Novel potential scaffold for periodontal tissue engineering.

OBJECTIVE: The objective of the study is characterization of novel calcium and zinc-loaded electrospun matrices to be used for periodontal regeneration. MATERIALS AND METHODS: A polymethylmetacrylate-based membrane was calcium or zinc loaded. Matrices were characterized morphologically by atomic force and scanning electron microscopy and mechanically probed by a nanoindenter. Biomimetic calcium phosphate precipitation on polymeric tissues was assessed. Cell viability tests were performed using oral mucosa fibroblasts. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests or by ANOVA and Student-Newman-Keuls multiple comparisons. RESULTS: Zinc and calcium loading on matrices did not modify their morphology but increased nanomechanical properties and decreased nanoroughness. Precipitation of calcium and phosphate on the matrix surfaces was observed in zinc-loaded specimens. Matrices were found to be non-toxic to cells in all the assays. Calcium- and zinc-loaded scaffolds presented a very low cytotoxic effect. CONCLUSIONS: Zinc-loaded membranes permit cell viability and promoted mineral precipitation in physiological conditions. Based on the tested nanomechanical properties and scaffold architecture, the proposed membranes may be suitable for cell proliferation. CLINICAL RELEVANCE: The ability of zinc-loaded matrices to promote precipitation of calcium phosphate deposits, together with their observed non-toxicity and its surface chemistry allowing covalent binding of proteins, may offer new strategies for periodontal regeneration.

Zinc-modified nanopolymers improve the quality of resin-dentin bonded interfaces.

INTRODUCTION: Demineralized collagen fibers at the hybrid layer are susceptible to degradation. Remineralization may aid to improve bond longevity. OBJECTIVES: The aim of the present study was to infiltrate zinc and calcium-loaded polymeric nanoparticles into demineralized dentin to facilitate hybrid layer remineralization. MATERIALS AND METHODS: Zinc or calcium-loaded polymeric nanoparticles were infiltrated into etched dentin, and Single Bond Adhesive was applied. Bond strength was tested after 24 h and 6 months storage. Nanomechanical properties, dye-assisted confocal laser microscopy, and Masson's trichrome staining evaluation were performed to assess for the hybrid layer morphology, permeability, and remineralization ability after 24 h and 3 months. Data were analyzed by ANOVA and Student-Newman-Keuls multiple comparisons tests (p < 0.05). RESULTS: Immediate bond strength was not affected by nanoparticles infiltration (25 to 30 MPa), while after 6 months, bond strengths were maintained (22 to 24 MPa). After 3 months, permeability occurred only in specimens in which nanoparticles were not infiltrated. Dentin remineralization, at the bottom of the hybrid layer, was observed in all groups. After microscopy analysis, zinc-loaded nanoparticles were shown to facilitate calcium deposition throughout the entire hybrid layer. Young's modulus at the hybrid layer increased from 2.09 to 3.25 GPa after 3 months, in specimens with zinc nanoparticles; meanwhile, these values were reduced from 1.66 to 0.49 GPa, in the control group. CONCLUSION: Infiltration of polymeric nanoparticles into demineralized dentin increased long-term bond strengths. Zinc-loaded nanoparticles facilitate dentin remineralization within the complete resin-dentin interface. CLINICAL RELEVANCE: Resin-dentin bond longevity and dentin remineralization at the hybrid layer were facilitated by zinc-loaded nanoparticles.

Highly deformable and strongly magnetic semi-interpenetrating hydrogels based on alginate or cellulose.

The effective implementation of many of the applications of magnetic hydrogels requires the development of innovative systems capable of withstanding a substantial load of magnetic particles to ensure exceptional responsiveness, without compromising their reliability and stability. To address this challenge, double-network hydrogels have emerged as a promising foundation, thanks to their extraordinary mechanical deformability and toughness. Here, we report a semi-interpenetrating polymer networks (SIPNs) approach to create diverse magnetic SIPNs hydrogels based on alginate or cellulose, exhibiting remarkable deformability under certain stresses. Achieving strong responsiveness to magnetic fields is a key objective, and this characteristic is realized by the incorporation of highly magnetic iron microparticles at moderately large concentrations into the polymer network. Remarkably, the SIPNs hydrogels developed in this research accommodate high loadings of magnetic particles without significantly compromising their physical properties. This feature is essential for their use in applications that demand robust responsiveness to applied magnetic fields and overall stability, such as a hydrogel luminescent oxygen sensor controlled by magnetic fields that we designed and tested as proof-of-concept. These findings underscore the potential and versatility of magnetic SIPNs hydrogels based on carbohydrate biopolymers as fundamental components in driving the progress of advanced hydrogels for diverse practical implementations.

