Measurement of four-photon absorption in GaP and ZnTe semiconductors.

Intensity-dependent effective four-photon absorption (4PA) coefficients in GaP and ZnTe semiconductors were measured by the z-scan method using pump pulses of 1.75 µm wavelength, 135 fs duration, and up to 500 GWcm-2 intensity. A nonlinear pulse propagation model, including linear dispersion and 4PA was used to obtain the 4PA coefficients from measurements. The intensity-dependent effective 4PA coefficients vary from 2.6 × 10-4 to 65 × 10-4 cm5GW-3 in GaP, and from 3.5 × 10-4 to 9.1 × 10-4 cm5GW-3 in ZnTe. The anisotropy in 4PA was shown in GaP. The knowledge of 4PA coefficients is important for the design of semiconductor photonics devices.

Transdentinal photobiostimulation of stem cells from human exfoliated primary teeth.

AIM: To evaluate the effects of infrared light-emitting diode (LED) irradiation on stem cells from human exfoliated deciduous teeth (SHEDs). METHODOLOGY: Exfoliated primary teeth were obtained (n = 3), and SHEDs obtained from the teeth were seeded on the pulpal surface of 0.2-mm-thick dentine discs produced from permanent molars. The cells were incubated for 24 h by placing the discs in plain Dulbecco's modified Eagle's medium (DMEM). The DMEM was then replaced with new culture medium formulated for odontoblast differentiation. After 12 h in the second medium, SHEDs were irradiated through the dentine discs using an infrared LED (850 nm) with a power density of 80 mW cm-2 . Energy doses (EDs) delivered to the occlusal surface of the dentine discs were 0 (control), 2 and 4 J cm-2 (n = 6). Subsequent tests were performed 72 h after irradiation. These tests included cell viability (MTT), alkaline phosphatase activity (ALP), total protein production (TP), scanning electron microscopy (SEM), as well as gene expression for ALP, Col I, DSPP and DMP-1. Data were analysed using Kruskal-Wallis and Mann-Whitney t-tests (α = 0.05). RESULTS: Both EDs (2 and 4 J cm-2 ) significantly increased cell viability and ALP activity. For TP, ALP and Col I gene expression, only the 4 J cm-2 group had significantly higher values compared to the control group. Cell morphology was not affected by irradiation. CONCLUSION: Infrared LED irradiation was capable of biostimulating SHEDs through a 0.2 mm thickness of dentine, especially at the 4 J cm-2 level.

Influence of enamel/dentin thickness on the toxic and esthetic effects of experimental in-office bleaching protocols.

OBJECTIVES: This paper aims to assess the whitening effectiveness and toxicity of tooth-bleaching protocols applied to enamel/dentin disks simulating mandibular incisors (ICs) and premolars (PMs). MATERIALS AND METHODS: A 10% hydrogen peroxide (H2O2) gel was applied for 3 × 15, 1 × 15, or 1 × 5 min to enamel/dentin disks simulating mandibular ICs and PMs, and the trans-enamel and trans-dentinal diffusion products were applied to human dental pulp cells (1 h). Professional therapy (35% H2O2-3 × 15 min) was used as positive control, and non-bleached samples were used as negative control. Cell viability and morphology, oxidative stress generation, and odontoblastic marker expression were assessed. The H2O2 diffusion and enamel color change (ΔE) were also analyzed. RESULTS: The 10% H2O2 gel induced significant cell viability reduction only when applied 3 × 15 min, with the intensity of oxidative stress and down-regulation of odontoblastic markers being higher in the IC group. The other experimental bleaching protocols caused slight alterations regarding the cell parameters evaluated, with intensity being related to enamel/dentin thickness. These effects were also correlated with higher H2O2 diffusion in the IC group. ΔE values similar as positive control were found for the 10% 3 × 15 and 1 × 15 protocols on IC group, after 4 and 6 sessions. CONCLUSION: Application of a 10% H2O2 bleaching gel for 15 or 45 min to thin dental substrate significantly minimizes cell toxicity in comparison with highly concentrated gels associated with similar esthetic outcomes by increasing the number of bleaching sessions. CLINICAL RELEVANCE: Bleaching gels with 10% H2O2 applied in small teeth for short periods may be an interesting alternative to obtain whitening effectiveness without causing toxicity to pulp cells, which may be able to reduce the tooth hypersensitivity claimed by patients.

