A novel electrochemical biosensing method with double-layered polymer brush modified electrode.

We developed a novel electrochemical biosensor electrode that has a potential to reduce background noise for which we constructed an original conductive substrate modified with a double-layered polymer brush structure that is water impermeable and can control biomolecules adsorption/desorption. In this study, a hydrophobic poly(tert-butyl methacrylate) brush layer was prepared on a gold electrode, and then, the tert-butyl group near the outermost surface was dissociated by the acid treatment to obtain a hydrophilic carboxy group, thereby fabricating a conductive substrate with the double-layered polymer brush structure. Formation of the double-layered polymer brush structure was indicated by surface wettability and optical analyses. The potential difference and hydrogen ion concentration, which is a typical parameter of the surrounding environment, were linearly correlated with the gold electrode having a double-layered polymer brush structure with carboxyl groups. However, there was no correlation on gold electrodes with self-assembled monolayers presenting carboxy groups. It is considered that the pH responsiveness of the carboxy groups on the outermost surface could be exhibited remarkably because the charge state in the vicinity of the surface became constant due to the hydrophobic polymer brush layer having a certain thickness. The target DNA could be captured more efficiently at the probe DNA-immobilized electrode with the double-layered polymer brush structure than when using COOH-SAM. This is the first report of the application of the double-layered polymer brush structure for the electrochemical biosensing, and it will be an excellent surface modification method to reduce background noise.

Verification of the effect of acquisition time for SwiftScan on quantitative bone single-photon emission computed tomography using an anthropomorphic phantom.

BACKGROUND: SwiftScan single-photon emission computed tomography (SPECT) is a recently released scanning technique with data acquired when the detector is stationary and when it moves from one view to the next. The influence of scan time for using SwiftScan on quantitative bone SPECT remains unclear. This study aimed to clarify the effect of the scan time for SwiftScan SPECT on the image quality and quantification of bone SPECT compared to step and shoot mode (SSM) using 99mTc-filled anthropomorphic phantom (SIM2 bone phantom). MATERIALS AND METHODS: Phantom SPECT/computed tomography (CT) images were acquired using Discovery NM/CT 860 (GE Healthcare) with a low-energy high-resolution sensitivity collimator. We used the fixed parameters (subsets 10 and iterations 5) for reconstruction. The coefficient of variation (CV), contrast-to-noise ratio (CNR), full width at half maximum (FWHM), and quantitative value of SwiftScan SPECT and SSM were compared at various acquisition times (5, 7, 17, and 32 min). RESULTS: In the short-time scan (< 7 min), the CV and CNR of SwiftScan SPECT were better than those of SSM, whereas in the longtime scan (> 17 min), the CV and CNR of SwiftScan SPECT were similar to those of SSM. The FWHMs for SwiftScan SPECT (13.6-14.8 mm) and SSM (13.5-14.4 mm) were similar. The mean absolute errors of quantitative values at 5, 7, 17, and 32 min were 38.8, 38.4, 48.8, and 48.1, respectively, for SwiftScan SPECT and 41.8, 40.8%, 47.2, and 49.8, respectively, for SSM. CONCLUSIONS: SwiftScan on quantitative bone SPECT provides improved image quality in the short-time scan with quantification similar to or better than SSM. Therefore, in clinical settings, using SwiftScan SPECT instead of the SSM scan protocol in the short-time scan might provide higher-quality diagnostic images than SSM. Our results could provide vital information on the use of SwiftScan SPECT.

Effect of liposome surface modification with water-soluble phospholipid polymer chain-conjugated lipids on interaction with human plasma proteins.

