Effects of polymethylmethacrylate nanoplastics on the polychaete Hediste diversicolor: Behavioural, regenerative, and biochemical responses.

Plastics, particularly microplastics (MPs) and nanoplastics (NPs), have been regarded as pollutants of emerging concern due to their effects on organisms and ecosystems, especially considering marine environments. However, in terms of NPs, there is still a knowledge gap regarding the effects of size and polymer on marine invertebrates, such as benthic organisms. Therefore, this study aimed to understand, regarding behavioural, physiological, and biochemical endpoints (neurotransmission, energy metabolism, antioxidant status, and oxidative damage), the effects of 50 nm waterborne polymethylmethacrylate (PMMA) NPs (0.5 to 500 µg/L) on the marine benthic polychaete Hediste diversicolor, a key species in estuarine and coastal ecosystems. Results demonstrated that worms exposed to PMMA NPs had a shorter burrowing time than control organisms. Nevertheless, worms exposed to PMMA NPs (0.5 and 500 µg/L) decreased cholinesterase activity. Energy metabolism was decreased at 50 and 500 µg/L, and glycogen content decreased at all concentrations of PMMA NPs. Enzymes related to the antioxidant defence system (superoxide dismutase and glutathione peroxidase) displayed increased activities in H. diversicolor specimens exposed to concentrations between 0.5 and 500 µg/L, which led to no damage at the cell membrane and protein levels. In this study, polychaetes also displayed a lower regenerative capacity when exposed to PMMA NPs. Overall, the data obtained in this study emphasize the potential consequences of PMMA NPs to benthic worms, particularly between 0.5 and 50 µg/L, with polychaetes exposed to 50 µg/L being the most impacted by the analysed NPs. However, since sediments are considered to be sinks and sources of plastics, further studies are needed to better understand the impacts of different sizes and polymers on marine organisms, particularly benthic species.

UV-B radiation alleviated detrimental effects of polymethyl methacrylate microplastics on marine diatom Thalassiosira pseudonana.

Microplastics (MPs) in marine environments simultaneously affect microalgae with UV-B radiation, while their joint effecting mechanisms remain largely unknown. To fill this research gap, the joint effects of polymethyl methacrylate (PMMA) MPs and UV-B radiation (natural environments intensity) on the model marine diatom Thalassiosira pseudonana were investigated. Antagonism was found between the two factors with regards to population growth. Furthermore, we found more inhibited population growth and photosynthetic parameters when pre-treated with PMMA MPs compared to pre-treated with UV-B radiation before joint-treated by the two factors. Transcriptional analysis elucidated that UV-B radiation could alleviate the down-regulation of photosynthetic (PSII, cyt b6/f complex and photosynthetic electron transport) and chlorophyll biosynthesis genes caused by PMMA MPs. Besides, the genes encoding carbon fixation and metabolisms was up-regulated under UV-B radiation, which could provide extra energy for the enhanced anti-oxidative activities and DNA replication-repair processes. These consequences showed that the toxicity of PMMA MPs was comprehensively alleviated when T. pseudonana was jointed treated by UV-B radiation. Our results reveled the underlying molecular mechanisms of antagonistic effects between PMMA MPs and UV-B radiation. This study provides important information that environmental factors like UV-B radiation should be considered when accessing the ecological risks of MPs on marine organisms.

The adverse impact of microplastics and their attached pathogen on hemocyte function and antioxidative response in the mussel Mytilus galloprovincialis.