Hybrid tilted-pulse-front excitation scheme for efficient generation of high-energy terahertz pulses.

Conception of a hybrid type tilted-pulse-front pumping scheme for the generation of high-energy terahertz pulses is presented. The proposed setup is the combination of the conventional setup containing imaging optics and the contact grating. The solution was developed for nonlinear materials requiring large pulse-front-tilt angle, like LiNbO3. Due to the creation of the pulse-front-tilt in two steps the limitations of imaging errors can be significantly reduced. Furthermore the necessary grating constant of the contact grating can be larger compared to the simple contact grating scheme making possible the fabrication of the grating profile with significantly higher precision. A detailed optimization procedure with respect to the diffraction efficiency on the contact grating is given for LiNbO3. Instructions are also given how to construct the geometry of the setup in order to minimize imaging errors. Examples are given for LiNbO3 based practically realizable, optimized schemes with reduced imaging errors and high diffraction efficiency on the contact grating.

High-energy terahertz pulses from semiconductors pumped beyond the three-photon absorption edge.

A new route to efficient generation of THz pulses with high-energy was demonstrated using semiconductor materials pumped at an infrared wavelength sufficiently long to suppress both two- and three-photon absorption and associated free-carrier absorption at THz frequencies. For pumping beyond the three-photon absorption edge, the THz generation efficiency for optical rectification of femtosecond laser pulses with tilted intensity front in ZnTe was shown to increase 3.5 times, as compared to pumping below the absorption edge. The four-photon absorption coefficient of ZnTe was estimated to be ß4=(4±1)×10-5 cm5/GW3. THz pulses with 14 µJ energy were generated with as high as 0.7% efficiency in ZnTe pumped at 1.7 µm. It is shown that scaling the THz pulse energy to the mJ level by increasing the pump spot size and pump pulse energy is feasible.

Indirect cytocompatibility of a low-concentration hydrogen peroxide bleaching gel to odontoblast-like cells.

AIM: To assess the initial cytotoxicity and the late phenotype marker expression of odontoblast-like cells (MDPC-23) subjected to less aggressive in-office bleaching therapies. METHODOLOGY: A 17.5% hydrogen peroxide (H2O2) gel was applied for 45, 15 or 5 min to enamel/dentine discs adapted to trans-wells positioned over cultured MDPC-23 cells. No treatment was performed on the negative control. Immediately after bleaching, the cell viability, gene expression of inflammatory mediators and quantification of H2O2 diffusion were evaluated. The ALP activity, DSPP and DMP-1 gene expression and mineralized nodule deposition (MND) were assessed at 7, 14 or 21 days post-bleaching and analysed statistically with Mann-Whitney U-tests (α = 5%). RESULTS: H2O2 diffusion, proportional to treatment time, was observed in all bleached groups. Reductions of approximately 31%, 21% and 13% in cell viability were observed for the 45-, 15- and 5-min groups, respectively. This reduction was significant (P < 0.05) for the 45- and 15-min groups, which also presented significant (P < 0.05) over-expression of inflammatory mediators. The 45-min group was associated with significant (P < 0.05) reductions in DMP-1/DSPP expression at all periods, relative to control. The ALP activity and MND were reduced only in initial periods. The 15-min group had less intense reduction of all markers, with no difference to control at 21 days. CONCLUSIONS: The 17.5% H2O2 applied to tooth specimens for 5 min caused no alteration in the odontoblast-like cells. When this gel was applied for 45 or 15 min, a slight cytotoxicity, associated with alterations in phenotypic markers, was observed. However, cells were able to recover their functions up to 21 days post-bleaching.

Cytocompatibility of HEMA-free resin-based luting cements according to application protocols on dentine surfaces.