Alternative liposome surface coatings for PEGylation to evade the immune system, particularly the complement system, have garnered significant interest. We previously reported poly(2-methacryloyloxyethyl phosphorylcholine) (MPC)-based lipids (PMPC-lipids) and investigated the surface modification of liposomes. In this study, we synthesize PMPC-lipids with polymerization degrees of 10 (MPC10-lipid), 20 (MPC20-lipid), 50 (MPC50-lipid), and 100 (MPC100-lipid), and coated liposomes with 1, 5, or 10 mol% PMPC-lipids (PMPC-liposomes). Non-modified and PEGylated liposomes are used as controls. We investigate the liposome size, surface charge, polydispersity index, and adsorption of plasma proteins to the liposomes post incubation in human plasma containing N,N,N',N'-ethylenediamine tetraacetic acid (EDTA) or lepirudin by some methods such as sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), western blotting, and automated capillary western blot, with emphasis on the binding of complement protein C3. It is shown that the coating of liposome PMPC-lipids can suppress protein adsorption more effectively with an increase in the molecular weight and molar ratio (1-10 mol%). Apolipoprotein A-I is detected on PMPC-liposomes with a higher molecular weight and higher molar ratio of PMPC-lipids, whereas α2-macroglobulin is detected on non-modified, PEGylated, and PMPC-liposomes with a shorter polymer chain. In addition, a correlation is shown among the PMPC molecular weight, molar ratio, and C3 binding. The MPC10-lipid cannot inhibit C3 binding efficiently, whereas surface modifications with 10 mol% MPC20-lipid and 5 mol% and 10 mol% MPC50-lipid suppress both total protein and C3 binding. Hence, liposome modification with PMPC-lipids can be a possible strategy for avoiding complement activation.

In Situ Electrical Monitoring of Methylated DNA Based on Its Conformational Change to G-Quadruplex Using a Solution-Gated Field-Effect Transistor.

Methylated DNA is not only a diagnostic but also a prognostic biomarker for early-stage cancer. However, sodium bisulfite sequencing as a "gold standard" method for detection of methylation markers has some drawbacks such as its time-consuming and labor-intensive procedures. Therefore, simple and reliable methods are required to analyze DNA sequences with or without methylated residues. Herein, we propose a simple and direct method for detecting DNA methylation through its conformation transition to G-quadruplex using a solution-gated field-effect transistor (SG-FET) without using labeled materials. The BCL-2 gene, which is involved in the development of various human tumors, contains G-rich segments and undergoes a conformational change to G-quadruplex depending on the K+ concentration. Stacked G-quadruplex strands move close to the SG-FET sensor surface, resulting in large electrical signals based on intrinsic molecular charges. In addition, a dense hydrophilic polymer brush is grafted using surface-initiated atom transfer radical polymerization onto the SG-FET sensor surface to reduce electrical noise based on nonspecific adsorption of interfering species. In particular, control of the polymer brush thickness induces electrical signals based on DNA molecular charges in the diffusion layer, according to the Debye length limit. A platform based on the SG-FET sensor with a well-defined polymer brush is suitable for in situ monitoring of methylated DNA and realizes a point-of-care device with a high signal-to-noise ratio and without the requirement for additional processes such as bisulfite conversion and polymerase chain reaction.

Detection of CpG Methylation in G-Quadruplex Forming Sequences Using G-Quadruplex Ligands.

Genomic DNA methylation is involved in many diseases and is expected to be a specific biomarker for even the pre-symptomatic diagnosis of many diseases. Thus, a rapid and inexpensive detection method is required for disease diagnosis. We have previously reported that cytosine methylation in G-quadruplex (G4)-forming oligonucleotides develops different G4 topologies. In this study, we developed a method for detecting CpG methylation in G4-forming oligonucleotides based on the structural differences between methylated and unmethylated G4 DNAs. The differences in G4 topologies due to CpG methylation can be discriminated by G4 ligands. We performed a binding assay between methylated or unmethylated G4 DNAs and G4 ligands. The binding abilities of fluorescent G4 ligands to BCL-2, HRAS1, HRAS2, VEGF G4-forming sequences were examined by fluorescence-based microtiter plate assay. The differences in fluorescence intensities between methylated and unmethylated G4 DNAs were statistically significant. In addition to fluorescence detection, the binding of G4 ligand to DNA was detected by chemiluminescence. A significant difference was also detected in chemiluminescence intensity between methylated and unmethylated DNA. This is the first study on the detection of CpG methylation in G4 structures, focusing on structural changes using G4 ligands.

Synthesis of poly(2-methacryloyloxyethyl phosphorylcholine)-conjugated lipids and their characterization and surface properties of modified liposomes for protein interactions.