Antibacterial Effect of Functionalized Polymeric Nanoparticles on Titanium Surfaces Using an In Vitro Subgingival Biofilm Model.

This investigation aimed to evaluate the antibacterial effect of polymeric nanoparticles (NPs), functionalized with calcium, zinc, or doxycycline, using a subgingival biofilm model of six bacterial species (Streptococcus oralis,Actinomyces naeslundii, Veillonela parvula, Fusobacterium nucleatum, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans) on sandblasted, large grit, acid-etched titanium discs (TiDs). Undoped NPs (Un-NPs) or doped NPs with calcium (Ca-NPs), zinc (Zn-NPs), or doxycycline (Dox-NPs) were applied onto the TiD surfaces. Uncovered TiDs were used as negative controls. Discs were incubated under anaerobic conditions for 12, 24, 48, and 72 h. The obtained biofilm structure was studied by scanning electron microscopy (SEM) and its vitality and thickness by confocal laser scanning microscopy (CLSM). Quantitative polymerase chain reaction of samples was used to evaluate the bacterial load. Data were evaluated by analysis of variance (p < 0.05) and post hoc comparisons with Bonferroni adjustments (p < 0.01). As compared with uncovered TiDs, Dox-NPs induced higher biofilm mortality (47.21% and 85.87%, respectively) and reduced the bacterial load of the tested species, after 72 h. With SEM, scarce biofilm formation was observed in Dox-NPs TiDs. In summary, Dox-NPs on TiD reduced biofilm vitality, bacterial load, and altered biofilm formation dynamics.

Novel Pastes Containing Polymeric Nanoparticles for Dentin Hypersensitivity Treatment: An In Vitro Study.

Tubule occlusion and remineralization are considered the two main goals of dentin hypersensitivity treatment. The objective is to assess the ability of dentifrices containing zinc-doped polymeric nanoparticles (NPs) to enduringly occlude the dentinal tubules, reinforcing dentin's mechanical properties. Fifteen dentin surfaces were acid-treated for dentinal tubule exposure and brushed with (1) distilled water, or with experimental pastes containing (2) 1% of zinc-doped NPs, (3) 5% of zinc-doped NPs, (4) 10% of zinc-doped NPs or (5) Sensodyne®. Topographical and nanomechanical analyses were performed on treated dentin surfaces and after a citric acid challenge. ANOVA and Student-Newman-Keuls tests were used (p < 0.05). The main results indicate that all pastes produced tubule occlusion (100%) and reinforced mechanical properties of intertubular dentin (complex modulus was above 75 GPa). After the citric acid challenge, only those pastes containing zinc-doped NPs were able to maintain tubular occlusion, as specimens treated with Sensodyne® have around 30% of tubules opened. Mechanical properties were maintained for dentin treated with Zn-doped NPs, but in the case of specimens treated with Sensodyne®, complex modulus values were reduced below 50 GPa. It may be concluded that zinc-doped NPs at the lowest tested concentration produced acid-resistant tubular occlusion and increased the mechanical properties of dentin.

Melatonin-doped polymeric nanoparticles induce high crystalline apatite formation in root dentin.

