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1.
Analyst ; 149(8): 2480, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38498079

RESUMO

Correction for 'An optical sensor for the sensitive determination of formaldehyde gas based on chromotropic acid and 4-aminoazobenzene immobilized in a hydrophilic membrane' by M. D. Fernández-Ramos et al., Analyst, 2023, 148, 4533-4538, https://doi.org/10.1039/D3AN01056B.

2.
Int J Biol Macromol ; 260(Pt 1): 129368, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38219926

RESUMO

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.


Assuntos
Celulose , Hidrogéis , Alginatos , Reprodutibilidade dos Testes , Polímeros , Fenômenos Magnéticos
3.
Vaccines (Basel) ; 11(11)2023 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-38005967

RESUMO

Comirnaty™ and Spikevax™ were the first vaccines approved for human use based on modified non-replicating mRNA lipophilic nanoparticle (mRNA-LNP) technology, with great success in the treatment of COVID-19. They have been used massively worldwide. One of the major inconveniences of these vaccines is related to pharmaceutical stability issues. Proper transportation, storage, and in-use handling before administration to patients are critical steps since failures can potentially reduce potency. In this research, the in-use stability of Comirnaty™ and Spikevax™ clinical samples was analysed and the results were compared. As changes in the size of the mRNA-LNPs are related to potency, these modifications were analysed by qualitative Dynamic Light Scattering (DLS) as a stability-indicating method for control and stressed vaccine samples. Strong stress factors (accelerated light irradiation, manual shaking, and vortex vibration) and conditions that mimic in-use handling (exposure to natural light and room temperature, repeated cycles of injections, and 24 h storage in syringes) were checked. The morphology of the mRNA-LNPs was analysed by Transmission Electron Microscopy (TEM) to better interpret and support the DLS results. Although the two vaccines are based on the same mRNA-LNP technology, the results demonstrate that they are characterised by very different particle size profiles and behaviours against different handling/stress conditions.

4.
Analyst ; 148(18): 4533-4538, 2023 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-37585267

RESUMO

Formaldehyde is a common contaminant in occupational and environmental atmospheres, prolonged exposure leads to health risks, and its determination is necessary to protect health. There is a great demand for portable, rapid, and sensitive methods that can be used in resource-limited settings. In this respect, a colorimetric sensor has been developed based on the colour change from pink to purple of co-immobilized chromotropic acid and 4-aminoazobenzene in hydroxypropyl methylcellulose when it is exposed to different concentrations of formaldehyde. The concentration of formaldehyde in the gas phase was quantified by measuring the change of the appropriate colour coordinates in response to the concentration of formaldehyde. A calibration curve was obtained for formaldehyde, with a useful concentration range from 0.08 to 0.6 ppmv. The detection limit was 0.016 ppmv, which is lower than the maximum exposure concentrations recommended by both the World Health Organization (WHO) and the Occupational Safety and Health Administration (OSHA). The optical sensor was found to have good reproducibility, with a relative standard deviation of 2.3 and 1.7% at 0.08 and 0.25 ppmv, respectively. The sensor can operate at room temperature and environmental humidity, 25 °C, and 50% RH, respectively. In addition, a study of interferents (acetaldehyde, toluene, methanol, ethyl acetate, acetone, acetic acid, carbon dioxide and ammonia) showed high selectivity for formaldehyde, which indicates that this membrane is a simple, fast, and economical alternative for quantifying the concentration of formaldehyde in different environments.