Poly(ethylene glycol) (PEG) is frequently used for liposomal surface modification. However, as PEGylated liposomes are cleared rapidly from circulation upon repeated injections, substitutes of PEG are being sought. We focused on a water-soluble polymer composed of 2-methacryloyloxyethyl phosphorylcholine (MPC) units, and synthesized poly(MPC) (PMPC)-conjugated lipid (PMPC-lipid) with degrees of MPC polymerization ranging from 10 to 100 (calculated molecular weight: 3 to 30 kDa). In addition, lipids with three different alkyl chains, myristoyl, palmitoyl, and stearoyl, were applied for liposomal surface coating. We studied the interactions of PMPC-lipids with plasma albumin, human complement protein C3 and fibrinogen using a quartz crystal microbalance with energy dissipation, and found that adsorption of albumin, C3 and fibrinogen could be suppressed by coating with PMPC-lipids. In particular, the effect was more pronounced for PMPC chains with higher molecular weight. We evaluated the size, polydispersity index, surface charge, and membrane fluidity of the PMPC-lipid-modified liposomes. We found that the effect of the coating on the dispersion stability was maintained over a long period (98 days). Furthermore, we also demonstrated that the anti-PEG antibody did not interact with PMPC-lipids. Thus, our findings suggest that PMPC-lipids can be used for liposomal coating.

Direct photoreactive immobilization of water-soluble phospholipid polymers on substrates in an aqueous environment.

The purpose of this study is to achieve a simpler and safer surface modification of substrates using a photoreactive polymer in an aqueous environment. We synthesized water-soluble photoreactive polymers with both phenylazide groups and phosphorylcholine groups, poly(2-methacryloyloxyethyl phosphorylcholine-co-4-methacryl tetra(ethylene glycol)oxycarbonyl-4-phenylazide) (PMEPAz), via reversible addition fragmentation chain transfer polymerization. PMEPAz with different polymerization degrees were synthesized with a well-defined structure. To immobilize PMEPAz on the substrate surface by photoreaction, it is necessary to adsorb the polymer on the substrate surface in an aqueous solution because the phenylazide groups chemically bind to the substrate via a hydrogen abstract reaction. The relationship between the polymer solubilization state in the aqueous solution and the adsorption behavior at the surface was investigated. PMEPAz began to form unstable molecular aggregates at a concentration of 10-2 mg/mL and formed stable aggregates at 100 mg/mL. At a concentration of 10-1 mg/mL, unstable molecular aggregates of PMEPAz were formed in the aqueous solution, resulting in the maximization of the amount of adsorbed polymer and effective photoreaction with the substrate. The thickness of the reacted polymer layer on the substrate increased with an increase in the polymerization degree, a uniform polymer layer with a thickness of 3.4 nm was formed when the polymerization degree was 400. After surface modification, the hydrophobic surfaces of the original substrates became hydrophilic. Additionally, fibrinogen adsorption and platelet adhesion were effectively suppressed based on the characteristics of the phosphorylcholine unit.

Effects of molecular architecture of photoreactive phospholipid polymer on adsorption and reaction on substrate surface under aqueous condition.

Water-soluble photoreactive polymers with both phosphorylcholine and benzophenone groups were synthesized for the reaction between the polymers and the substrate in aqueous medium. To control the polymer architecture, the living radical polymerization method was applied to the copolymerization of 2-methacryloyloxyethyl phosphorylcholine and benzophenone methacrylates. These polymers possess various architectures, such as linear polymers, polymers with hydrophobic terminals, and 4-armed star-like polymers, that could promote their adsorption on the substrate surfaces. Additionally, two types of benzophenone groups were examined. Due to the bulky phosphorylcholine group, tetra(ethylene oxide) group as a spacer between polymer main chain and benzophenone group was considered. These polymers could adsorb on the surface in an aqueous medium, followed by reaction on the surface via photoirradiation depending on the chemical structure of the benzophenone group. The thickness of the polymer layer depended on the polymer architecture, i.e. a polymer with a hydrophobic terminal could form a thick layer. After modification, the contact angle by air in the aqueous medium decreased, compared to that on the base substrate. This was due to the hydrophilic nature based on the phosphorylcholine groups at the surface. The amount of proteins adsorbed on the surface also decreased because of the surface modification. These findings indicated that these water-soluble photoreactive polymers could be applied for the safer and effective surface modification of substrates via conventional photoirradiation without using an organic solvent.

