HPLC quantitative determination of acrylic monomers released from resins used in dental restoration

Poster presented at the First International Congress of CiiEM - From Basic Sciences To Clinical Research. Egas Moniz, Caparica, Portugal, 27-28 November 2015

Versatility of polymethacrylate monoliths for chromatographic purification of biomolecules

Polymethacrylate monoliths, specifically poly(glycidyl methacrylate-co-ethylene dimethacrylate) or poly(GMA-co-EDMA) monoliths, are a new generation of chromatographic supports and are significantly different from conventional particle-based adsorbents, membranes, and other monolithic supports for biomolecule purification. Similar to other monoliths, polymethacrylate monoliths possess large pores which allow convective flow of mobile phase and result in high flow rates at reduced pressure drop, unlike particulate supports. The simplicity of the adsorbent synthesis, pH resistance, and the ease and flexibility of tailoring their pore size to that of the target biomolecule are the key properties which differentiate polymethacrylate monoliths from other monoliths. Polymethacrylate monoliths are endowed with reactive epoxy groups for easy functionalization (with anion-exchange, hydrophobic, and affinity ligands) and high ligand retention. In this review, the structure and performance of polymethacrylate monoliths for chromatographic purification of biomolecules are evaluated and compared to those of other supports. The development and use of polymethacrylate monoliths for research applications have grown rapidly in recent times and have enabled the achievement of high through-put biomolecule purification on semi-preparative and preparative scales.

Electrooptic proprieties of layer-by-layer films of a azobenzene containing copolymer

The methacrylic copolymer functionalized with the azo chromophore 4-[N-ethyl-N-(2-hydroxiethyl)]-amino-2′-chloro-4-nitroazobenzene (MMADR13), in its polyelectrolyte form, can be used to fabricate thin films by the layer-by-layer (LbL) technique just if one alternates this anionic polyelectrolyte with a cationic polyelectrolyte such as poly(allylamine hydrochloride) (PAH). Since PAH does not present any particular optical functionality, the main final film feature will came from the side chain DR13 azo-chromophore group due to its large nonlinear optical properties and photoisomerization capabilities. This work reports the electrooptic activity of MMADR13/DR13 LBL films, which arises from the high hiperpolarizability about the azo side chain group.

Synthesis and characterization of polymers containing oligothiophenes

In the present work a series of thiophene oligomers of three and six thiophene units were synthesized, starting from thiophene, and characterized. Polymers containing these electroative side groups were then prepared by two strategies. The oligomers were attached to existing polymer systems and were connected to a polymerizable unit leading to monomer containing the oligothiophenes as side groups. Subsequently the properties of the monomers and the polymers were investigated. A butylcellulose derivative carrying terthienyl side chains (BCTTE, 26) was synthesized starting from cellulose acetate and 5-(2-chloroethyl)-2,2':5',2'-terthiophene (4). The polymer had a degree of substitution (DS) of the butyl and terthienyl side chains of DSbutyl = 1.9 and DSterth = 0.35, respectively. It was successfully spread on a Langmuir-Blodgett (LB) trough and then transferred to several solid substrates. X-rays reflectometry showed an ordered architecture of the cellulose backbones. However, the terthiophene side groups were found as isotropically aligned by polarized UV-Vis spectroscopy. When used as anode material in the electropolymerization of 3-pentylthiophene (28), polythiophene was grafted onto the cellulose backbone through the terthienyl side groups. The polythiophene chains showed an average anisotropic alignment of 20 % along the LB dipping direction, calculated by means of polarized UV-Vis spectroscopy. A second butylcellulose derivative carrying sexithienyl side chains (BCST) was synthesized and investigated, starting from butylcellulose and 2-[b ', b ''-dipentyl-5'''-(2-hydroxyethyl)-2,2': 5',2':5',2'':5'',2'':5'',2'''-sexithiophen-5-yl]-ethyl p-toluensulfonate (7). The polymer showed formation of stable LB monolayers at the air-water interface, but its transfer onto solid substrates was not successful. A poly(p-phenylene-ethynylene) bearing sexithienyl side chains (BzAcST, 31) was prepared by reaction of the two monomers 2-[b ', b ''-dipentyl-5'''-(2-hydroxyethyl)-2,2': 5',2':5',2'':5'',2'':5'',2'''-sexithiophen-5-yl]-ethyl 2,5-diiodobenzoate (15) and 2-[b', b ''-dipentyl-5'''-(2-hydroxyethyl)-2,2':5',2':5',2'':5'',2'':5'',2'''-sexithiophen-5-yl]-ethyl 2,5-diethynylbenzoate (18). The polymer was obtained as insoluble product. Upon oxidation with FeCl3 (doping) of the polymer suspension, BzAcST showed an electrical conductivity of ó = 2.5 . 10 -6 S/cm, a typical value for semiconductors. The IR spectrum of the doped polymer presented the diagnostic bands of oxidized sexithiophene in good agreement with literature results. Along with the monomer and polymer synthesis, an a,a '-disubstituted sexithiophene, b ', b ''-dipentyl-5,5'''-bis-(2-hydroxyethyl)-2,2':5',2':5',2'':5'',2'':5'',2'''-sexithiophene (6a),was synthesized and characterized. The UV-Vis absorption of the chromophore wasinvestigated as a function of temperature and different solvents, showing a blue-shift of the absorption maximum with increasing temperature and a red-shift changing the solvent from hexane to ethanol to toluene. Monitoring the change of the UV-Vis spectrum upon electrochemical oxidation, the oxidized chromophore showed a new broad absorption band, red shifted with respect to the p -p* transition of the neutral state. Upon reduction, the new band disappeared and the UV-Vis spectrum of the chromophore was restored. Such oxidation-reduction cycles were totally reversible. This feature, together with the absorption maximum falling in the visible region, makes this chromophore a suitable compound for the development of an electrochemical sensor.Attempts to polymerize acrylic monomers carrying sexythienyl side chains both via radical polymerization, as in the case of 2-[b ', b ''-dipentyl-5'''-(2-hydroxyethyl)-2,2': 5',2':5',2'':5'',2'':5'',2'''-sexithiophen-5-yl]-ethyl acrylate (8), and anionic polymerization, as in the case of 2-{b ', b ''-dipentyl-5'''-[2-(tertbutyldimethylsiloxy)ethyl]-2,2':5',2':5',2'':5'',2'': 5'',2''' -sexithiophen-5-yl}-ethylacrylate (29), were not successful, probably due to the steric hindrance of the oligothiophene side group. However, due to the time consuming and therefore restricted availability of the monomers, a screening of the polymerization conditions towards the formation of polymeric material was not possible.