Assuntos
Colorimetria , Formaldeído , Reprodutibilidade dos Testes , Colorimetria/métodos , Naftalenossulfonatos
5.
ACS Appl Polym Mater ; 4(8): 6054-6066, 2022 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-35991305

RESUMO

The high porosity, interconnected pore structure, and high surface area-to-volume ratio make the hydrophilic nonwoven nanofiber membranes (NV-NF-Ms) promising nanostructured supports for enzyme immobilization in different biotechnological applications. In this work, NV-NF-Ms with excellent mechanical and chemical properties were designed and fabricated by electrospinning in one step without using additives or complicated crosslinking processes after electrospinning. To do so, two types of ultrahigh-molecular-weight linear copolymers with very different mechanical properties were used. Methyl methacrylate-co-hydroxyethyl methacrylate (p(MMA)-co-p(HEMA)) and methyl acrylate-co-hydroxyethyl acrylate (p(MA)-co-p(HEA)) were designed and synthesized by reverse atom transfer radical polymerization (reverse-ATRP) and copper-mediated living radical polymerization (Cu0-MC-LRP), respectively. The copolymers were characterized by nuclear magnetic resonance (1H-NMR) spectroscopy and by triple detection gel permeation chromatography (GPC). The polarity, topology, and molecular weight of the copolymers were perfectly adjusted. The polymeric blend formed by (MMA)1002-co-(HEMA)1002 (M w = 230,855 ± 7418 Da; M n = 115,748 ± 35,567 Da; PDI = 2.00) and (MA)11709-co-(HEA)7806 (M w = 1.972 × 106 ± 33,729 Da; M n = 1.395 × 106 ± 35,019 Da; PDI = 1.41) was used to manufacture (without additives or chemical crosslinking processes) hydroxylated nonwoven nanofiber membranes (NV-NF-Ms-OH; 300 nm in fiber diameter) with excellent mechanical and chemical properties. The morphology of NV-NF-Ms-OH was studied by scanning electron microscopy (SEM). The suitability for enzyme binding was proven by designing a palette of different surface functionalization to enable both reversible and irreversible enzyme immobilization. NV-NF-Ms-OH were successfully functionalized with vinyl sulfone (281 ± 20 µmol/g), carboxyl (560 ± 50 µmol/g), and amine groups (281 ± 20 µmol/g) and applied for the immobilization of two enzymes of biotechnological interest. Galactose oxidase was immobilized on vinyl sulfone-activated materials and carboxyl-activated materials, while laccase was immobilized onto amine-activated materials. These preliminary results are a promising basis for the application of nonwoven membranes in enzyme technology.

6.
Polymers (Basel) ; 14(3)2022 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-35160348

RESUMO

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.

7.
Nanomaterials (Basel) ; 11(11)2021 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-34835914

RESUMO

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.

8.
Dent Mater ; 37(11): 1698-1713, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34544591

RESUMO

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.


Assuntos
Melatonina , Nanopartículas , Apatitas , Dentina , Humanos , Polímeros
9.
Polymers (Basel) ; 13(7)2021 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-33800569

RESUMO

Polymeric membranes are employed in guided bone regeneration (GBR) as physical barriers to facilitate bone in-growth. A bioactive and biomimetic membrane with the ability to participate in the healing and regeneration of the bone is necessary. The aim of the present study was to analyze how novel silicon dioxide composite membranes functionalized with zinc or doxycycline can modulate the osteoblasts' proliferation, differentiation, and expression of selected antigenic markers related to immunomodulation. Nanostructured acrylate-based membranes were developed, blended with silica, and functionalized with zinc or doxycycline. They were subjected to MG63 osteoblast-like cells culturing. Proliferation was assessed by MTT-assay, differentiation by evaluating the alkaline phosphatase activity by a spectrophotometric method and antigenic phenotype was assessed by flow cytometry for selected markers. Mean comparisons were conducted by one-way ANOVA and Tukey tests (p < 0.05). The blending of silica nanoparticles in the tested non-resorbable polymeric scaffold improved the proliferation and differentiation of osteoblasts, but doxycycline doped scaffolds attained the best results. Osteoblasts cultured on doxycycline functionalized membranes presented higher expression of CD54, CD80, CD86, and HLA-DR, indicating a beneficial immunomodulation activity. Doxycycline doped membranes may be a potential candidate for use in GBR procedures in several challenging pathologies, including periodontal disease.