Water-Soluble and Cytocompatible Phospholipid Polymers for Molecular Complexation to Enhance Biomolecule Transportation to Cells in Vitro.

Water-soluble and cytocompatible polymers were investigated to enhance a transporting efficiency of biomolecules into cells in vitro. The polymers composed of a 2-methacryloyloxyethyl phosphorylcholine (MPC) unit, a hydrophobic monomer unit, and a cationic monomer unit bearing an amino group were synthesized for complexation with model biomolecules, siRNA. The cationic MPC polymer was shown to interact with both siRNA and the cell membrane and was successively transported siRNA into cells. When introducing 20-50 mol% hydrophobic units into the cationic MPC polymer, transport of siRNA into cells. The MPC units (10-20 mol%) in the cationic MPC polymer were able to impart cytocompatibility, while maintaining interaction with siRNA and the cell membrane. The level of gene suppression of the siRNA/MPC polymer complex was evaluated in vitro and it was as the same level as that of a conventional siRNA transfection reagent, whereas its cytotoxicity was significantly lower. We concluded that these cytocompatible MPC polymers may be promising complexation reagent for introducing biomolecules into cells, with the potential to contribute to future fields of biotechnology, such as in vitro evaluation of gene functionality, and the production of engineered cells with biological functions.

Effects of molecular interactions at various polymer brush surfaces on fibronectin adsorption induced cell adhesion.

The effects of protein adsorption on the polymer brush surfaces with well-defined chemical structures and physical properties were examined with respect to initial protein adsorption, structural changes to the adsorbed proteins, and subsequent cell adhesion. Four polymer brush surfaces with different hydrophilicities and charge states were prepared. The molecular interaction forces during adsorption-desorption processes of protein on the polymer brush surfaces depending on the chemical structure of the polymer were determined. Crucially, these molecular interactions affected the adsorption behavior and structural changes of fibronectin (FN), a cell-adhesive protein, used in this study. Adsorption of FN onto the zwitterionic polymer and anionic polymer surfaces was difficult, however significant protein adsorption to the hydrophobic and cationic surfaces was observed. Further, the structural changes to the adhered FN on these surfaces were significant. Subsequent cell adhesion experiments revealed that the adhered cell density was correlated with the amount of adsorbed FN and the degree of FN structural change. In addition, the cationic surface inhibited cell proliferation behavior. These results indicate that cellular responses can be indirectly regulated by controlling the molecular interactions which induced the structural change of adsorbed proteins via the material surface properties.

Interface of Phospholipid Polymer Grafting Layers to Analyze Functions of Immobilized Oligopeptides Involved in Cell Adhesion.

The aim of this study was to design a material surface for use in the analysis of the behavior of biomolecules at the interface of direct cell contact. A superhydrophilic surface was prepared with poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC), which was grafted onto a substrate with controlled polymer chain density. An arginine-glycine-aspartic acid (RGD) peptide was immobilized at the surface of the polymer graft surface (PMPC-RGD surface). Initial adhesion of the cells to this substrate was observed. The PMPC-RGD surface could enable cell adhesion only through RGD peptide-integrin interactions. The density and movability of the RGD peptide at the terminal of the graft PMPC chain and the orientation of the RGD peptide affected the density of adherent cells. Thus, the PMPC graft surface may be a good candidate for a new platform with the ability to immobilize biomolecules to a defined position and enable accurate analysis of their effects on cells.

Hybridization of a phospholipid polymer hydrogel with a natural extracellular matrix using active cell immobilization.

