Design of colorimetric nanostructured sensor phases for simple and fast quantification of low concentrations of acid vapors.

Two colorimetric nanostructured sensor phases (Color-NSPs) for the determination of low concentrations of acid vapors in the atmosphere of paper storage rooms have been designed and characterized. The acid vapor determination is based on the color change that occurs in polyaniline (PANI) in the presence of acids when it goes from its emeraldine base form (blue) to its emeraldine salt form (green). To synthesize the Color-NSPs, two methods have been used, a one-step method performed by grafting polyaniline onto a cellulose membrane (Cellu-PANI) and a two-step method in which in the first step, polyaniline is grafted onto the surface of polymeric nanoparticles (NPs-PANI), and in a second step, NPs-PANI are immobilized into the pores of a nylon membrane (Nylon-NPs PANI). The response of the sensors versus acid vapor was measured by color coordinates with a photographic camera. A linear response range from 1 ppmv to 7 ppmv was found for both sensors, and the detection limits were 0.95 ppmv (1.2 % RSD) and 0.40 ppmv (0.8 % RSD) for Cellu-PANI and Nylon-NPs PANI, respectively. In addition, both sensors showed complete reversibility and a short exposition time (5 min). The potential applicability of the Color-NSPs in the control of the exposure of paper heritage collections to outdoor- and indoor-generated gaseous pollutants was demonstrated by determining acid vapors in museums. The method was validated with an external reference method; the paired test was applied, and p-values greater than 5% were obtained, indicating an excellent correlation and showing that the Color-NSPs reported are simple, fast, and an economical alternative to control and protect cultural heritage materials in indoor environments.

Doxycycline-functionalized polymeric nanoparticles inhibit Enterococcus faecalis biofilm formation on dentine.

AIM: To evaluate in a laboratory setting the antimicrobial properties and the potential to inhibit biofilm formation of novel remineralizing polymeric nanoparticles (NPs) when applied to dentine surfaces and to ascertain the effect of the functionalization of these NPs with zinc, calcium or doxycycline. METHODOLOGY: The antimicrobial activity and inhibition of biofilm formation of polymeric NPs were analysed on human dentine blocks that were infected with Enterococcus faecalis before or after application of NPs. LIVE/DEAD ® testing under Confocal Laser Scanning Microscopy and bacterial culturing were employed to analyse biofilm biovolume and bacterial viability. Field Emission Scanning Electron Microscopy was also employed to assess biofilm morphology. One-way anova with Welch's correction and post hoc comparison by the Games-Howell test were performed for comparisons between groups. RESULTS: The un-functionalized NPs displayed the greatest antimicrobial activity against E. faecalis biofilms as they provided the lowest biovolume (3865.7 ± 2926.97 µm3 ; P < 0.001) and the highest dead/injured cells percentage (79.93 ± 18.40%; P < 0.001), followed by Dox-NPs (biovolume: 19,041.55 ± 17,638.23 µm3 , dead/injured cells: 45.53 ± 26.50%; P < 0.001). Doxycycline-loaded NPs had the largest values of inhibition of biofilm formation with the lowest biofilm biovolume (8517.65 ± 7055.81 µm3 ; P < 0.001) and a high dead/injured bacterial percentage (68.68 ± 12.50%; P < 0.001). Un-functionalized NPs did not reduce biomass growth (P > 0.05), but attained the largest percentage of compromised cells (93 ± 8.23%; P < 0.001), being able to disrupt biofilm formation. It also produced occlusion of dentinal tubules, potentially interfering with bacterial tubule penetration. CONCLUSIONS: A new generation of bioactive nano-fillers (doxycycline-functionalized polymeric NPs) had antibacterial activity and occluded dentinal tubules. Incorporating these NPs into endodontic sealers may have the potential to enhance the outcome of root canal treatment.

Ion-modified nanoparticles induce different apatite formation in cervical dentine.