Microplastics (MPs) are widely distributed in marine environments, and they are easily attached by various microorganisms, including pathogenic bacteria. When bivalves mistakenly eat MPs, pathogenic bacteria attached to MPs enter their bodies through the Trojan horse effect, causing adverse effects. In this study, the mussel Mytilus galloprovincialis was exposed to aged polymethylmethacrylate MPs (PMMA-MPs, 20 µm) and Vibrio parahaemolyticus attached to PMMA-MPs to explore the effect of synergistic exposure by measuring lysosomal membrane stability, ROS content, phagocytosis, apoptosis in hemocytes, antioxidative enzyme activities and apoptosis-related gene expression in gills and digestive glands. The results showed that MP exposure alone did not cause significant oxidative stress in mussels, but after long-term coexposure to MPs and V. parahaemolyticus, the activities of antioxidant enzymes were significantly inhibited in the gills of mussels. Both single MP exposure and coexposure will affect hemocyte function. Coexposure can induce hemocytes to produce higher ROS, improve phagocytosis, significantly reduce the stability of the lysosome membrane, and induce the expression of apoptosis-related genes, causing apoptosis of hemocytes compared with single MP exposure. Our results demonstrate that MPs attached to pathogenic bacteria have stronger toxic effects on mussels, which also suggests that MPs with pathogenic bacteria might have an influence on the immune system and cause disease in mollusks. Thus, MPs may mediate the transmission of pathogens in marine environments, posing a threat to marine animals and human health. This study provides a scientific basis for the ecological risk assessment of MP pollution in marine environments.

Understanding the aggregation, consumption, distribution and accumulation of nanoparticles of polyvinyl chloride and polymethyl methacrylate in Ruditapes philippinarum.

Plastic products have become an integral part of our life. A widespread usage, high stability, uncontrolled disposal and slow degradation of plastics in the environment led to the generation and accumulation of nanoparticles of polymers (NPs) in the marine environment. However, little is known about the aggregation, consumption and distribution of NPs from common polymers such as polyvinyl chloride (NP-PVC) and polymethyl methacrylate (NP-PMMA) inside marine animal physiologies. In the current study, two types of polymers (PVC and PMMA) × four exposure concentrations (1, 5, 15 and 25 mg/L) × four times (4, 8, 12 and 24 h) exposure studies were conducted to understand the consumption and distribution of luminescent NP-PVC (98.6 ± 17.6 nm) and NP-PMMA (111.9 ± 37.1 nm) in R. philippinarum. Under laboratory conditions, NP-PVC showed a higher aggregation rate than NP-PMMA in seawater within a period of 24 h. Aggregations of NPs increased with an increase in initial NP concentrations, leading to significant settling of nanoparticles within 24 h exposure. Such aggregation and settling of particles enhanced the consumption of NPs by benthic filter-feeding R. philippinarum at all exposure concentrations during 4 h exposure. More interestingly, NP-PVC and NP-PMMA were observed in large amounts in both liver and gills (22.6 % - 29.1 %) of the clams. Furthermore, NP-PVC was detected in most organs of R. philippinarum as compared to NP-PMMA. This study demonstrates that different polymers distribute and accumulate differently in the same biological model under laboratory exposure conditions based on their chemical nature.

Novel Tuning of PMMA Orthopedic Bone Cement Using TBB Initiator: Effect of Bone Cement Extracts on Bioactivity of Osteoblasts and Osteoclasts.

Bone cement containing benzoyl peroxide (BPO) as a polymerization initiator are commonly used to fix orthopedic metal implants. However, toxic complications caused by bone cement are a clinically significant problem. Poly (methyl methacrylate) tri-n-butylborane (PMMA-TBB), a newly developed material containing TBB as a polymerization initiator, was found to be more biocompatible than conventional PMMA-BPO bone cements due to reduced free radical generation during polymerization. However, free radicals might not be the only determinant of cytotoxicity. Here, we evaluated the response and functional phenotypes of cells exposed to extracts derived from different bone cements. Bone cement extracts were prepared from two commercial PMMA-BPO cements and an experimental PMMA-TBB. Rat bone marrow-derived osteoblasts and osteoclasts were cultured in a medium supplemented with bone cement extracts. More osteoblasts survived and attached to the culture dish with PMMA-TBB extract than in the culture with PMMA-BPO extracts. Osteoblast proliferation and differentiation were higher in the culture with PMMA-TBB extract. The number of TRAP-positive multinucleated cells was significantly lower in the culture with PMMA-TBB extract. There was no difference in osteoclast-related gene expression in response to different bone cement extracts. In conclusion, PMMA-TBB extract was less toxic to osteoblasts than PMMA-BPO extracts. Although extracts from the different cement types did not affect osteoclast function, PMMA-TBB extract seemed to reduce osteoclastogenesis, a possible further advantage of PMMA-TBB cement. These implied that the reduced radical generation during polymerization is not the only determinant for the improved biocompatibility of PMMA-TBB and that the post-polymerization chemical elution may also be important.