Kinetics study on melt grafting copolymerization of LLDPE with acid monomers using reactive extrusion method

Graft copolymerization in the molten state is of fundamental importance as a probe of chemical modification and reactive compatibilization. However, few grafting kinetics studies on reactive extrusion were carried out for the difficulties as expected. In this work, the macromolecular peroxide-induced grafting of acrylic acid and methyl methacrylate onto linear low density polyethylene by reactive extrusion was chosen as the model system for the kinetics study; the samples were taken out from the barrel at five ports along screw axis and analyzed by FTIR, H-1 NMR, and ESR. For the first time, the time-evolution of reaction rate, the reaction order, and the activation energy of graft copolymerization and homopolymerization in the twin screw extruder were directly obtained. On the basis of these results, the general reaction mechanism was tentatively proposed. It was demonstrated that an amount of chain propagation free radicals could keep alive for several minutes even the peroxides completely decomposed and the addition of monomer to polymeric radicals was the rate-controlled step for the graft copolymerization.

Morphology, structure, and rheological property of linear low-density polyethylene grafted with acrylic acid

The chain structure, spherulite morphology, and theological property of LL-DPE-g-AA were studied by using electronspray mass spectroscopy, C-13-NMR, and rheometer. Experimental evidence proved that AA monomers grafted onto the LLDPE backbone formed multiunit AA branch chains. It was found that AA branch chains could hinder movement of the LLDPE main chain during crystallization. Spherulites of LLDPE became more anomalous because of the presence of AA branch chains. Rheological behavior showed that AA branch chains could act as an inner plasticizer at the temperature range of 170-200 degreesC, which made LLDPE-g-AA easy to further process. (C) 2001 John Wiley & Sons, Inc.

WATER-SOLUBLE AND AMPHIPHILIC POLYMERS .11. GRAFT-POLYMERIZATION OF ACRYLIC-ACID ON POLY(VINYL ALCOHOL)

The graft polymerization of acrylic acid(AA) on poly(vinyl alcohol) (PVAL) has been investigated by using either potassium persulfate (KPS) or ceric ammonium nitrate(CAN) as an initiator. In the case of KPS initiation, the formation of the graft polymer always lags behind the homopolymer formation. The graft polymer is separated by acetone, and the increase of reaction temperature favors the homopolymer formation at the early stage. In the case of CAN initiation, graft polymers with a high PAA content can hardly be obtained when the polymerization is performed under nitrogen and at < 0.06 mol/L HNO3 concentration. It has been found that incorporation of a small amount of oxygen in a protective nitrogen gas accelerates markedly the graft polymerization, and that the resulting graft polymers can not be separated by acetone precipitation technique in most cases. The Dalian nitrogen(containing 0.7% oxygen) is a good protective gas for CAN-initiated PVAL-AA graft polymerization.