AIM: To evaluate the transdentinal cytotoxicity of resin-based luting cements (RBLCs), with no HEMA in their composition, to odontoblast-like cells. METHODOLOGY: Human dentine discs 0.3 mm thick were adapted to artificial pulp chambers (APCs) and placed in wells of 24-well plates containing 1 mL of culture medium (DMEM). Two categories of HEMA-free RBLCs were evaluated: group 1, self-adhesive Rely X Unicem (RU; 3M ESPE), applied directly to the dentine substrate; and group 2, Rely X ARC (RARC; 3M ESPE), applied to dentine previously acid-etched and treated with a bonding agent. In group 3 (control), considered as representing 100% cell metabolic activity, no treatment was performed on dentine. The APC/disc sets were incubated for 24 h or 7 days at 37 °C and 5% CO2 . Then, the extracts (DMEM + dental materials components that diffused through dentine) were applied to cultured odontoblast-like MDPC-23 cells for 24 h. After that, the cell viability (MTT assay), cell morphology (SEM), total protein production (TP) and alkaline phosphatase (ALP) activity were assessed. Data from MTT assay and TP production were analysed by Kruskal-Wallis and Mann-Whitney tests (α = 5%). Data from ALP activity were analysed by one-way anova and Tukey's test (α = 5%). RESULTS: In group 1, a slight reduction in cell viability (11.6% and 16.8% for 24-h and 7-day periods, respectively) and ALP activity (13.5% and 17.9% for 24-h and 7-day periods, respectively) was observed, with no significant difference from group 3 (control) (P > 0.05). In group 2, a significant reduction in cell viability, TP production and ALP activity compared with group 3 (control) occurred (P < 0.05), regardless of incubation time. Alteration in MDPC-23 cell morphology was observed only in group 2. CONCLUSIONS: HEMA-free Rely X ARC cement caused greater toxicity to odontoblast-like MDPC-23 cells than did Rely X Unicem cement when both resin-based luting materials were applied to dentine as recommended by the manufacturer.

Circularly polarized carrier-envelope-phase stable attosecond pulse generation based on coherent undulator radiation.

In this Letter, we present a new method for generation of circularly polarized attosecond pulses. According to our calculations, shape-controlled, carrier-envelope-phase stable pulses of several hundred nanojoule energy could be produced by exploitation of the coherent undulator radiation of an electron bunch. Our calculations are based on an existing particle accelerator system (FLASH II in DESY, Germany). We investigated the energy dependence of the attosecond pulses on the energy of electrons and the parameters of the radiator undulator, which generate the electromagnetic radiation.

Efficient generation of THz pulses with 0.4 mJ energy.

Efficient generation of THz pulses with high energy was demonstrated by optical rectification of 785-fs laser pulses in lithium niobate using tilted-pulse-front pumping. The enhancement of conversion efficiency by a factor of 2.4 to 2.7 was demonstrated up to 186 µJ THz energy by cryogenic cooling of the generating crystal and using up to 18.5 mJ/cm2 pump fluence. Generation of THz pulses with more than 0.4 mJ energy and 0.77% efficiency was demonstrated even at room temperature by increasing the pump fluence to 186 mJ/cm2. The spectral peak is at about 0.2 THz, suitable for charged-particle manipulation.

Proposal for carrier-envelope-phase stable single-cycle attosecond pulse generation in the extreme-ultraviolet range.

A robust method for producing half-cycle-few-cycle attosecond pulses in the extreme ultraviolet spectral range is proposed. It is based on coherent undulator radiation of relativistic ultrathin electron layers (nanobunches), which are produced by nanobunching of ultrashort electron bunches by a 10-TW power laser in a modulator undulator. Our numerical calculations predict the generation of nanobunches shorter than 10 nm. By using these electron nanobunches the production of carrier-envelope-phase stable attosecond pulses with up to a few tens of nJ energy and down to 10 nm wavelength and 35 as duration is predicted.

Infrared LED irradiation photobiomodulation of oxidative stress in human dental pulp cells.