10.
J Dent ; 109: 103676, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33891990

RESUMO

OBJECTIVES: To analyze how novel developed silicon dioxide composite membranes, functionalized with zinc or doxycycline, can modulate the expression of genes related to the osteogenic functional capacity of osteoblastic cells. METHODS: The composite nanofibers membranes were manufactured by using a novel polymeric blend and 20 nm silicon dioxide nanoparticles (SiO2-NPs). To manufacture the membranes, 20 nm SiO2-NPs were added to the polymer solution and the resulting suspension was processed by electrospinning. In a second step, the membranes were functionalized with zinc or doxycycline. Then, they were subjected to MG63 osteoblast-like cells culturing for 48 h. After this time, real-time quantitative polymerase chain reaction (RT-qPCR) was carried out to study the expression of Runx-2, OSX, ALP, OSC, OPG, RANKL, Col-I, BMP-2, BMP-7, TGF-ß1, VEGF, TGF-ßR1, TGF- ßR2, and TGF-ßR3. Mean comparisons were conducted by One-way ANOVA and Tukey tests (p < 0.05). RESULTS: In general, the blending of SiO2-NPs in the tested non-resorbable polymeric scaffold improves the expression of osteogenic genes over the control membranes. Doxycycline doping of experimental scaffolds attained the best results, encountering up-regulation of BMP-2, ALP, OPG, TGFß-1 and TGFß-R1. Membranes with zinc induced a significant increase in the expression of Col-I, ALP and TGF ß1. Both, zinc and doxycycline functionalized membranes enormously down-regulated the expression of RANKL. CONCLUSIONS: Zinc and doxycycline doped membranes are bioactive inducing overexpression of several osteogenic gene markers. CLINICAL SIGNIFICANCE: Doxycycline doped membranes may be a potential candidate for use in GBR procedures in several challenging pathologies, including periodontal diseases.


Assuntos
Doxiciclina , Dióxido de Silício , Diferenciação Celular , Expressão Gênica , Osteoblastos , Osteogênese
11.
Dent Mater ; 37(7): 1107-1120, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33846017

RESUMO

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.


Assuntos
Melatonina , Nanopartículas , Dentina , Microscopia Eletrônica de Varredura , Polímeros , Raiz Dentária
12.
J Mech Behav Biomed Mater ; 103: 103606, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32090933

RESUMO

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.


Assuntos
Dentina , Nanopartículas , Microscopia Eletrônica de Varredura , Polímeros , Raiz Dentária , Zinco
13.
Talanta ; 187: 83-90, 2018 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-29853069

RESUMO

In this study, we have optimised the sterically directed attachment of biomolecules on the surface of coaxial membranes prepared by co-electrospinning which have been proved to be a material with very high performance for the development of biosensors with optical oxygen transduction. Uricase has been used as model enzyme. Two sterically directed strategies: a) covalent attachment via maleimide, and b) affinity bonding via biotin-streptavidin interaction, have been tested in order to preserve the enzymatic activity of uricase and to improve the analytical figures of merits on the determination of uric acid. The best results were obtained with biotin-streptavidin affinity interaction and using a biotinylation reagent containing a polyethylene glycol chain. The developed biosensor showed high sensitivity towards uric acid with a detection limit of 0.5 µM, a quantification limit of 1.8 µM and linear range from 1.8 to 250 µM. The applicability of the membrane as biosensor with optical oxygen transduction was proved by determining uric acid in serum samples. The obtained results showed a good correlation (0.999) with those obtained by an external reference laboratory.

14.
Materials (Basel) ; 11(6)2018 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-29904023

RESUMO

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.

15.
Anal Chim Acta ; 1015: 66-73, 2018 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-29530253

RESUMO

A multifunctional material based on co-electrospinning has been developed as a basic material for the development of biosensors with optical oxygen transduction. It is based on coaxial nanofibres: inner fibres containing an oxygen sensitive dye and outer fibres containing aldehyde groups to allow the formation of Schiff bases with the amino groups of the enzyme. The resulting material preserves the oxygen sensing properties of the inner optical transducer as well as exhibits a high capacity for immobilizing molecules on its surface. Uricase has been selected as model enzyme and several parameters (temperature, pH, reaction time, buffer, and enzyme concentration) have been optimised to demonstrate the versatility of this novel multifunctional material in the development of biosensors with optical oxygen transduction for determining uric acid in serum samples. It suggests that the proposed multifunctional material can provide a promising multifunctional platform for biosensing applications.