Effects of nanoplastics on zebrafish embryo-larval stages: A case study with polystyrene (PS) and polymethylmethacrylate (PMMA) particles.

Plastic production has been rising consistently in the last 30 years and with it, the presence of plastic particles in the environment. A decrease in size often increases the bioavailability and reactivity of the particles. In this study the impact of polystyrene (PS; 22 nm) and polymethylmethacrylate (PMMA; 32 nm) nanoparticles on zebrafish embryo-larval stages was assessed by studying mortality, hatching, morphological features, and biochemical endpoints (associated with neurotransmission, antioxidant status and oxidative damage, and energy metabolism) after 96 h exposure, and swimming behavior after 120 h exposure. Organisms exposed to PMMA nanoparticles exhibited higher mortality and pericardial edema than those exposed to PS nanoparticles but displayed less effects on swimming behavior. Biochemical endpoints were altered to a higher degree in organisms exposed to PS nanoparticles (acetylcholinesterase, glutathione S-transferase and catalase activities) but higher peroxidative damage was found after exposure to lower concentrations of PMMA nanoparticles. Both types of nanoparticles affected energy metabolism with higher levels of glycogen found in animals exposed to PS nanoparticles. The use of integrated biomarker response index confirmed that PS nanoparticles had a higher impact on biochemical endpoints of zebrafish.

A transparent hydrophilic anti-biofouling coating for intraocular lens materials prepared by "bridging" of the intermediate adhesive layer.

The attachment of bio-foulants, including unwanted cells, proteins, and bacteria, to a medical device such as an intraocular lens can lead to implantation failure. Hydrophilic polymers are often used as surface modifiers in the fabrication of anti-biofouling coatings, but a hydrophilic coating can easily become swollen and peel off the substrate. In this study, we chose polymethyl methacrylate (PMMA) as the representative material of intraocular lenses because PMMA has better biocompatibility, a higher refractive index, better optical clarity, lighter weight, more stable performance, and lower cost than other intraocular lens materials. We fabricated polyvinyl alcohol (PVA) coatings with or without a "bridge", that is, an intermediate adhesive layer (AL), to increase the adhesion bonding effect between the anti-biofouling coating and the substrate. The results indicated that the prepared coatings were transparent and noncytotoxic. Moreover, the anti-adhesion properties of the cells and the resistance properties to nonspecific protein adsorption of PMMA modified by both AL and PVA coatings were better and more durable compared with the sample only modified with a physically dipped PVA coating. The coating prepared by AL "bridging" provides a new strategy for the preparation of a transparent hydrophilic anti-biofouling coating suitable for PMMA intraocular lens materials.

Glutathione and Reactive Oxygen Species Dual-Responsive Block Copolymer Prodrugs for Boosting Tumor Site-Specific Drug Release and Enhanced Antitumor Efficacy.

A remarkable hallmark of cancer cells is the heterogeneous coexistence of overproduced intracellular glutathione (GSH) and a high level of reactive oxygen species (ROS) compared with those in normal cells, which have been frequently used as the stimuli to trigger drug release from the nanocarriers. Most of the stimuli-responsive delivery vehicles have been designed to respond to only one redox stimulus (e.g., GSH or ROS). Herein, we develop a GSH and ROS dual-responsive amphiphilic diblock copolymer prodrug (BCP) (GR-BCP) consisting of poly(ethylene glycol) (PEG)- and camptothecin (CPT)-conjugated poly(methacrylate) in the side chains via thioether bonds. In comparison, GSH or ROS single-responsive BCPs (G-BCPs or R-BCPs) were prepared, where CPT drugs were linked by disulfide or thioketal bonds, respectively. The three BCPs can form well-defined spherical micellar nanoparticles in an aqueous solution with a diameter of ∼50 nm. Compared with G-BCP and R-BCP, GR-BCP realized the highest cytotoxicity against HeLa cells with the half-inhibitory concentration (IC50) of 6.3 µM, which is much lower than 17.8 µM for G-BCP and 28.9 µM for R-BCP. Moreover, for in vivo antitumor performance, G-BCP, R-BCP, and GR-BCP showed similar efficiencies in blood circulation and tumor accumulation after intravenous injection. However, GR-BCP realized the most efficient tumor suppression with few side effects. Our findings demonstrate that intracellular GSH and ROS dual-responsive BCPs show a more efficient responsive drug release inside tumor cells for boosting the antitumor efficacy as compared with GSH or ROS single-responsive BCPs, which provides novel strategies for designing redox-responsive BCPs.