AIM: To investigate the effect of infrared light-emitting diode (LED) irradiation on the oxidative stress induced in human dental pulp cells (HDPCs) by lipopolysaccharide (LPS). METHODOLOGY: Human dental pulp cells (HDPCs) were harvested from sound primary teeth that were near exfoliation. Cells were seeded (10(5)  cells cm(-2) ) using α-MEM supplemented with 10% FBS and after 24 h, were placed in contact with LPS (10 µg mL(-1) of culture medium). Immediately afterwards, HDPCs were subjected to a single irradiation with an infrared LED (855 nm) delivering different doses of energy (0, 2, 4, 8, 15 or 30 J cm(-2) ). For each dose, there was a control group without LPS application. Twenty-four hours after irradiation, groups were tested for nitric oxide (NO) quantification, cell viability (MTT assay) and qualitative assessment of reactive oxygen species (ROS). Data were submitted to Kruskal-Wallis and Mann-Whitney tests (α = 0.05). RESULTS: Lipopolysaccharide (LPS)-induced stress resulted in significant increase in NO production by HDPC without causing damage to cell respiratory metabolism. Irrespective of energy dose delivered, NO production was significantly reduced when LPS-stressed cells were irradiated with infrared LED (2 J cm(-2) , P = 0.003; 95% CI = 5.84-27.71; 4 J cm(-2) , P = 0.001; 95% CI = 7.52-26.39; 8 J cm(-2) , P = 0.0195; 95% CI = -2.86-16.01; 15 J cm(-2) , P = 0.0001; 95% CI = 12.10-30.96; 30 J cm(-2) , P = 0.007; 95% CI = 5.84-24.71). The highest decrease in NO production was observed when 15 J cm(-2) was delivered to cells. Infrared LED irradiation resulted in a decrease in ROS production, whilst HDPC metabolism was not significantly affected. CONCLUSION: Biomodulation of oxidative stress of HPDC can be achieved by irradiation with a single dose of infrared LED. Within the range investigated, 15 J cm(-2) resulted in the least production of NO.

Generation of sub-mJ terahertz pulses by optical rectification.

Recent theoretical calculations predicted an order-of-magnitude increase in the efficiency of terahertz pulse generation by optical rectification in lithium niobate when 500 fs long pump pulses are used, rather than the commonly used ~100 fs pulses. Even by using longer than optimal pump pulses of 1.3 ps duration, 2.5× higher THz pulse energy (125 µJ) was measured with 2.5× higher pump-to-THz energy conversion efficiency (0.25%) than reported previously with shorter pulses. These results verify the advantage of longer pump pulses and support the expectation that mJ-level THz pulses will be available by cooling the crystal and using large pumped area.

Chitosan in association with osteogenic factors as a cell-homing platform for dentin regeneration: Analysis in a pulp-in-a-chip model.

OBJECTIVE: In this paper we propose the association of ß-glycerophosphate (ßGP) and calcium-hydroxide with chitosan (CH) to formulate a porous bioactive scaffold suitable as a cell-homing platform for dentin regeneration. METHODS: Calcium hydroxide and ßGP solutions were incorporated into chitosan to modulate scaffold architecture and composition by a phase separation technique. Architecture, chemical composition, and degradability were evaluated, and biological characterizations were performed by the seeding of dental pulp cells (DPCs) onto scaffolds, or by cultivating them in contact with leachable components (extracts), to determine cytocompatibility and odontoblastic differentiation. Cell-free scaffolds were then positioned in intimate contact with a 3D culture of DPCs in a pulp-in-a-chip platform under simulated pulp pressure. Cell mobilization and odontoblastic marker expression were evaluated. Deposition of mineralized matrix was assessed in direct contact with dentin, in the absence of osteogenic factors. RESULTS: Incorporation of calcium hydroxide and ßGP generated a stable porous chitosan scaffold containing Ca-P nanoglobule topography (CH-Ca-ßGP), which favored cell viability, alkaline phosphatase activity, and mineralized matrix deposition by cells seeded onto the scaffold structure and at a distance. The pulp-in-a-chip assay denoted its chemotactic and bioactive potential, since dentin sialoprotein-positive DPCs from 3D culture adhered to CH-Ca-ßGP more than to plain chitosan. The higher deposition of mineralized matrix onto the scaffold and surrounding dentin was also observed. SIGNIFICANCE: A CH-Ca-ßGP scaffold creates a microenvironment capable of mobilizing DPC migration toward its structure, harnessing the odontogenic potential and culminating in the expression of a highly mineralizing phenotype, key factors for a cell-homing strategy.

