Possibility of Human Gender Recognition Using Raman Spectra of Teeth.

Gender determination of the human remains can be very challenging, especially in the case of incomplete ones. Herein, we report a proof-of-concept experiment where the possibility of gender recognition using Raman spectroscopy of teeth is investigated. Raman spectra were recorded from male and female molars and premolars on two distinct sites, tooth apex and anatomical neck. Recorded spectra were sorted into suitable datasets and initially analyzed with principal component analysis, which showed a distinction between spectra of male and female teeth. Then, reduced datasets with scores of the first 20 principal components were formed and two classification algorithms, support vector machine and artificial neural networks, were applied to form classification models for gender recognition. The obtained results showed that gender recognition with Raman spectra of teeth is possible but strongly depends both on the tooth type and spectrum recording site. The difference in classification accuracy between different tooth types and recording sites are discussed in terms of the molecular structure difference caused by the influence of masticatory loading or gender-dependent life events.

Comparability of Raman Spectroscopic Configurations: A Large Scale Cross-Laboratory Study.

The variable configuration of Raman spectroscopic platforms is one of the major obstacles in establishing Raman spectroscopy as a valuable physicochemical method within real-world scenarios such as clinical diagnostics. For such real world applications like diagnostic classification, the models should ideally be usable to predict data from different setups. Whether it is done by training a rugged model with data from many setups or by a primary-replica strategy where models are developed on a 'primary' setup and the test data are generated on 'replicate' setups, this is only possible if the Raman spectra from different setups are consistent, reproducible, and comparable. However, Raman spectra can be highly sensitive to the measurement conditions, and they change from setup to setup even if the same samples are measured. Although increasingly recognized as an issue, the dependence of the Raman spectra on the instrumental configuration is far from being fully understood and great effort is needed to address the resulting spectral variations and to correct for them. To make the severity of the situation clear, we present a round robin experiment investigating the comparability of 35 Raman spectroscopic devices with different configurations in 15 institutes within seven European countries from the COST (European Cooperation in Science and Technology) action Raman4clinics. The experiment was developed in a fashion that allows various instrumental configurations ranging from highly confocal setups to fibre-optic based systems with different excitation wavelengths. We illustrate the spectral variations caused by the instrumental configurations from the perspectives of peak shifts, intensity variations, peak widths, and noise levels. We conclude this contribution with recommendations that may help to improve the inter-laboratory studies.

Light transmittance and polymerization kinetics of amorphous calcium phosphate composites.

OBJECTIVES: This study investigated light transmittance and polymerization kinetics of experimental remineralizing composite materials based on amorphous calcium phosphate (ACP), reinforced with inert fillers. MATERIALS AND METHODS: Light-curable composites were composed of Bis-EMA-TEGDMA-HEMA resin and ACP, barium glass, and silica fillers. Additionally, a commercial composite Tetric EvoCeram was used as a reference. Light transmittance was recorded in real-time during curing, and transmittance curves were used to assess polymerization kinetics. To obtain additional information on polymerization kinetics, temperature rise was monitored in real-time during curing and degree of conversion was measured immediately and 24 h post-cure. RESULTS: Light transmittance values of 2-mm thick samples of uncured ACP composites (2.3-2.9 %) were significantly lower than those of the commercial composite (3.8 %). The ACP composites presented a considerable transmittance rise during curing, resulting in post-cure transmittance values similar to or higher than those of the commercial composite (5.5-7.9 vs. 5.4 %). The initial part of light transmittance curves of experimental composites showed a linear rise that lasted for 7-20 s. Linear fitting was performed to obtain a function whose slope was assessed as a measure of polymerization rate. Comparison of transmittance and temperature curves showed that the linear transmittance rise lasted throughout the most part of the pre-vitrification period. CONCLUSIONS: The linear rise of light transmittance during curing has not been reported in previous studies and may indicate a unique kinetic behavior, characterized by a long period of nearly constant polymerization rate. CLINICAL RELEVANCE: The observed kinetic behavior may result in slower development of polymerization shrinkage stress but also inferior mechanical properties.