OBJECTIVE: To investigate the effect of novel polymeric nanoparticles (NPs) doped with melatonin (ML) on nano-hardness, crystallinity and ultrastructure of the formed hydroxyapatite after endodontic treatment. METHODS: Undoped-NPs and ML-doped NPs (ML-NPs) were tested at radicular dentin, after 24 h and 6 m. A control group without NPs was included. Radicular cervical and apical dentin surfaces were studied by nano-hardness measurements, X-ray diffraction and transmission electron microscopy. Mean and standard deviation were analyzed by ANOVA and Student-Newman-Keuls multiple comparisons (p < 0.05). RESULTS: Cervical dentin treated with undoped NPs maintained its nano-hardness values after 6 m of storage being [24 h: 0.29 (0.01); 6 m: 0.30 (0.02) GPa], but it decreased at apical dentin [24 h: 0.36 (0.01); 6 m: 0.28 (0.02) GPa]. When ML-NPs were used, nano-hardness was similar over time [24h: 0.31 (0.02); 6 m: 0.28 (0.03) GPa], at apical dentin. Root dentin treated with ML-NPs produced, in general, high crystallinity of new minerals and thicker crystals than those produced in the rest of the groups. After 6 m, crystals became organized in randomly oriented polyhedral, square polygonal block-like apatite or drop-like apatite polycrystalline lattices when ML-NPs were used. Undoped NPs generated poor crystallinity, with preferred orientation of small crystallite and increased microstrain. SIGNIFICANCE: New polycrystalline formations encountered in dentin treated with ML-NPs may produce structural dentin stability and high mechanical performance at the root. The decrease of mechanical properties over time in dentin treated without NPs indicates scarce remineralization potential, dentin demineralization and further potential degradation. The amorphous stage may provide high hydroxyapatite solubility and remineralizing activity.

Melatonin-doped polymeric nanoparticles reinforce and remineralize radicular dentin: Morpho-histological, chemical and biomechanical studies.

OBJECTIVES: To investigate the effectiveness of novel polymeric nanoparticles (NPs) doped with melatonin (ML) in reducing dentin permeability and facilitating dentin remineralization after endodontic treatment. METHODS: The effect of undoped NPs and ML-doped NPs (ML-NPs) was tested in radicular dentin, at 24 h and 6 m. A control group without NPs was included. ML liberation was measured. Radicular dentin was assessed for fluid filtration. Dentin remineralization was analyzed by scanning electron microscopy, AFM, Young's modulus (Ei), Nano DMA-tan delta, and Raman analysis. RESULTS: ML release ranged from 1.85 mg/mL at 24 h to 0.033 mg/mL at 28 d. Both undoped NPs and ML-NPs treated dentin exhibited the lowest microleakage, but samples treated with ML-NPs exhibited hermetically sealed dentinal tubules and extended mineral deposits onto dentin. ML-NPs promoted higher and durable Ei, and functional remineralization at root dentin, generating differences between the values of tan delta among groups and creating zones of stress concentration. Undoped-NPs produced closure of some tubules and porosities at the expense of a relative mineral amorphization. Chemical remineralization based on mineral and organic assessments was higher in samples treated with ML-NPs. When using undoped NPs, precipitation of minerals occurred; however, radicular dentin was not mechanically reinforced but weakened over time. SIGNIFICANCE: Application of ML-NPs in endodontically treated teeth, previous to the canal filling step, is encouraged due to occlusion of dentinal tubules and the reinforcement of the radicular dentin structure.

Polymeric nanoparticles for endodontic therapy.

The effectiveness of novel polymeric nanoparticles (NPs) application in reducing dentin permeability and facilitating dentin remineralization after endodontic treatment was evaluated. The effect of undoped NPs, zinc, calcium and doxycycline-doped NPs (Zn-NPs, Ca-NPs and D-NPs, respectively) was tested in radicular dentin. A control group without NPs was included. Radicular dentin was assessed for fluid filtration. Dentin remineralization was analyzed by scanning and transmission electron microscopy, energy-dispersive analysis, AFM, Young's modulus (Ei), Nano DMA, Raman, and X-Ray Diffraction analysis. Ca-NPs and Zn-NPs treated dentin exhibited the lowest microleakage with hermetically sealed dentinal tubules and a zinc-based salt generation onto dentin. Zn-NPs favored crystallinity and promoted the highest Ei and functional remineralization at the apical dentin, generating differences between the values of complex modulus among groups. Ca-NPs produced closure of tubules and porosities at the expense of a relative mineral amorphization, without creating zones of stress concentration. The highest sealing efficacy was obtained in Zn-NPs-treated samples, along with the highest values of Young's modulus and dentin mineralization. These high values of Ei were obtained by closing voids, cracks, pores and tubules, and by strengthening the root dentin. When using undoped NPs or Ca-NPs, deposition of minerals occurred, but radicular dentin was not mechanically reinforced. Therefore, application of Zn-NPs in endodontically treated teeth previous to the canal filling is encouraged.