Development and application of methods for determination of residual monomer in dental acrylic resins using high performance liquid chromatography

Two high-performance liquid chromatographic methods for determination of residual monomer in dental acrylic resins are described. Monomers were detected by their UV absorbance at 230 nm, on a Nucleosil((R)) C-18 (5 mu m particle size, 100 angstrom pore size, 15 x 0.46 cm i.d.) column. The separation was performed using acetonitrile-water (55:45 v/v) containing 0.01% triethylamine (TEA) for methyl methacrylate and butyl methacrylate, and acetonitrile-water (60:40 v/v) containing 0.01% TEA for isobutyl methacrylate and 1,6-hexanediol dimethacrylate as mobile phases, at a flow rate of 0.8 mL/min. Good linear relationships were obtained in the concentration range 5.0-80.0 mu g/mL for methyl methacrylate, 10.0-160.0 mu g/mL for butyl methacrylate, 50.0-500.0 mu g/mL for isobutyl methacrylate and 2.5-180.0 mu g/mL for 1,6-hexanediol dimethacrylate. Adequate assay for intra- and inter-day precision and accuracy was observed during the validation process. An extraction procedure to remove residual monomer from the acrylic resins was also established. Residual monomer was obtained from broken specimens of acrylic disks using methanol as extraction solvent for 2 h in an ice-bath. The developed methods and the extraction procedure were applied to dental acrylic resins, tested with or without post-polymerization treatments, and proved to be accurate and precise for the determination of residual monomer content of the materials evaluated. Copyright (c) 2005 John Wiley & Sons, Ltd.

Cytotoxicity of monomers, plasticizer and degradation by-products released from dental hard chairside reline resins

Objectives. The aim of this study was to evaluate the cytotoxic effect of the monomers isobutyl methacrylate (IBMA) and 1,6-hexanediol dimethacrylate (1,6-HDMA), the plasticizer di-n-butyl phthalate (DBP), and the degradation by-products methacrylic acid (MA) and benzoic acid (BA) on L929 cells. Based on previous investigations on the release of these compounds from hard chairside reline resins, a range of concentrations (mu mol/L) were selected for the cytotoxicity tests (IBMA, 5.491406.57; 1,6-HDMA, 1.2239.32; DBP, 1.12143.8; MA, 9.07581; BA, 3.19409).Methods. Cytotoxic effects were assessed using MTT and 3H-thymidine assays after the cells had been exposed to the test compounds at the given concentrations for 24h. Cytotoxicity was rated based on cell viability relative to controls (cells exposed to medium without test substances).Results. DNA synthesis activity was inhibited by all compounds. Mitochondrial dehydrogenase activity decreased in cells treated with monomers, plasticizer and MA by-product, whereas no cytotoxic effect was observed on contact with BA at the majority of concentrations tested. The ranges of suppression for 3H-thymidine assay were: IBMA, 2595%; 1,6-HDMA, 9598%; DBP, 4098%; MA, 9799%; BA, 5471%. For MTT assay, the ranges of suppression were: IBMA, 096%; 1,6-HDMA, 2689%; DBP, 1780%; MA, 5266%; BA, 027%. The 3H-thymidine assay was more sensitive than the MTT assay.Significance. This study evaluated the cytotoxicity of a wide range of concentrations of monomers (IBMA and 1,6-HDMA), plasticizer (DBP) and degradation by-products (MA and BA), including those expected to be released from hard chairside reline resins. The differences observed in the cytotoxicity of these compounds, along with other properties, may assist the dental practitioners in the selection of reline materials with improved service life performance and low risk of adverse reactions in patients who wear relined dentures.

Candida albicans adherence to an acrylic resin modified by experimental photopolymerised coatings: An in vitro study

Purpose: This study evaluated whether photopolymerised coatings containing zwitterion or hydrophilic monomers would reduce the adhesion of Candida albicans to an acrylic resin. Materials and methods: Disc-shaped samples (n = 468) were fabricated with rough or smooth surfaces. The samples did not receive any surface treatment (control) or were coated with one of the following experimental coatings (2-hydroxyethyl methacrylate - HE; 3-hydroxypropyl methacrylate - HP; and 2-trimethylammonium ethyl methacrylate chloride - T; and sulfobetaine methacrylate - S). The concentrations of the constituent monomers were 25, 30 or 35%. The water contact angles of the samples were measured, and half of the samples were exposed to saliva. The adherent yeast cells were counted after crystal violet staining. Results: For the smooth samples, the groups S35, HP35 and HE35 showed significantly lower number of adhered Candida than control, in the absence of saliva. There were no significant differences among the experimental and control groups for the rough samples, but the saliva decreased the cell numbers for groups S25, S30 and HP30. The photoelectron spectroscopy analysis confirmed the changes in the chemical compositions of the experimental samples. Conclusions: The experimental photopolymerised coatings changed the chemical composition and decreased C. albicans adhesion in the groups S35, HP35 and HE35, suggesting that they should be further investigated. © 2012 The Gerodontology Society and John Wiley & Sons A/S.