FTIR spectroscopy reveals lipid droplets in drug resistant laryngeal carcinoma cells through detection of increased ester vibrational bands intensity.

The major obstacle to successful chemotherapy of cancer patients is drug resistance. Previously we explored the molecular mechanisms of curcumin cross-resistance in carboplatin resistant human laryngeal carcinoma 7T cells. Following curcumin treatment we found a reduction in curcumin accumulation, and reduced induction of reactive oxygen species (ROS) and their downstream effects, compared to parental HEp-2 cells. In order to shed more light on mechanisms involved in drug resistance of 7T cells, in the present study we applied Fourier transform infrared (FTIR) spectroscopy, a technique that provides information about the nature and quantities of all molecules present in the cell. By comparing the spectra from parental HEp-2 cells and their 7T subline, we found an increase in the intensity of ester vibrational bands in 7T cells. This implied an increase in the amount of cholesteryl esters in resistant cells, which we confirmed by an enzymatic assay. Since cholesteryl esters are localized in lipid droplets, we confirmed their higher quantity and serum dependency in 7T cells compared to HEp-2 cells. Moreover, treatment with oleic acid induced more lipid droplets in 7T when compared to HEp-2 cells, as shown by flow cytometry. We can conclude that along with previously determined molecular mechanisms of curcumin resistance in 7T cells, these cells exhibit an increased content of cholesteryl esters and lipid droplets, suggesting an alteration in cellular lipid metabolism as a possible additional mechanism of drug resistance. Furthermore, our results suggest the use of FTIR spectroscopy as a promising technique in drug resistance research.

Influence of One-Dimensional Photonic Crystal on Raman Signal Enhancement: A Detailed Experimental Study.

The enhancement of Raman signals using photonic crystal structures has been the subject of numerous experimental and theoretical studies, leading to a variety of issues and inconsistencies. This paper presents a comprehensive experimental investigation into the impact of alignment between the laser excitation wavelength and the specific position of the photonic band gap on signal enhancement in Raman spectroscopy. By employing one-dimensional (1D) porous silicon photonic crystals, a systematic analysis across a large number of spectra was conducted. The study focused on examining the signal enhancement of both the Raman ∼520 cm-1 silicon band, representing the constituent material of photonic crystal, and the most prominent Raman bands of crystal violet, used as a probe molecule. The probe molecules were both infiltrated into and adsorbed on top of the photonic crystal structure. The obtained experimental results for the contribution of 1D photonic crystals to Raman signal enhancement are much smaller compared to most predictions. The Raman signal of silicon and the signal from the probe molecule are enhanced ≤2.5 times when the laser excitation aligns with the edge of the photonic band gap, strictly defined as the position at the very bottom of the reflectance peak. The results have been discussed within the context of theoretical explanations.

Age Estimation and Sex Determination Using Raman Spectra of Human Dentine.

Objectives: This study aimed to determine the sex and estimate the age of individuals by analysing Raman spectra obtained from extracted teeth dentine. Material and methods: A total of 25 male and 26 female extracted teeth were collected, disinfected, and fixed in auto acrylate before being cut near the cervical margin. The inclusion criteria were non endodontically treated teeth with the cervical third of the tooth crown preserved. The exposed dentine was recorded using a Raman spectrometer, with 100 scans per dentine site in the spectral range of 3500 to 200 cm-1 and a resolution of 4 cm-1. Background-corrected, normalized, and decomposed spectra were subjected to principal component analysis using MATLAB 2010 (The MathWorks, Natick, MA, USA). To assess age differences, prominent vibrational bands were observed and statistically analyzed, while sex differentiation employed the T-test on acquired data, exploring vibrational bands with significant intensity differences. Results: The obtained results revealed a segmentation of spectra in the male sample group based on the age of teeth at extraction, with an accuracy of age estimation at 7.0048 years. A similar segmentation was observed in the female sample group, with an accuracy of 9.3863 years. Conclusion: The recorded spectra were organized into datasets and analyzed with principal component analysis, demonstrating the feasibility of sex determination and age estimation using Raman spectra of teeth. Differences in classification accuracy between sexes may be attributed to hormonally-mediated differences in the biochemical composition of dentine between males and females. The findings of this study can provide valuable insights into methods for forensic and anthropological applications.