Effect of Time and Temperature of Air Jet on the Mechanical and Biological Behavior of a Universal Adhesive System.

OBJECTIVES: To evaluate the influence of heat application on the degree of conversion (DC) of the 3M Single Bond Universal Adhesive System, as well as its transdentinal cytotoxicity and microtensile bond strength to dentin. METHODS: Experimental groups were established according to the time and temperature of the air jet: G1: 5 seconds-25°C; G2: 10 seconds-25°C; G3: 20 seconds-25°C; G4: 5 seconds-50°C; G5: 10 seconds-50°C; G6: 20 seconds-50°C. In control group (G7), no treatment was performed. The DC was assessed using the Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) technique. For the transdentinal cytotoxicity test, dentin discs fitted in artificial pulp chambers (APC) received the application of the adhesive system and the air jets. For the microtensile bond strength, healthy molars were restored and submitted to the microtensile test after 24 hours and 6 months, respectively. RESULTS: Significant reduction in viability of Mouse Dental Papilla Cell-23 (MDPC-23), which exhibited morphological changes, was observed in all experimental groups compared to control (p<0.05). Although all tested protocols resulted in transdentinal diffusion of 2-hydroxyethyl methacrylate (HEMA), the group G6 presented the highest degree of monomeric conversion and the lowest cytotoxic effect, with higher dentin bond strength values in comparison to group G1 (p<0.05). CONCLUSIONS: Applying an air blast at 50°C for 20 seconds increases the DC and microtensile bond strength of the 3M Single Bond Universal Adhesive System to dentin, as well as reduces the transdentinal cytotoxicity of the material to pulp cells.

Influence of Manganese Oxide on the Esthetic Efficacy and Toxicity Caused by Conventional In-office Tooth Bleaching Therapy.

OBJECTIVE: This study aimed to evaluate the esthetic efficacy, cytotoxicity, and kinetics of decomposition of hydrogen peroxide (H2O2) present in a commercial bleaching gel with 35% H2O2 (BG35%) chemically activated with manganese oxide (MnO2). METHODS AND MATERIALS: After incorporating 2 mg/mL, 6 mg/mL, and 10 mg/mL of MnO2 into BG35%, the stability of pH and temperature of the products were analyzed. To assess the esthetic efficacy (ΔE and ΔWI), the BG35%s with MnO2 were applied for 45 minutes on enamel/dentin discs (DiE/D). BG35% or no treatment were used as positive (PC) and negative (NC) controls, respectively. To analyze the cell viability (CV) and oxidative stress (OXS), the same bleaching protocols were performed on DiE/D adapted to artificial pulp chambers. The extracts (culture medium + gel components that diffused through the discs) were applied to pulp cells and submitted to H2O2 quantification. BG35% with MnO2 that showed the best results was evaluated relative to kinetic decomposition of H2O2, with consequent generation of free radicals (FR) and hydroxyl radicals (OH•). The data were submitted to the one-way analysis of variance complemented by Tukey post-test (α=0.05). Data on kinetics of H2O2 decomposition were submitted to the Student's-t test (α=0.05). RESULTS: All the BG35%s with MnO2 showed stability of pH and temperature, and the gel with 10 mg/mL of this activator had an esthetic efficacy 31% higher than that of the PC (p<0.05). Reduction in OXS and trans-amelodentinal diffusion of H2O2 occurred when all the BG35%s with MnO2 were used. The addition of 6 and 10 mg/mL of MnO2 to BG35% increased the CV in comparison with PC, confirmed by the cell morphology analysis. An increase in FR and OH• formation was observed when 10 mg/mL of MnO2 was added to BG35%. CONCLUSION: Catalysis of BG35% with MnO2 minimized the trans-amelodentinal diffusion of H2O2 and toxicity of the product to pulp cells. BG35% containing 10 mg/mL of MnO2 potentiated the decomposition of H2O2, enhancing the generation of FR and OH•, as well as the efficacy of the in-office tooth therapy.