Effects of Molar Ratios of Two Immiscible Monomers toward Development of an Amphiphilic, Highly Stretchable, Bioadhesive, Self-Healing Copolymeric Hydrogel and its Mineral-Active Cellular Behavior.

Here, we report the striking properties such as high stretchability, self-healing, and adhesiveness of an amphiphilic copolymeric hydrogel (poly(acrylic acid)-poly(methyl methacrylate) (PAA-PMMA) gel) synthesized from two immiscible monomers-acrylic acid (AA) and methyl methacrylate (MMA)-through a simple free radical polymerization in an aqueous medium. The developed hydrogel, with a specific molar ratio of MMA and AA, is self-healable, which is attributed to the hydrophobic interaction arising from methyl groups of PMMA, as well as the breakdown and reformation of sacrificial noncovalent cross-linking through the weak hydrogen bonds between the carboxylic acid groups of PAA and methoxy groups of PMMA. The energy dissipation values in the hysteresis test signify the excellent self-recoverability of the hydrogel. The developed hydrogel showed adhesive behavior to the surfaces of polystyrene, glass, wood, metal, stone, ceramics, pork skin, and human skin. The physical and mechanical properties of the PAA-PMMA gel were fine-tuned through changes in the MMA/AA ratio and pH. Moreover, the PAA-PMMA hydrogel can serve as a template for calcium phosphate mineralization to yield a hydrogel composite, which improved MC3T3 cell adhesion and proliferation. Overall, we propose that depending on synthesis parameters and other scenarios, the synthesized PAA-PMMA hydrogel could potentially be employed in varying biomedical and industrial applications.

Electrophoretic deposition of polymethylmethacrylate and composites for biomedical applications.

For the first time, an electrophoretic deposition (EPD) method has been developed for the deposition of polymethylmethacrylate (PMMA) and PMMA-alumina films for biomedical implant applications. The proposed biomimetic approach was based on the use of a bile salt, sodium cholate (NaCh), which served as a multifunctional solubilizing, charging, dispersing and film-forming agent. Investigations revealed PMMA-Ch- and PMMA-alumina interactions, which facilitated the deposition of PMMA and PMMA-alumina films. This approach allows for the use of a non-toxic water-ethanol solvent for PMMA. The proposed deposition strategy can also be used for co-deposition of PMMA with other functional materials. The PMMA and composite films were tested for biomedical implant applications. The PMMA-alumina films showed statistically improved metabolic results compared to both the bare stainless steel substrate and pure PMMA films. Alkaline phosphatase (ALP) activity affirmed the bioactivity and osteoconductive potential of PMMA and composite films. PMMA-alumina films showed greater ALP activity than both the PMMA-coated and uncoated stainless steel.

Combination Antibiotic Delivery in PMMA Provides Sustained Broad-Spectrum Antimicrobial Activity and Allows for Postimplantation Refilling.