Fourier Transform Infrared Spectroscopy Reveals Molecular Changes in Blood Vessels of Rats Treated with Pentadecapeptide BPC 157.

Recently, it was found that when confronted with major vessel occlusion and vascular failure, stable gastric pentadecapeptide BPC 157 therapy might rapidly functionally improve minor vessels to take over the function of disabled major vessels, reorganize blood flow, and compensate failed vessel function. We focused on the BPC 157 therapy effect obtained by giving 10 ng/kg ip to rats 5 min before sacrifice on the rat thoracic aorta, which we assessed with Fourier transform infrared spectroscopy (FTIR) 90 min thereafter. We applied a principal component analysis (PCA). The PCA model showed, with a clear distinction being mostly due to the PC1 score, differences between the spectra of BPC 157- and saline-treated rats. The comparison of the averaged spectra of these two groups with their differential spectrum and PC loadings allowed us to identify the parts of the FTIR spectra that contributed the most to the spectral separation of the two observed groups. The PC1 loadings and the differential spectrum showed that the main bands affecting the separation were the amid I band around 1650 cm-1, the amid II band around 1540 cm-1, and the vibrational band around 1744 cm-1. Fitting the spectral range between 1450 and 1800 cm-1 showed changes in protein conformation and confirmed the appearance of the vibrational band at 1744 cm-1. Controls had a substantially more intense vibrational band at 1744 cm-1. These spectral results showed the cells from saline-treated (control) rats to be in the early stage of cell death, while the samples from BPC 157-rats were protected. Thus, BPC 157 therapy changed the lipid contents and protein secondary structure conformation, with a rapid effect on vessels, within a short time upon application.

Rapid 3 s Curing: What Happens in Deep Layers of New Bulk-Fill Composites?

This study assessed the influence of rapid 3 s light curing on the new generation of bulk-fill resin composites under the simulated aging challenge and depths up to 4 mm. Four bulk-fill materials were tested: two materials designed for rapid curing (Tetric PowerFill-PFILL; Tetric PowerFlow-PFLW) and two regular materials (Filtek One Bulk Fill Restorative-FIL; SDR Plus Bulk Fill Flowable-SDR). Three-point bending (n = 10) was used to measure flexural strength (FS) and flexural modulus (FM). In the 3 s group, two 2 mm thick specimens were stacked to obtain 4 mm thickness, while 2 mm-thick specimens were used for ISO group. Specimens were aged for 1, 30, or 30 + 3 days in ethanol. The degree of conversion (DC) up to 4 mm was measured by Raman spectroscopy. There was no difference between curing protocols in FS after 1 day for all materials except PFLW. FM was higher for all materials for ISO curing protocol. Mechanical properties deteriorated by increasing depth (2-4 mm) and aging. ISO curing induced higher DC for PFLW and FIL, while 3 s curing was sufficient for PFILL and SDR. The 3 s curing negatively affected FM of all tested materials, whereas its influence on FS and DC was highly material-specific.

Principal Component Regression for Forensic Age Determination Using the Raman Spectra of Teeth.

Raman spectra of mineralized tooth tissues were used to build a principal component regression (PCR) age determination model for forensic application. A sample of 71 teeth was obtained from donors aging from 11 to 76 years. No particular selection criteria were applied; teeth affected with various pathological processes were deliberately included to simulate a realistic forensic scenario. In order to comply with the nondestructive specimen handling, Raman spectra were collected from tooth surfaces without any previous preparation. Different tooth tissues were evaluated by collecting the spectra from three distinct sites: tooth crown, tooth neck, and root apex. Whole recorded spectra (3500-200 cm-1) were used for principal component analysis and building of the age determination model using PCR. The predictive capabilities of the obtained age determination models varied according to the spectra collection site. Optimal age determination was attained by using Raman spectra collected from cementum at root apex (R2 values of 0.84 and 0.71 for male and female donors, respectively). For optimal performance of that model, male and female donors had to be analyzed separately, as merging both genders into a single model considerably diminished its predictive capability (R2 = 0.29).