Assuntos
Técnicas Biossensoriais , Oxigênio/metabolismo , Urato Oxidase/metabolismo , Oxigênio/sangue , Espectrofotometria Ultravioleta , Propriedades de Superfície
16.
J Biomed Mater Res A ; 106(3): 738-745, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29052310

RESUMO

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.


Assuntos
Materiais Biocompatíveis/química , Elasticidade , Fibrina/química , Nanopartículas/química , Poli-Hidroxietil Metacrilato/química , Sefarose/química , Fenômenos Biomecânicos , Hidrogéis/química , Resistência ao Cisalhamento , Viscosidade
17.
Clin Oral Investig ; 21(9): 2695-2707, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28214952

RESUMO

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.


Assuntos
Materiais Biocompatíveis/farmacologia , Materiais Biomiméticos/farmacologia , Fosfatos de Cálcio/farmacologia , Fibroblastos/citologia , Mucosa Bucal/citologia , Engenharia Tecidual/instrumentação , Alicerces Teciduais , Zinco/farmacologia , Sobrevivência Celular , Humanos , Microscopia de Força Atômica , Microscopia Eletrônica de Varredura , Polimetil Metacrilato/química
18.
PLoS One ; 11(11): e0166217, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27820866

RESUMO

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.


Assuntos
Nanopartículas/administração & dosagem , Nanopartículas/química , Doenças Periodontais/tratamento farmacológico , Polímeros/química , Materiais Biomiméticos/química , Cálcio/química , Fosfatos de Cálcio/química , Humanos , Teste de Materiais/métodos , Microscopia Eletrônica de Varredura/métodos , Mucosa Bucal/efeitos dos fármacos , Propriedades de Superfície , Difração de Raios X/métodos , Zinco/química
19.
Biosens Bioelectron ; 82: 217-23, 2016 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-27085954

RESUMO

This paper describes a novel biosensor which combines the use of nanotechnology (non-woven nanofibre mat) with Solid Surface-Room Temperature Phosphorescence (SS-RTP) measurement for the determination of serotonin in human serum. The developed biosensor is simple and can be directly applied in serum; only requires a simple clean-up protocol. Therefore it is the first time that serotonin is analysed directly in serum with a non-enzymatic technique. This new approach is based on the covalent immobilization of serotonin directly from serum on a functional nanofibre material (Tiss®-Link) with a preactivated surface for direct covalent immobilization of primary and secondary amines, and the subsequent measurement of serotonin phosphorescent emission from the solid surface. The phosphorescent detection allows avoiding the interference from any fluorescence emission or scattering light from any molecule present in the serum sample which can be also immobilised on the nanofibre material. The determination of serotonin with this SS-RTP sensor overcomes some limitations, such as large interference from the matrix and high cost and complexity of many of the methods widely used for serotonin analysis. The potential applicability of the sensor in the clinical diagnosis was demonstrated by analysing serum samples from seven healthy volunteers. The method was validated with an external reference laboratory, obtaining a correlation coefficient of 0.997 which indicates excellent correlation between the two methods.


Assuntos
Medições Luminescentes/métodos , Serotonina/sangue , Técnicas Biossensoriais/métodos , Humanos , Limite de Detecção , Nanofibras/química , Nanotecnologia , Temperatura
20.
Clin Oral Investig ; 20(9): 2411-2420, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26832781

RESUMO

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.


Assuntos
Adesivos Dentinários/química , Dentina/química , Nanopartículas/química , Cimentos de Resina/química , Remineralização Dentária , Zinco/química , Condicionamento Ácido do Dente , Cálcio/química , Colagem Dentária , Módulo de Elasticidade , Humanos , Teste de Materiais , Microscopia Confocal , Dente Serotino , Coloração e Rotulagem , Propriedades de Superfície , Resistência à Tração
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