Effect of experimental photopolymerized coatings on the hydrophobicity of a denture base acrylic resin and on Candida albicans adhesion

Objective: This study investigated the effect of experimental photopolymerized coatings, containing zwitterionic or hydrophilic monomers, on the hydrophobicity of a denture base acrylic resin and on Candida albicans adhesion. Methods: Acrylic specimens were prepared with rough and smooth surfaces and were either left untreated (control) or coated with one of the following experimental coatings: 2-hydroxyethyl methacrylate (HE); 3-hydroxypropyl methacrylate (HP); and 2-trimethylammonium ethyl methacrylate chloride (T); and sulfobetaine methacrylate (S). The concentrations of these constituent monomers were 25%, 30% or 35%. Half of the specimens in each group (control and experimentals) were coated with saliva and the other half remained uncoated. The surface free energy of all specimens was measured, regardless of the experimental condition. C. albicans adhesion was evaluated for all specimens, both saliva conditioned and unconditioned. The adhesion test was performed by incubating specimens in C. albicans suspensions (1 × 10 7 cell/mL) at 37 °C for 90 min. The number of adhered yeasts were evaluated by XTT (2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-5-[{phenylamino} carbonyl]-2H-tetrazolium-hydroxide) method. Results: For rough surfaces, coatings S (30 or 35%) and HP (30%) resulted in lower absorbance values compared to control. These coatings exhibited more hydrophilic surfaces than the control group. Roughness increased the adhesion only in the control group, and saliva did not influence the adhesion. The photoelectron spectroscopy analysis (XPS) confirmed the chemical changes of the experimental specimens, particularly for HP and S coatings. Conclusions: S and HP coatings reduced significantly the adhesion of C. albicans to the acrylic resin and could be considered as a potential preventive treatment for denture stomatitis. © 2012 Elsevier Ltd.

In vitro evaluation of adherence of Candida albicans, Candida glabrata, and Streptococcus mutans to an acrylic resin modified by experimental coatings

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Acrylic resin cytotoxicity for denture base - Literature review

Acrylic resin is a widely used material in clinical practice, and a satisfactory biocompatibility is essential. When the resin polymerization reaction is incomplete, residual monomers are released into the oral cavity. The aim of this study was to evaluate, through a literature review, the cytotoxicity caused by the denture base acrylic resin used, and its components. The selection of published studies was performed on the Pubmed database from January 2008 to July 2013. The keywords used were: cytotoxicity and acrylic resins, cytotoxicity and denture base resins and cytotoxicity and oral prosthesis. Inclusion criteria were: in vitro studies and literature reviews published in English that evaluated the acrylic resin cytotoxicity for denture base and its components. Studies with no reference to the search strategy were excluded. A total of 182 articles were found. Among these, only 13 were included for writing this review. The MTT test is the most common test used to evaluate acrylic resin cytotoxicity. Auto-polymerized resin is more cytotoxic than heat-polymerized resin because of its higher quantity of residual monomers which cause cell and tissue changes in the oral mucosa. However, more studies are necessary for the development of biocompatible materials.

Effect of poly(acrylic acid) end-group functionality on inhibition of calcium oxalate crystal growth

A number of series of poly(acrylic acids) (PAA) of differing end-groups and molecular weights prepared using atom transfer radical polymerization were used as inhibitors for the crystallization of calcium oxalate at 23 and 80°C. As measured by turbidimetry and conductivity and as expected from previous reports, all PAA series were most effective for inhibition of crystallization at molecular weights of 1500–4000. However, the extent of inhibition was in general strongly dependent on the hydrophobicity and molecular weight of the end-group. These results may be explicable in terms of adsorption/desorption of PAA to growth sites on crystallites. The overall effectiveness of the series didn't follow a simple trend with end-group hydrophobicity, suggesting self-assembly behavior or a balance between adsorption and desorption rates to crystallite surfaces may be critical in the mechanism of inhibition of calcium oxalate crystallization.