Differentiating Between Malignant Mesothelioma and Other Pleural Lesions Using Fourier Transform Infrared Spectroscopy.

Histopathology, despite being the gold standard as a diagnostic tool, does not always provide a correct diagnosis for different pleural lesions. Although great progress was made in this field, the problem to differentiate between reactive and malignant pleural lesions still stimulates the search for additional diagnostic tools. Our research using vibrational spectroscopy and principal component analysis (PCA) statistical modeling represents a potentially useful tool to approach the problem. The objective method this paper explores is based on the correlation between different types of pleural lesions and their vibrational spectra. Obtained tissue spectra recorded by infrared spectroscopy allowed us to categorize spectra in different groups using a created PCA statistical model. The PCA model was built using tissues of known pathology as the model group. The validation samples were then used to confirm the functionality of our PCA model. Student's t-test was also used for comparing samples in paired groups. The PCA model was able to clearly differentiate the spectra of mesothelioma, metastasis and reactive changes (inflammation), and place them in discrete groups. Thus, we showed that Fourier transform infrared spectroscopy combined with PCA can differentiate pleural lesions with high sensitivity and specificity. This new approach could contribute in objectively differentiating specific pleural lesions, thus helping pathologists to better diagnose difficult pleural samples but also could shed additional light into the biology of malignant pleural mesothelioma.

Embryo-derived teratoma in vitro biological system reveals antitumor and embryotoxic activity of valproate.

Antiepileptic/teratogen valproate (VPA) is a histone deacetylase inhibitor/epigenetic drug proposed for the antitumor therapy where it is generally crucial to target poorly or undifferentiated cells to prevent a recurrence. Transplanted rodent gastrulating embryos-proper (primitive streak and three germ layers) are the source of teratoma/teratocarcinoma tumors. Human primitive-streak remnants develop sacrococcygeal teratomas that may recur even when benign (well differentiated). To screen for unknown VPA impact on teratoma-type tumors, we used original 2-week embryo-derived teratoma in vitro biological system completed by a spent media metabolome analysis. Gastrulating 9.5-day-old rat embryos-proper were cultivated in Eagle's minimal essential medium (MEM) with 50% rat serum (controls) or with the addition of 2 mmVPA. Spent media metabolomes were analyzed by FTIR. Compared to controls, VPA acetylated histones; significantly diminished overall teratoma growth, impaired survival, increased the apoptotic index, and decreased proliferation index and incidence of differentiated tissues (e.g., neural tissue). Control teratomas continued to grow and differentiate for 14 days in isotransplants in vivo, but in vitro VPA-treated teratomas resorbed. Principal component analysis of FTIR results showed that spent media metabolomes formed well-separated clusters reflecting the treatment and day of cultivation. In metabolomes of VPA-treated teratomas, we found elevation of previously described histone acetylation biomarkers [amide I α-helix and A(CH3 )/A(CH2 )]) with apoptotic biomarkers within the amide I region for ß-sheets, and unordered and CH2 vibrations of lipids. VPA may be proposed for therapy of the undifferentiated component of teratoma tumors and this biological system completed by metabolome analysis, for a faster dual screening of antitumor/embryotoxic agents.

The effect of excitation laser power in Raman spectroscopic measurements of the degree of conversion of resin composites.