Does silver diammine fluoride concentration influence on bonding to carious dentinal lesions in primary teeth?

PURPOSE: This in vitro study evaluated the microshear bond strength (µSBS) of a universal adhesive after silver diammine fluoride (SDF) application at different concentrations on carious dentinal lesions in primary teeth. METHODS: Flat dentin carious-induced surfaces from 40 primary molars were randomly assigned to four experimental groups: without SDF treatment (control), 12% SDF, 30% SDF, or 38% SDF application. After 14 days of storage in artificial saliva at 37 °C, a universal adhesive system (Scotchbond Universal, 3 M ESPE) was applied in the etch-and-rinse mode and resin composite cylinders were built (0.72 mm2). After 24 h of water storage, the µSBS test was performed and the failure mode was determined. Data were analysed using one-way ANOVA and Tukey's post hoc tests (α = 0.05). RESULTS: The application of 38% SDF resulted in higher µSBS mean than control and use of 12% SDF (p = 0.006). No significant differences were observed among control, 12% SDF, and 30% SDF. All specimens tested showed adhesive/mixed failure. CONCLUSION: The use of SDF does not jeopardise the bonding of a universal adhesive applied in the etch-and-rinse mode to carious dentinal lesions in primary teeth, irrespective of the product concentration.

Terahertz radiation from bacteriorhodopsin reveals correlated primary electron and proton transfer processes.

The kinetics of electrogenic events associated with the different steps of the light-induced proton pump of bacteriorhodopsin is well studied in a wide range of time scales by direct electric methods. However, the investigation of the fundamental primary charge translocation phenomena taking place in the functional energy conversion process of this protein, and in other biomolecular assemblies using light energy, has remained experimentally unfeasible because of the lack of proper detection technique operating in the 0.1- to 20-THz region. Here, we show that extending the concept of the familiar Hertzian dipole emission into the extreme spatial and temporal range of intramolecular polarization processes provides an alternative way to study ultrafast electrogenic events on naturally ordered biological systems. Applying a relatively simple experimental arrangement based on this idea, we were able to observe light-induced coherent terahertz radiation from bacteriorhodopsin with femtosecond time resolution. The detected terahertz signal was analyzed by numerical simulation in the framework of different models for the elementary polarization processes. It was found that the principal component of the terahertz emission can be well described by excited-state intramolecular electron transfer within the retinal chromophore. An additional slower process is attributed to the earliest phase of the proton pump, probably occurring by the redistribution of a H bond near the retinal. The correlated electron and proton translocation supports the concept, assigning a functional role to the light-induced sudden polarization in retinal proteins.

Transenamel and transdentinal cytotoxicity of carbamide peroxide bleaching gels on odontoblast-like MDPC-23 cells.

AIM: To evaluate the transenamel and transdentinal cytotoxicity of bleaching gels based on carbamide peroxide (CP) on odontoblast-like cells after different contact times of the products with enamel. METHODOLOGY: Enamel/dentine discs were obtained from bovine incisors and placed in artificial pulp chambers. Bleaching gels containing 10% or 16% CP were applied for 8 h day(-1) on the enamel side of the discs during periods of 1, 7 or 14 days. Deionized water and artificial saliva served as controls. The extracts (culture medium plus bleaching gel products that diffused through the discs) were collected and applied on previously cultured MDPC-23 cells for 1 h. Cell metabolism was evaluated by the MTT assay, and the data were analysed statistically by one-way anova and Tukey's test (α=0.05). Cell morphology was analysed by SEM. RESULTS: There was no significant difference (P>0.05) between the controls and the groups bleached with 10% CP gel. In the groups bleached with 16% CP gel, however, cell metabolism decreased significantly (P<0.05) by 40.32%, 30.16% and 26.61% at 1, 7 and 14 days, respectively. There was no significant difference (P>0.05) between 1, 7 or 14 applications of the gels for either of the CP concentrations. CONCLUSION: Regardless of the number of applications on an enamel surface, the 10% CP bleaching gel did not cause transenamel and transdentinal cytotoxicity to the MDPC-23 cell cultures. However, diffusion of products from the 16% CP gel through enamel and dentine and cytopathic effects to the pulp cells occurred even after a single application of this product on enamel.