AIM: To investigate if crystallinity and ultrastructure are modified when cervical dentine is treated with four different nanogels-based solutions for remineralizing purposes. METHODOLOGY: Experimental nanogels based on polymeric nanoparticles (NPs) and zinc, calcium or doxycycline-loaded NPs were applied to citric acid etched dentine to facilitate the occlusion of tubules and the mineralization of the dentine surface. Dentine surfaces were studied by X-ray diffraction and transmission electron microscopy through selected area diffraction and bright-field imaging. RESULTS: Crystals at the dentine surface were identified as hydroxyapatite with the highest crystallographic maturity and crystallite size in dentine treated with Zn-NPs-based gel. Texture increased in all samples from 24 h to 7 days, except in dentine surfaces treated with Zn-NPs gel. Polyhedral, plate-like and drop-like shaped apatite crystals constituted the bulk of minerals in dentine treated with Zn-NPs gel, after 7 days. Polymorphic, cubic and needle-like shaped crystals distinguished minerals, with more amorphous characteristics in dentine treated with Ca-NPs gel after 7 days than that found when Zn-NPs were applied. Doxycycline-NPs produced the smallest crystallites with poor crystallinity, maturity and chemical stability. CONCLUSIONS: Crystalline and amorphous phases of newly formed hydroxyapatite were described in both types of dentine treated with Zn-NPs as well as Ca-NPs gels with multiple shapes of crystallites. Crystal shapes ranged from rounded/drop-like or plate-like crystals to needle-like or polyhedral and cubic apatite appearance.

Antimicrobial effect of nanostructured membranes for guided tissue regeneration: an in vitro study.

OBJECTIVE: The purpose of this in vitro study was to evaluate the antibacterial effect of a novel non-resorbable, bioactive polymeric nanostructured membrane (NMs), when doped with zinc, calcium and doxycycline. METHODS: A validated in vitro subgingival biofilm model with six bacterial species (Streptococcus oralis, Actinomyces naeslundii, Veillonela parvula, Fusobacterium nucleatum, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans) was used. The experimental NMs, with and without being doped with doxycycline, calcium and zinc, were placed on hydroxyapatite (HA) discs. As positive control membranes, commercially available dense polytetrafluoroethylene (d-PTFE) membranes were used and, as negative controls, the HA discs without any membrane. The experimental, positive and negative control discs were exposed to a mixed bacterial suspension, at 37 °C under anaerobic conditions, during 12, 24, 48 and 72 h. The resulting biofilms were analyzed through scanning electron microscopy (SEM), to study their structure, and by quantitative polymerase chain reaction (qPCR), to assess the bacterial load, expressed as colony forming units (CFU) per mL. Differences between experimental and control groups were evaluated with the general linear model and the Bonferroni adjustment. RESULTS: As shown by SEM, all membrane groups, except the NMs with doxycycline, resulted in structured biofilms from 12-72 hours. Similarly, only the membranes loaded with doxycycline demonstrated a significant reduction in bacterial load during biofilm development, when compared with the control groups (p < 0.001). SIGNIFICANCE: Doxycycline-doped nanostructured membranes have an impact on biofilm growth dynamics by significant reducing the bacterial load.

Antibacterial effects of polymeric PolymP-n Active nanoparticles. An in vitro biofilm study.

OBJECTIVE: to study the antibacterial effect of polymeric PolymP-n Active nanoparticles using an in vitro subgingival biofilm model. METHODS: Hydroxyapatite discs coated with five modalities of nanoparticles (NPs): NPs, NPs doped with zinc, calcium, silver and doxycycline, PBS as control, and Streptococcus oralis, Actinomyces naeslundii, Veillonella parvula, Fusobacterium nucleatum, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were studied in a static in vitro biofilm model (12, 24, 48, and 72h). Nano-roughness of the different disc surfaces (SRa, in nm) and morphological characteristic of the biofilms (thickness (µm) and bacterial viability) were studied by different microscopy modalities. Quantitative Polymerase Chain Reaction was used to assess the effect of the nanoparticles on the bacterial load (colony forming unit per milliliter) (CFUmL-1). Analysis of variance and post-hoc testing with T3 Dunnett́s, and Student Newman Keuls correction was used. Results were considered statistically significant at p<0.05. RESULTS: Surfaces containing the different nanoparticles showed significant increments in roughness when compared to controls (p<0.05). A similar biofilm formation and dynamics was observed, although reductions in bacterial viability were detected in biofilms in contact with the different nanoparticles, more pronounced with silver and doxycycline NPs. Doxycycline-NPs biofilms resulted in unstructured biofilm formation and significantly lower number of the six species when compared with the other nanoparticles specimens and controls (p<0.001 in all cases). SIGNIFICANCE: Polymeric PolymP-n Active nanoparticles when combined with silver and doxycycline showed a significant antibacterial effect when tested in an in vitro subgingival biofilm model.