OBJECTIVES: To evaluate the effect of excitation laser power in Raman spectrometry by comparing the spectra and the degree of conversion (DC) values obtained using excitation powers between 300 and 1000mW. METHODS: Five commercial and three experimental resin composites were light cured at 1200mW/cm2 for 10-20s from a commercial blue-violet LED dental curing unit. Raman spectra were collected from composite specimens within 9min after light-curing. The excitation laser (1064nm) was focused on the spot of 0.4mm in diameter. The following powers were used for specimen excitation (mW): 300, 400, 600, 800, and 1000. From Raman spectra, the DC values were calculated and compared among different laser powers. Also, vector-normalized Raman spectra collected using the lowest excitation power (300mW) were compared to those collected using the maximum excitation power (1000mW). RESULTS: Varying the excitation laser power between 300 and 1000mW resulted in statistically significant differences in both the DC values and the intensity of particular spectral features. The effect of varying laser power on Raman spectra and obtained DC values was material-dependent. The DC values measured within an individual material using different laser powers varied between 3.2 and 7.2% (absolute DC difference). The spectral bands affected by variations in laser power were assigned to symmetric and asymmetric stretching of -CH2 (2900-3100cm-1), symmetric stretching of aliphatic CC (1640cm-1) and scissoring of C-H (1458cm-1). SIGNIFICANCE: The DC can be artificially elevated through increasing excitation laser power. This effect should be considered in Raman spectroscopic evaluations of DC in specimens during ongoing post-cure polymerization.

Long-term water sorption and solubility of experimental bioactive composites based on amorphous calcium phosphate and bioactive glass.

The aim of this study was to evaluate water sorption and solubility of two series of experimental composites containing amorphous calcium phosphate (ACP) or bioactive glass (BG). Water sorption and solubility were measured for up to 287 days. The surface precipitation of calcium phosphates was evaluated by scanning electron microscopy. The ACP-series showed higher water sorption (223-568 µg/mm3) than the BG-series (40-232 µg/mm3). In contrast, the ACP-series had generally lower solubility (37-106 µg/mm3) than the BG-series (1-506 µg/mm3). The constant specimen mass for the ACP-series was attained after 14 days of water immersion, while the mass decrease due to long-term solubility in the BG-series lasted beyond 287 days. Calcium phosphates precipitated in composites with the BG filler loading of 10 wt% or more, as well as in all of the ACP-containing composites. The experimental composite series showed water sorption and solubility considerably higher than commercial materials.

Near-Infrared Surface-Enhanced Raman Scattering on Silver-Coated Porous Silicon Photonic Crystals.

Surface-enhanced Raman scattering (SERS) with near-infrared (NIR) excitation offers a safe way for the detection and study of fragile biomolecules. In this work, we present the possibility of using silver-coated porous silicon photonic crystals as SERS substrates for near-infrared (1064 nm) excitation. Due to the deep penetration of NIR light inside silicon, the fabrication of photonic crystals was necessary to quench the band gap photoluminescence of silicon crystal, which acts as mechanical support for the porous layer. Optimal parameters of the immersion plating process that gave maximum enhancement were found and the activity of SERS substrates was tested using rhodamine 6G and crystal violet dye molecules, yielding significant SERS enhancement for off-resonant conditions. To our knowledge, this is the first time that the 1064 nm NIR laser excitation is used for obtaining the SERS effect on porous silicon as a substrate.

Curing potential of experimental resin composites with systematically varying amount of bioactive glass: Degree of conversion, light transmittance and depth of cure.

OBJECTIVES: The aim of this work was to investigate the curing potential of an experimental resin composite series with the systematically varying amount of bioactive glass 45S5 by evaluating the degree of conversion, light transmittance and depth of cure. METHODS: Resin composites based on a Bis-GMA/TEGDMA resin with a total filler load of 70 wt% and a variable amount of bioactive glass (0-40 wt%) were prepared. The photoinitiator system was camphorquinone and ethyl-4-(dimethylamino) benzoate. The degree of conversion and light transmittance were measured by Raman spectroscopy and UV-vis spectroscopy, respectively. The depth of cure was evaluated according to the classical ISO 4049 test. RESULTS: The initial introduction of bioactive glass into the experimental series diminished the light transmittance while the further increase in the bioactive glass amount up to 40 wt% caused minor variations with no clear trend. The curing potential of the experimental composites was similar to or better than that of commercial resin composites. However, unsilanized bioactive glass fillers demonstrated the tendency to diminish both the maximum attainable conversion and the curing efficiency at depth. CONCLUSIONS: Experimental composite materials containing bioactive glass showed a clinically acceptable degree of conversion and depth of cure. The degree of conversion and depth of cure were diminished by bioactive glass fillers in a dose-dependent manner, although light transmittance was similar among all of the experimental composites containing 5-40 wt% of bioactive glass. CLINICAL SIGNIFICANCE: Reduced curing potential caused by the bioactive glass has possible consequences on mechanical properties and biocompatibility.