Mechanical Stability and Proteolytic Activity of Resin-dentin Bonds Using the Cross-linked Dry Bonding Technique.

OBJECTIVE: To evaluate the mechanical stability and the proteolytic activity of bonds created by a two-step, etch-and-rinse adhesive applied to cross-linked and air-dried etched dentin. METHODS: Flat dentin surfaces were produced in 64 extracted sound human molars. The dentin was etched with 35% phosphoric acid for 15 seconds, and then the teeth were divided into groups according to the cross-linking solution applied on the etched dentin. Group 1: 5% grape seed extract (GSE), Group 2: 5% glutaraldehyde, Group 3: Gluma Desensitizer, or Group 4: deionized water (control). Solutions were applied for 60 seconds, followed by rinse and blot drying. Then, the teeth were separated into two subgroups where the etched dentin was kept moist or air-dried. The adhesive was applied followed by a composite resin buildup. After 24 hours, the teeth were cut into beams (0.81 mm²) that were tested for microtensile strength immediately or after 12 months of aging in a 37°C saliva-like buffer. Additional teeth (n=32) were bonded as described and cut into 0.5-mm-thick slabs. The slabs were prepared for nanoleakage (scanning electron microscopy) and in situ zymography (EnzChek Protease Assay Kit). Bond strength data were submitted to ANOVA and Tukey tests (α =0.05). RESULTS: Significant reduction in immediate bond strength (ca 65%) and increase in proteolytic activity was seen when the etched dentin was air dried without previous cross-linking biomodification. Conversely, bond strengths did not differ from those produced on wet dentin when collagen was cross-linked before air drying, irrespective of the solution applied. For both moist and air-dried etched dentin, collagen cross-linking resulted in mechanically stable bonds and reduced proteolytic activity after 12 months of storage. CONCLUSION: Bonds produced by the application of a two-step, etch-and-rinse adhesive to cross-linked, air-dried, etched dentin were mechanically stable and revealed reduced proteolytic activity after 1 year of aging.

Chitosan-Calcium-Simvastatin Scaffold as an Inductive Cell-Free Platform.

The development of biomaterials based on the combination of biopolymers with bioactive compounds to develop delivery systems capable of modulating dentin regeneration mediated by resident cells is the goal of current biology-based strategies for regenerative dentistry. In this article, the bioactive potential of a simvastatin (SV)-releasing chitosan-calcium-hydroxide (CH-Ca) scaffold was assessed. After the incorporation of SV into CH-Ca, characterization of the scaffold was performed. Dental pulp cells (DPCs) were seeded onto scaffolds for the assessment of cytocompatibility, and odontoblastic differentiation was evaluated in a microenvironment surrounded by dentin. Thereafter, the cell-free scaffold was adapted to dentin discs positioned in artificial pulp chambers in direct contact with a 3-dimensional (3D) culture of DPCs, and the system was sealed to simulate internal pressure at 20 cm/H2O. In vivo experiments with cell-free scaffolds were performed in rats' calvaria defects. Fourier-transform infrared spectroscopy spectra proved incorporation of Ca and SV into the scaffold structure. Ca and SV were released upon immersion in a neutral environment. Viable DPCs were able to spread and proliferate on the scaffold over 14 d. Odontoblastic differentiation occurred in the DPC/scaffold constructs in contact with dentin, in which SV supplementation promoted odontoblastic marker overexpression and enhanced mineralized matrix deposition. The chemoattractant potential of the CH-Ca scaffold was improved by SV, with numerous viable and dentin sialoprotein-positive cells from the 3D culture being observed on its surface. Cells at 3D culture featured increased gene expression of odontoblastic markers in contact with the SV-enriched CH-Ca scaffold. CH-Ca-SV led to intense mineralization in vivo, presenting mineralization foci inside its structure. In conclusion, the CH-Ca-SV scaffold induces differentiation of DPCs into a highly mineralizing phenotype in the presence of dentin, creating a microenvironment capable of attracting pulp cells to its surface and inducing the overexpression of odontoblastic markers in a cell-homing strategy.