Three-dimensional tissue organization is still an obstacle in the field of tissue engineering, which generally involves cell immobilization, proliferation, and organization. As an artificial extracellular matrix (ECM) for providing a suitable environment of cells to construct tissues, combination of cytocompatible polymer hydrogels and natural ECM produced by the immobilized cells was considered. In this research, we designed a spontaneously forming hydrogel system between two water-soluble polymers for the immobilization of cells. These polymers were poly(2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate-co-p-vinylphenylboronic acid-co-N-succinimidyloxycarbonyl tetra(ethylene glycol)methacrylate) (PMBVS) and poly(vinyl alcohol) (PVA) to form a PMBVS/PVA hydrogel in a cell culture medium under mild conditions. Basic fibroblast growth factor (bFGF) was conjugated with PMBVS (PMBV-bFGF). To enhance the growth of the immobilized cells, mouse fibroblast L929 cells were immobilized in the PMBVS/PVA hydrogel and the PMBV-bFGF/PVA hydrogel, and their proliferation and secretion of the ECM under stimulation with bFGF was observed. The ECM infiltrated and replaced the hydrogel, resulting in the formation of a hybrid hydrogel with the ECM and laden cells.

Modification of human MSC surface with oligopeptide-PEG-lipids for selective binding to activated endothelium.

Promising cell therapies using mesenchymal stem cells (MSCs) is proposed for stroke patients. Therefore, we aimed to efficiently accumulate human MSC (hMSC) to damaged brain area to improve the therapeutic effect using poly(ethylene glycol) (PEG)-conjugated phospholipid (PEG-lipid) carrying an oligopeptide as a ligand, specific for E-selectin which is upregulated on activated endothelial cells under hypoxia-like stroke. Here we synthesized E-selectin-binding oligopeptide (ES-bp) conjugated with PEG spacer having different molecular weights from 1 to 40 kDa. We found that ES-bp can be immobilized onto the hMSC surface through PEG-lipid without influence on cell growth and differentiation into adipocytes and osteocytes, respectively. It is also possible to control the immobilization of ES-bp on hMSC surface (<108 ES-bp per cell). Immobilized ES-bp can be continuously immobilized at the outside of cell membrane when PEG-lipids with PEG 5 and 40 kDa were used. In addition, the modified hMSC can specifically attach onto E-selectin-immobilized surface as a model surface of activated endothelium in human blood, indicating the sufficient number of immobilized ES-bp onto hMSC. Thus, this technique is one of the candidates for hMSC accumulation to cerebral infarction area. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1779-1792, 2019.

Validation of an MPC Polymer Coating to Attenuate Surface-Induced Crosstalk between the Complement and Coagulation Systems in Whole Blood in In Vitro and In Vivo Models.

Artificial surfaces that come into contact with blood induce an immediate activation of the cascade systems of the blood, leading to a thrombotic and/or inflammatory response that can eventually cause damage to the biomaterial or the patient, or to both. Heparin coating has been used to improve hemocompatibility, and another approach is 2-methacryloyloxyethyl phosphorylcholine (MPC)-based polymer coatings. Here, the aim is to evaluate the hemocompatibility of MPC polymer coating by studying the interactions with coagulation and complement systems using human blood in vitro model and pig in vivo model. The stability of the coatings is investigated in vitro and MPC polymer-coated catheters are tested in vivo by insertion into the external jugular vein of pigs to monitor the catheters' antithrombotic properties. There is no significant activation of platelets or of the coagulation and complement systems in the MPC polymer-coated one, which was superior in hemocompatibility to non-coated matrix surfaces. The protective effect of the MPC polymer coat does not decline after incubation in human plasma for up to 2 weeks. With MPC polymer-coated catheters, it is possible to easily draw blood from pig for 4 days in contrast to the case for non-coated catheters, in which substantial clotting is seen.

Surface functionalization of polytetrafluoroethylene substrate with hybrid processes comprising plasma treatment and chemical reactions.

Polytetrafluoroethylene (PTFE) exhibits excellent mechanical properties and chemical stability and has been widely used in medical fields for the preparation of implantable medical devices. However, the implantation of PTFE in living systems results in inflammation reactions and infections at the surface thus limits its long-term applications. For PTFE surface modification, we examined the effects of mussel-inspired polydopamine (PDA) coating and the further introduction of functional groups. During PDA coating, the plasma pretreatment on PTFE enhanced the stability of the PDA coating layer. Furthermore, the introduction of functional groups on the PDA layer was carried out using reactive functional groups for the photoinduced graft polymerization of methacrylate. For instance, 2-methacryloyloxyethyl phosphorylcholine (MPC) could be polymerized from the surface of the substrate. These chemical modifications were confirmed step by step using spectroscopes to obtain the hydrophilic surface of the poly(MPC)-modified PTFE. The protein adsorption behaviors on PTFE and poly(MPC)-modified PTFE were compared to understand biocompatibility characteristics of these substrates. The surface of PTFE was immediately covered with albumin and the contact between the substrate and the serum resulted in an increase in the fibrinogen composition with time. On the other hand, fewer proteins were adsorbed on the poly(MPC)-modified PTFE substrate. Thus, this modification procedure would serve as a strategy for safer alterations in PTFE surfaces to expand the life span of the PTFE-carrying medical devices in living systems.