Analysis of pulmonary surfactant by Fourier transform infrared spectroscopy after exposure to sevoflurane and isoflurane.

Pulmonary surfactant, consisting primarily of phospholipids and four surfactant-specific proteins, is among the first structures that is exposed to inhalation anesthetics. Consequently, changes of pulmonary surfactant due to this exposure could cause respiratory complications after long anesthetic procedures. Fourier transform infrared (FTIR) spectroscopy was used to explore the effects of two inhalation anesthetics, sevoflurane and isoflurane, on a commercially available pulmonary surfactant. The research was primarily focused on the effect of anesthetics on the lipid component of the surfactant. Four different concentrations of anesthetics were added, and the doses were higher from the low clinical doses typically used. Recorded spectra were analyzed using principal component analysis, and the Student's t-test was performed to confirm the results. The exposure to both anesthetics induced similar changes, consistent with the increase of the anesthetic concentration. The most pronounced effect was on the hydrophilic head group of phospholipids, which is in agreement with the disruption of the hydrogen bond, caused by the anesthetics. A change in the band intensities of CH2 stretching vibrations, indicative of a disordering effect of anesthetics on the hydrophobic tails of phospholipids, was also observed. Changes induced by isoflurane appear to be more pronounced than those induced by sevoflurane. Furthermore, our results suggest that FTIR spectroscopy is a promising tool in studying anesthetic effects on pulmonary surfactant.

Conversion and temperature rise of remineralizing composites reinforced with inert fillers.

OBJECTIVES: Remineralizing experimental composites based on amorphous calcium phosphate (ACP) were investigated. The impact of curing time (20 and 40s), curing depth (1, 2, 3 and 4mm) and addition of inert fillers (barium glass and silica) on the conversion and temperature rise during curing were examined. METHODS: Five ACP-composites and two control composites were prepared based on the light-curable EBPADMA-TEGDMA-HEMA resin. For temperature measurements, a commercial composite was used as an additional control. Conversion was assessed using FT-Raman spectroscopy by comparing the relative change of the band at 1640 cm(-1) before and after polymerization. The temperature rise during curing was recorded in real-time using a T-type thermocouple. RESULTS: At 1mm depth, the ACP-composites attained significantly higher conversion (77.8-87.3%) than the control composites based on the same resin (60.5-66.3%). The addition of inert fillers resulted in approximately 5% lower conversion at clinically relevant depths (up to 2mm) for the curing time of 40s. Conversion decline through depths depended on the added inert fillers. Conversion values higher than 80% of the maximum conversion were observed for all of the ACP-composites at depths up to 3mm, when cured for 40s. Significantly higher total temperature rise for the ACP-composites (11.5-13.1 °C) was measured compared to the control composites (8.6-10.8 °C) and the commercial control (8.7 °C). CONCLUSIONS: The admixture of inert fillers represents a promising strategy for further development of ACP-composites, as it reduced the temperature rise while negligibly impairing the conversion. CLINICAL SIGNIFICANCE: High conversions of ACP-composites are favorable in terms of mechanical properties and biocompatibility. However, high conversions were accompanied with high temperature rise, which might present a pulpal hazard.

Impedance changes during setting of amorphous calcium phosphate composites.

