was able to trigger an inhibition of HSL activity (control = 0.057 ± 0.0008; 1J = 0.050 ± 0.0003; 2J = 0.0477 ± 0.002; 3J = 0.051 ± 0.002; p = 0.0003 against the control), but no modulation was observed in TAG levels into the medium (control = 26.5856 ± 0.52; 1J = 26.5856 ± 0.52; 2J = 27.2372 ± 1.41; 3J = 25.9991 ± 0.1303; p = 0.18). Conclusions: This is the first study of HSL activity modulation with laser radiation, suggesting that photobiomodulation can influence adipose tissue metabolism and open a new field of study.

Oxidative stress induced by self-adhesive resin cements affects gene expression, cellular proliferation and mineralization potential of the MDPC-23 odontoblast-like cells.

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.

Differential cytotoxic effects on odontoblastic cells induced by self-adhesive resin cements as a function of the activation protocol.

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 (10min 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.

Mini-interfacial Fracture Toughness of a Multimode Adhesive Bonded to Plasma-treated Dentin.

PURPOSE: To investigate the bonding efficacy of a multimode adhesive to plasma-treated and -untreated (control) dentin using a mini-interfacial fracture toughness (mini-iFT) test. MATERIALS AND METHODS: Twenty human molars were used in a split-tooth design (n = 10). The adhesive Scotchbond Universal (SBU; 3M ESPE) was applied in etch-and-rinse (E&R) and self-etch (SE) modes. Mid-coronal dentin was exposed and covered with a standardized smear layer ground to 320 grit. One half of each dentin surface received 15 s of non-thermal atmospheric plasma (NTAP), while the other half was covered with a metallic barrier and kept untreated. Following the E&R mode, dentin was plasma treated immediately after phosphoric acid etching. SBU and a resin-based composite were applied to dentin following the manufacturer's instructions. Six mini-iFT specimens were prepared per tooth (1.5 x 2.0 x 16 to 18 mm), and a single notch was prepared at the adhesive-dentin interface using a 150-µm diamond blade under water cooling. Half of the mini-iFT specimens were immediately loaded until failure in a 4-point bending test, while the other half were first stored in distilled water for 6 months. After testing, the exact dimensions of the notch were measured with a measuring optical microscope, from which ΚIc was determined. RESULTS: Three-way ANOVA revealed higher mini-iFT for SBU applied in E&R than SE mode for both storage times, irrespective of NTAP treatment. CONCLUSION: Overall, mini-iFT did not decrease for any of the experimental groups upon 6-month aging, while plasma treatment did not show a direct beneficial effect on mini-iFT of SBU applied in either E&R or SE mode.

Co-distribution of cysteine cathepsins and matrix metalloproteases in human dentin.

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.

Crotamine mediates gene delivery into cells through the binding to heparan sulfate proteoglycans.

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.