Modified Polymeric Nanoparticles Exert In Vitro Antimicrobial Activity Against Oral Bacteria.

Polymeric nanoparticles were modified to exert antimicrobial activity against oral bacteria. Nanoparticles were loaded with calcium, zinc and doxycycline. Ions and doxycycline release were measured by inductively coupled plasma optical emission spectrometer and high performance liquid chromatography. Porphyromonas gingivalis, Lactobacillus lactis, Streptoccocus mutans, gordonii and sobrinus were grown and the number of bacteria was determined by optical density. Nanoparticles were suspended in phosphate-buffered saline (PBS) at 10, 1 and 0.1 mg/mL and incubated with 1.0 mL of each bacterial suspension for 3, 12, and 24 h. The bacterial viability was assessed by determining their ability to cleave the tetrazolium salt to a formazan dye. Data were analyzed by ANOVA and Scheffe’s F (p < 0.05). Doxycycline doping efficacy was 70%. A burst liberation effect was produced during the first 7 days. After 21 days, a sustained release above 6 µg/mL, was observed. Calcium and zinc liberation were about 1 and 0.02 µg/mL respectively. The most effective antibacterial material was found to be the Dox-Nanoparticles (60% to 99% reduction) followed by Ca-Nanoparticles or Zn-Nanoparticles (30% to 70% reduction) and finally the non-doped nanoparticles (7% to 35% reduction). P. gingivalis, S. mutans and L. lactis were the most susceptible bacteria, being S. gordonii and S. sobrinus the most resistant to the tested nanoparticles.

Effect of functionalized PHEMA micro- and nano-particles on the viscoelastic properties of fibrin-agarose biomaterials.

Two types of PHEMA-based particles, exhibiting either carboxyl or tertiary ammine functional groups, were incorporated to fibrin-agarose (FA) hydrogels, and the effect of the addition of these synthetic particles on the viscoelastic and microstructural properties of the biomaterials was evaluated. Experimental results indicated that the incorporation of both types of polymeric particles to FA scaffolds was able to improve the biomechanical properties of the biomaterials under steady state and oscillatory shear stresses, resulting in scaffolds characterized by higher values of the storage, loss, and shear moduli. In addition, the microstructural evaluation of the scaffolds showed that the nanoparticles exhibiting carboxyl functional groups were homogeneously distributed across the fibrous network of the hydrogels. The addition of both types of artificial polymeric particles was able to enhance the viscoelastic properties of the FA hydrogels, allowing the biomaterials to reach levels of mechanical consistency under shear stresses in the same range of some human native soft tissues, which could allow these biomaterials to be used as scaffolds for new tissue engineering applications. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 738-745, 2018.

Bioactive Polymeric Nanoparticles for Periodontal Therapy.

AIMS: to design calcium and zinc-loaded bioactive and cytocompatible nanoparticles for the treatment of periodontal disease. METHODS: PolymP-nActive nanoparticles were zinc or calcium loaded. Biomimetic calcium phosphate precipitation on polymeric particles was assessed after 7 days immersion in simulated body fluid, by scanning electron microscopy attached to an energy dispersive analysis system. Amorphous mineral deposition was probed by X-ray diffraction. Cell viability analysis was performed using oral mucosa fibroblasts by: 1) quantifying the liberated deoxyribonucleic acid from dead cells, 2) detecting the amount of lactate dehydrogenase enzyme released by cells with damaged membranes, and 3) by examining the cytoplasmic esterase function and cell membranes integrity with a fluorescence-based method using the Live/Dead commercial kit. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests. RESULTS: Precipitation of calcium and phosphate on the nanoparticles surfaces was observed in calcium-loaded nanoparticles. Non-loaded nanoparticles were found to be non-toxic in all the assays, calcium and zinc-loaded particles presented a dose dependent but very low cytotoxic effect. CONCLUSIONS: The ability of calcium-loaded nanoparticles to promote precipitation of calcium phosphate deposits, together with their observed non-toxicity may offer new strategies for periodontal disease treatment.