OBJECTIVES: To investigate the electrical properties of experimental light-curable composite materials based on amorphous calcium phosphate (ACP) with the admixture of silanized barium glass and silica fillers. METHODS: Short-term setting was investigated by impedance measurements at a frequency of 1kHz, while for the long-term setting the impedance spectra were measured consecutively over a frequency range of 0.05Hz to 1MHz for 24h. The analysis of electrical resistivity changes during curing allowed the extraction of relevant kinetic parameters. The impedance results were correlated to the degree of conversion assessed by Raman spectroscopy, water content determined by gravimetry, light transmittance measured by CCD spectrometer and microstructural features observed by scanning electron microscopy. RESULTS: ACP-based composites have shown higher immediate degree of conversion and less post-cure polymerization than the control composites, but lower polymerization rate. The polymerization rate assessed by impedance measurements correlated well with the light transmittance. The differences in the electrical conductivity values observed among the materials were correlated to the amount of water introduced into composites by the ACP filler. High correlation was found between the degree of conversion and electrical resistivity. Equivalent circuit modeling revealed two electrical contributions for the ACP-based composites and a single contribution for the control composites. SIGNIFICANCE: The impedance spectroscopy has proven a valuable method for gaining insight into various features of ACP-based composites. Better understanding of the properties of ACP-based composites should further the development of these promising bioactive materials.

Long Term Degree of Conversion of two Bulk-Fill Composites.

OBJECTIVES: To investigate the long-term development of the post-cure degree of conversion (DC) for two flowable bulk-fill composites. MATERIALS AND METHODS: Tetric EvoFlow Bulk Fill (TEFBF) and SDR were chosen due to their distinct compositional modifications that enable the decrease of translucency during polymerization and lower polymerization rate, respectively. DC was assessed using FT-Raman spectroscopy at the post-cure times of 0 h, 24 h, 7 d and 30 d. The post-cure behavior was analyzed by a mixed model ANOVA and partial eta-squared statistics. RESULTS: DC ranged from 61.3-81.1% for TEFBF and 58.9-81.6% for SDR. The initial (0 h) DC was significantly lower at a depth of 4 mm than at a depth of 1 mm (4.9% for SDR and 11.1% for TEFBF). Both materials presented a significant post-cure DC increase, up to 16.4% for TEFBF and 20.6% for SDR. The post-cure DC development was depth-dependent for TEFBF, but not for SDR. The post-cure DC increase was observed during 24 h for TEFBF and 7 d for SDR. CONCLUSIONS: Some of the bulk-fill composites may need longer times than the commonly accepted 24 h to reach the final conversion. This may be attributed to their compositional modifications that are mostly undisclosed by manufacturers. Our findings imply that investigations commonly performed 24 h post-cure may underestimate some of the bulk-fill composite properties, if these are affected by the slowly-developing DC. Reactive species may also be available for leaching out of the restoration during an extended time period, with possible implications on biocompatibility.

Porous Silicon Covered with Silver Nanoparticles as Surface-Enhanced Raman Scattering (SERS) Substrate for Ultra-Low Concentration Detection.

Microporous and macro-mesoporous silicon templates for surface-enhanced Raman scattering (SERS) substrates were produced by anodization of low doped p-type silicon wafers. By immersion plating in AgNO3, the templates were covered with silver metallic film consisting of different silver nanostructures. Scanning electron microscopy (SEM) micrographs of these SERS substrates showed diverse morphology with significant difference in an average size and size distribution of silver nanoparticles. Ultraviolet-visible-near-infrared (UV-Vis-NIR) reflection spectroscopy showed plasmonic absorption at 398 and 469 nm, which is in accordance with the SEM findings. The activity of the SERS substrates was tested using rhodamine 6G (R6G) dye molecules and 514.5 nm laser excitation. Contrary to the microporous silicon template, the SERS substrate prepared from macro-mesoporous silicon template showed significantly broader size distribution of irregular silver nanoparticles as well as localized surface plasmon resonance closer to excitation laser wavelength. Such silver morphology has high SERS sensitivity that enables ultralow concentration detection of R6G dye molecules up to 10(-15) M. To our knowledge, this is the lowest concentration detected of R6G dye molecules on porous silicon-based SERS substrates, which might even indicate possible single molecule detection.