Photoinduced Surface Zwitterionization for Antifouling of Porous Polymer Substrates.

Surface functionalization of polymeric porous substrates is one of the most important requirements to enhance their applications in the biomedical field. In this study, we achieved photoinduced surface modification using a highly efficient reaction of hydrophilic polymers bearing phosphorylcholine groups. Polymers composed of 2-methacryloyloxyethyl phosphorylcholine (MPC) units and 2-( N-ethylanilino)ethyl methacrylate units were synthesized with attention to the polymer architectures. The surface modification of the porous polyethylene (PE) substrates was carried out by the coating of the MPC polymers with a photochemical radical generator, followed by photoirradiation for a few minutes. Surface analysis by attenuated total reflectance Fourier transform IR spectroscopy and X-ray photoelectron spectroscopy indicated that the MPC polymer layer was generated on the PE surface. Cross-sectional confocal microscopy images showed that the MPC polymers were coated on the polymer surface, even inside the porous structure of the PE substrate. After modification, the porous PE substrates showed a significant increase in hydrophilicity and the water-penetration rate through the pores. Furthermore, the amount of protein adsorbed on the PE substrate was reduced significantly by the surface modification. These functionalities were dependent on the MPC polymer architectures. Thus, we concluded that the photoreactive polymer system developed furnished the porous substrates with antifouling properties.

Initial Cell Adhesion onto a Phospholipid Polymer Brush Surface Modified with a Terminal Cell Adhesion Peptide.

Dynamic changes in the properties of adsorbed protein layers at material surfaces make it difficult to analyze a cell adhesion behavior. Adhesion is affected by the ligand molecules in the adsorbed protein layers on the material's surface. This study aimed to quantitatively analyze the initial cell adhesion onto a polymeric surface modified with immobilized cell adhesion molecules with a well-defined structure. Peptides containing an arginine-glycine-aspartic acid (RGD) sequence were introduced at almost all the termini of the grafted poly(2-methacryloyloxyethyl phosphorylcholine) [poly(MPC)] chains using a click reaction at a highly protein-resistant poly(MPC) brush layer. Thus, the surface could bind to the cell membrane proteins only through the immobilized RGD. Furthermore, the degree of polymerization of the grafted poly(MPC) chains could control the hydrated poly(MPC) brush layer softness, as determined by measuring the dissipation energy loss using a quartz crystal microbalance. At the initial stage of cell adhesion, the density of cells adhering to the RGD-immobilized poly(MPC) brush layers did not depend on the poly(MPC) brush layer softness. However, spreading of the adherent cells was inhibited on the RGD-immobilized poly(MPC) brush layers with a higher softness. Hence, the results suggested that the layer softness did not affect the binding number between the RGD and cell membrane protein during initial cell adhesion; however, the intracellular signaling triggered by the RGD-receptor interaction was inhibited. The poly(MPC) brush surface carrying immobilized cell adhesion molecules has the potential to analyze precisely the effect of the properties of cell adhesion molecules on initial cell adhesion.

Inhibition of denture plaque deposition on complete dentures by 2-methacryloyloxyethyl phosphorylcholine polymer coating: A clinical study.