Synthesis of a novel polyurethane-based-magnetic imprinted polymer for the selective optical detection of 1-naphthylamine in drinking water.

The first polyurethane based magnetic-MIP for the selective detection of 1-naphthylamine (1-NA) in drinking water has been synthesised. The synthesis has been carried out in a two-step process: first,the incorporation of magnetite-coated-oleic acid nanoparticles (-Fe3O4-OA) into a lipophilic polymeric matrix (poly-MMA-co-EDMA) and second, the encapsulation of these magnetic seeds into the MIP structure by precipitation polymerisation. The mag-MIP was first RHTEM imaged showing a well-organised material with magnetite within the material and the imprinted polymer coating the magnetic core. Thereafter,it was evaluated by batch rebinding analysis and the derived Freundlich isotherm, calculating the number of binding sites (N(K(min)-K(max))=2.63 and 0.79 mmol g⁻¹, for mag-MIP and mag-NIP, respectively)and apparent average adsorption constant (K(K(min)-K(max))=3.31 and 3.06 mmol⁻¹, for mag-MIP and mag-NIP, respectively) showing a very effective imprinting process.We have also developed a magnetic optical sensor MIP by using an optical fiber coupled with a magnetic separator. An unexpected selectivity for 1-NA was revealed allowing the detection of this molecule in water, even in the presence of 4 structurally related compounds (2-naphthylamine, 1-naphthol, 2-naphthol and 1-naphthalenemethylamine), with a low limit of detection (LOD) = 18 ng mL⁻¹. Finally, we applied this new hybrid material to the analysis of 1-NA in tap and mineral waters, obtaining a 91.6%average recovery rate.

A semi-empirical model to simplify the synthesis of homogeneous and transparent cross-linked polymers and their application in the preparation of optical sensing films.

We propose a simple, semi-empirical model based on Hansen's solubility parameters for simplifying the synthesis and the optimization of homogeneous and transparent cross-linked polymers in order to obtain optical sensing films. More than 740 experiments were undertaken to demonstrate the reliability of the model and several applications are proposed. We have demonstrated that our model can help in the synthesis and optimization (percentage of cross-linker, changes in hydrophilicity, selection of porogens, quantity of template etc.) of homogeneous and transparent MIPs and NIPs (molecularly imprinted polymers) with VOCs; after the synthesis of 440 polymers in the homogeneity zone only 4.32% of them (19 samples out of 440) were heterogeneous. We suggest a role for its use in the development of novel polymeric resins for detecting volatile organic compounds in water by measuring intrinsic fluorescence, in simplifying the synthesis of imprinted polymers and in decreasing the number of experiments required to optimize optical sensing membranes. In addition, it might also be used for synthesizing and optimizing MIPs with a non-volatile template.

Engineering of efficient phosphorescent iridium cationic complex for developing oxygen-sensitive polymeric and nanostructured films.

In this study, a novel phosphorescent Ir(III) complex [Ir(2-phenylpyridine)2(4,4'-bis(2-(4-N,N-methylhexylaminophenyl)ethyl)-2-2'-bipyridine)Cl] (for convenience, the complex was given the synonym N-948) has been designed and synthesized, to be used as an oxygen probe. It was characterized by spectroscopic and analytical methods when incorporated in a polystyrene and nanostructured metal oxide support. N-948 is the first Ir complex in the literature with a luminescence emission at a wavelength higher than 650 nm (665 nm), with a quantum yield higher than 0.50 (0.58 +/- 0.05) and an extremely long phosphorescence lifetime (102 micros) which has been used for developing oxygen-sensitive films. In addition, the new complex shows a Stern-Volmer constant which is 20 times higher than that of other Ir complexes known from the literature when they are immobilized in polystyrene. The sensing film shows long-term stability (up to 12 months), complete reversibility of the signal quenched by oxygen and a quick response time to various oxygen concentrations (<2 s changing from 10 vol% pO2 to 90 vol% pO2). Thus, it is an interesting and promising complex for developing oxygen-selective sensors for gas analysis and the analysis of dissolved oxygen.