Antibiotics are commonly added to poly(methyl methacrylate) (PMMA) by surgeons to locally treat infections such as in bone cement for joint replacement surgeries, as well as implantable antimicrobial "beads". However, this strategy is of limited value in high-risk patients where infections can be recurrent or chronic and otherwise hard to treat. Also, when only one drug is incorporated and applied toward polymicrobial infections (multiple bacterial species), there is a high risk that bacteria can develop antibiotic resistance. To combat these limitations, we developed a combination antibiotic PMMA composite system composed of rifampicin-filled ß-cyclodextrin (ß-CD) microparticles added into PMMA filled with a second drug. Different formulations were evaluated through zone of inhibition, drug activity, antibiotic release, and refilling, as well as mechanical studies. Our combination antibiotic PMMA composite system achieved up to an 8-fold increase in the duration of antimicrobial activity in comparison to clinically used antibiotic-filled PMMA. Inclusion of CD microparticles also allowed for refilling of additional antibiotics after simulated implantation, resulting in additional windows of therapeutic efficacy. Mechanical testing showed that our tested formulations did have a small, but significant decrease in mechanical properties when compared to unmodified controls. While further studies are needed to determine whether the tested formulations are still suitable for load-bearing applications (e.g., bone cement), our composites are certainly amenable for a variety of nonload-bearing applications (e.g., antimicrobial "beads" and temporary spacer in two-stage arthroscopic revisions).

Targeted delivery of poly (methyl methacrylate) particles in colon cancer cells selectively attenuates cancer cell proliferation.

Poly (methyl methacrylate) (PMMA) is basically biocompatible polyester with high resistance to chemical hydrolysis, and high drug permeability and the most important characteristics of PMMA is that it does not produce any toxicity. There is not much information about PMMA action on the colon cancer cells. In the present study, we have synthesized PMMA nanoparticles. The distribution pattern of PMMA particles was analysed by Zeta sizer and the size of the particles was calculated by using quasi elastic light scattering (QELS). The surface structure and the morphology of PMMA were characterized by transmission electron microscope (TEM) and scanning electron microscope (SEM), respectively. We have also analysed their effects on cancerous cells (human colorectal carcinoma cells, HCT-116) and normal, healthy cells (human embryonic kidney cells, HEK-293) by using morphometric, MTT, DAPI and wound healing methods. We report that PMMA particles inhibited the cancer cell viability in a dose-dependent manner. The lower dose (1.0 µg/ml) showed a moderate decrease in cancer cell viability, whereas higher dosages (2.5 µg/ml, 5.0 µg/mL and 7.5 µg/mL) showed steadily decrease in the cancer cell viability. We also report that PMMA is highly selective to cancerous cells (HCT-116), as we did not find any action on the normal healthy cells (HEK-293). In conclusion, our results suggest PMMA particles are potential biomaterials to be used in the treatment of colon cancer.

Toxicity screening of a novel poly(methylmethacrylate)-Eudragit nanocarrier on L929 fibroblasts.

Translation of innovative drug delivery nanosystems into the market involves an early toxicity screening in the development phase. Previously, we showed that inclusion of the polymer Eudragit (EUD) into poly(methylmethacrylate) (PMMA) nanoparticles (NP) resulted in a novel nanocarrier (PMMA-EUD) with an improved biomedical performance. The safety of this novel nanoparticulate system (PMMA-EUDNPs) was assessed in this work and compared to that of the original PMMANPs by using an integrated approach, comprising in vitro toxicity assessment and NPs physicochemical characterization in water and cell medium. For toxicity assessment several endpoints were analysed, including cell death, oxidative stress, and genotoxicity using L929 fibroblasts. PMMA-EUDNPs proved to be more hydrophobic than the original PMMANPs. Also, charge of both NPs was strongly affected by cell medium. On the other hand, the novel nanosystem was easily uptaken by L929 cells and did not display relevant in vitro cytotoxic or genotoxic effects. On the contrary, PMMANPs were less internalized in cells and proved to be genotoxic, as measured by the micronucleus assay. To conclude, our results provide initial evidence about the safety of the novel and promising PMMA-EUD nanoparticulate system, enabling its further development towards applications for drug delivery.

Multimeric System of RGD-Grafted PMMA-Nanoparticles as a Targeted Drug- Delivery System for Paclitaxel.