A rapid, sensitive screening test for polycyclic aromatic hydrocarbons applied to Antarctic water.

We describe a rapid, sensitive, fluorescence screening test for polycyclic aromatic hydrocarbons in water samples that avoids more costly time-consuming methods. The screening test works by detecting benzo[a]pyrene. It runs without the need for any pre-concentration step, thus rendering it suitable for routine use in water-quality-control laboratories. The test recognizes contaminated samples rapidly (150 s) and inexpensively with a cut-off level of 10 ng l(-1), which is the value that the European Union and World Health Organization (WHO) have laid down in its assessment of the quality of water for human consumption. This was first ascertained by analysing tap and waste-water samples before studying environmental water samples from the Antarctic region. The reliability of the screening test was 2% false positives and 4% false negatives in 200 samples of tap and waste-water. The applicability was confirmed by the fact that the predictions of the screening test coincided exactly with results obtained with gas chromatography-mass spectrometry assays. We also discuss the polluted Antarctic samples and the possible sources of the contamination involved.

Polymer nanocarriers for dentin adhesion.

To obtain more durable adhesion to dentin, and to protect collagen fibrils of the dentin matrix from degradation, calcium- and phosphate-releasing particles have been incorporated into the dental adhesive procedure. The aim of the present study was to incorporate zinc-loaded polymeric nanocarriers into a dental adhesive system to facilitate inhibition of matrix metalloproteinases (MMPs)-mediated collagen degradation and to provide calcium ions for mineral deposition within the resin-dentin bonded interface. PolymP- N : Active nanoparticles (nanoMyP) were zinc-loaded through 30-minute ZnCl2 immersion and tested for bioactivity by means of 7 days' immersion in simulated body fluid solution (the Kokubo test). Zinc-loading and calcium phosphate depositions were examined by scanning and transmission electron microscopy, elemental analysis, and x-ray diffraction. Nanoparticles in ethanol solution infiltrated into phosphoric-acid-etched human dentin and Single Bond (3M/ESPE) were applied to determine whether the nanoparticles interfered with bonding. Debonded sticks were analyzed by scanning electron microscopy. A metalloproteinase collagen degradation assay was also performed in resin-infiltrated dentin with and without nanoparticles, measuring C-terminal telopeptide of type I collagen (ICTP) concentration in supernatants, after 4 wk of immersion in artificial saliva. Numerical data were analyzed by analysis of variance (ANOVA) and Student-Newman-Keuls multiple comparisons tests (p < .05). Nanoparticles were effectively zinc-loaded and were shown to have a chelating effect, retaining calcium regardless of zinc incorporation. Nanoparticles failed to infiltrate demineralized intertubular dentin and remained on top of the hybrid layer, without altering bond strength. Calcium and phosphorus were found covering nanoparticles at the hybrid layer, after 24 h. Nanoparticle application in etched dentin also reduced MMP-mediated collagen degradation. Tested nanoparticles may be incorporated into dental adhesive systems to provide the appropriate environment in which dentin MMP collagen degradation is inhibited and mineral growth can occur.

Synthesis and characterization of a molecularly imprinted polymer optosensor for TEXs-screening in drinking water.

A molecularly imprinted polymer (MIP) was synthesized using toluene as template, and was implemented in a fluorescence optosensor (λ(exc)=260 nm, λ(em)=284 nm) for the screening of TEXs (toluene, ethylbenzene and xylenes) in drinking water. All the parameters which can affect the sensitivity and selectivity of the optical sensing phase, were carefully optimized. The screening test runs without the need for any pre-concentration step, thus rendering it suitable for routine use in water-quality-control laboratories. The test recognizes contaminated samples rapidly (81 s) and inexpensively with a cut-off level of 700 µg L(-1) ethylbenzene which corresponds with the maximum contaminant level (MCL) established by US Environmental Protection Agency (EPA) in drinking water. The threshold value of the screening test for the cut-off level was 8.27±0.57 a.u. (95% confidence level, n=10). The reliability of the screening test was 32% false positives and 0% false negatives for 50 samples, and its applicability has been demonstrated by analyzing 15 samples of mineral, tap and river waters obtaining 0% false negatives.