STATEMENT OF PROBLEM: Denture plaque-associated infections are regarded as a source of serious dental and medical complications in the elderly population. Methods of managing this problem are needed. PURPOSE: The purpose of this clinical study was to evaluate the effects of treatment with a 2-methacryloyloxyethyl phosphorylcholine polymer, PMBPAz, on plaque deposition in complete dentures. MATERIAL AND METHODS: The study protocol was approved by the Ethics Committee of Showa University (#2013-013). Eleven individuals with maxillary complete dentures participated in this study. Their dentures were treated with PMBPAz, and the amount of denture plaque accumulation was evaluated by staining the denture surfaces with methylene blue after 2 weeks of denture usage. The same procedures were repeated to evaluate the original denture surfaces as a control. The image of the stained denture surface was captured using a digital camera, and the percentage of stained area, quantified as a pixel-based density, of the whole denture area (percentage of plaque index) was calculated for the mucosal and polished surfaces. To quantify the biofilm on the dentures, denture plaque biofilm was detached by ultrasonic vibration, resuspended in diluent, and measured with a microplate reader at an optical density of 620 nm. The effects of PMBPAz treatment on these variables were statistically analyzed with ANOVA (α=.05). RESULTS: The mean ±SD percentage of plaque index was 40.7% ±19.9% on the mucosal surfaces and 28.0% ±16.8% on the polished surfaces of the control denture. The mean percentage of plaque index of PMBPAz-treated dentures significantly decreased to 17.4%% ±12.0% on the mucosal surfaces (P<.001) and 15.0% ±9.9% on the polished surfaces (P<.05). The quantification of plaque deposition agreed with the results of these image analyses. CONCLUSIONS: These results demonstrated the effectiveness of the treatment with the PMBPAz to inhibit the bacterial plaque deposition on complete dentures.

A Polymethyl Methacrylate-Based Acrylic Dental Resin Surface Bound with a Photoreactive Polymer Inhibits Accumulation of Bacterial Plaque.

PURPOSE: The aim of this study was to examine the ability of a poly(2-methacryloyloxyethyl phosphorylcholine-co-n-butylmethacrylate-co-2-methacryloyloxyethyloxy-p-azidobenzoate) (PMBPAz) coating on polymethyl methacrylate (PMMA)-based dental resin to inhibit bacterial plaque formation, as well as the polymer's durability against water soaking and chemical exposure. MATERIALS AND METHODS: Successful application of PMBPAz on PMMA surfaces was confirmed by x-ray photoelectron spectroscopy (XPS) and measuring the static air contact angle in water. The anti-adhesive effects to bacterial plaque were evaluated using Streptococcus mutans biofilm formation assay. The mechanical and chemical durabilities of the PMBPAz coating on the PMMA surfaces were examined using soaking and immersion tests, respectively. RESULTS: XPS signals for phosphorus and nitrogen atoms and hydrophilic status on PMMA surfaces treated with PMBPAz were observed, indicating the presence of the polymer on the substrates. The treated PMMA surfaces showed significant inhibition of S mutans biofilm formation compared to untreated surfaces. The PMBPAz coating was preserved after water soaking and chemical exposure. In addition, water soaking did not decrease the ability of treated PMMA to inhibit biofilm formation compared to treated PMMA specimens not subjected to water soaking. CONCLUSION: This study suggests that PMBPAz coating may represent a useful modification to PMMA surfaces for inhibiting denture plaque accumulation.

Solubilization of poorly water-soluble compounds using amphiphilic phospholipid polymers with different molecular architectures.

To achieve stable and effective solubilization of poorly water-soluble bioactive compounds, water-soluble and amphiphilic polymers composed of hydrophilic 2-methacryloyloxyethyl phosphorylcholine (MPC) units and hydrophobic n-butyl methacrylate (BMA) units were prepared. MPC polymers having different molecular architectures, such as random-type monomer unit sequences and block-type sequences, formed polymer aggregates when they were dissolved in aqueous media. The structure of the random-type polymer aggregate was loose and flexible. On the other hand, the block-type polymer formed polymeric micelles, which were composed of very stable hydrophobic poly(BMA) cores and hydrophilic poly(MPC) shells. The solubilization of a poorly water-soluble bioactive compound, paclitaxel (PTX), in the polymer aggregates was observed, however, solubilizing efficiency and stability were strongly depended on the polymer architecture; in other words, PTX stayed in the poly(BMA) core of the polymer micelle formed by the block-type polymer even when plasma protein was present in the aqueous medium. On the other hand, when the random-type polymer was used, PTX was transferred from the polymer aggregate to the protein. We conclude that water-soluble and amphiphilic MPC polymers are good candidates as solubilizers for poorly water-soluble bioactive compounds.