BACKGROUND: Polymer-based nanoparticles as drug-delivery systems offer new therapeutic opportunities. Among them, ligand-mediated targeting, which increases selectivity and efficacy, allows controllable drug delivery. The aim of the this research was to prepare and characterize poly(methyl methacrylate) (PMMA) nanoparticles grafted with the -Arginine, Glycine, Aspartic Acid (RGD)- peptide sequence as a promising smart drug delivery system for Paclitaxel (PTX), directed at the sites of integrin receptor overexpression. METHODS: Nanoparticles were characterized by FT-IR and Raman spectroscopy, dynamic light scattering, zeta potential and transmission electron microscopy. RESULTS: RGD-PMMA-PTX size distribution was 17.58 ± 7.45 nm with a zeta potential of -38.73 ± 5.62 mV. According to the boxLucas Model, PTX was incorporated into nanoparticles with an entrapment efficiency of 100% (evaluated by HPLC analysis). In vitro sustained release was determined, with the maximum release of 55% and 40% after 21 days at pH 5.3 and 7.4, respectively. The highest inhibition on cell proliferation was found with RGD-PMMA-PTX nanoparticles (90 %). CONCLUSION: The obtained results showed that RGD-PMMA-PTX represents an attractive and suitable therapeutic approach for targeting overexpressed integrins in the cancer cells.

Effect of surface sealant agents on the surface roughness and color stability of denture base materials.

STATEMENT OF PROBLEM: The effects of surface sealant agents on the surface roughness and color stability of denture base materials are unknown. PURPOSE: The purpose of this in vitro study was to evaluate the effects of different polishing methods on the surface roughness and color stability of denture base materials. MATERIAL AND METHODS: A total of 120 specimens were fabricated from 2 poly(methyl methacrylate) (PMMA) and 1 polyamide denture base materials and divided into 4 groups (n=10 in each group) according to the applied surface treatment procedure: conventional polishing (control) and 3 surface sealant coupling methods. Surface roughness average (Ra) values were measured using a profilometer. Color parameters were measured using a spectrophotometer before and after being stained with coffee. Color differences (CIEDE 2000 [ΔE00]) were calculated. Data were statistically analyzed using 2-way ANOVA and Tukey honest significant difference test (α=.05). RESULTS: No statistically significant difference were found between surface roughness values of the control and those of the specimens treated using a surface sealant agent (P>.05). The highest color difference was calculated for the polyamide control group. Statistically significant differences were found between the control group and the group treated with the polyamide surface sealant agent (P<.05). CONCLUSIONS: All specimens had a surface roughness value higher than the plaque accumulation threshold (0.20 µm). The color changes observed were clinically unacceptable, except for conventionally polished and one type of surface sealant applied microwave polymerized PMMA denture base material.

Controlling fungal biofilms with functional drug delivery denture biomaterials.

Candida-associated denture stomatitis (CADS), caused by colonization and biofilm-formation of Candida species on denture surfaces, is a significant clinical concern. We show here that modification of conventional denture materials with functional groups can significantly increase drug binding capacity and control drug release rate of the resulting denture materials for potentially managing CADS. In our approach, poly(methyl methacrylate) (PMMA)-based denture resins were surface grafted with three kinds of polymers, poly(1-vinyl-2-pyrrolidinone) (PNVP), poly(methacrylic acid) (PMAA), and poly(2-hydroxyethyl methacrylate) (PHEMA), through plasma-initiated grafting polymerization. With a grafting yield as low as 2 wt%, the three classes of new functionalized denture materials showed significantly higher drug binding capacities toward miconazole, a widely used antifungal drug, than the original PMMA denture resin control, leading to sustained drug release and potent biofilm-controlling effects against Candida. Among the three classes of functionalized denture materials, PNVP-grafted resin provided the highest miconazole binding capability and the most powerful antifungal and biofilm-controlling activities. Drug binding mechanisms were studied. These results demonstrated the importance of specific interactions between drug molecules and functional groups on biomaterials, shedding lights on future design of CADS-managing denture materials and other related devices for controlled drug delivery.

New method for viewing Krehbiel flow by polymethylmethacrylate particles suspended in fluorescein solution.

PURPOSE: To investigate the changes in the tear flow velocities caused by ageing. METHODS: Ninety-nine subjects (41 men, mean age 48.3 ± 20.7 years) were recruited from the Department of Ophthalmology of the Ehime University Hospital. None of the subjects had serious abnormalities of the external surface of the eye. The Krehbiel flow of tears was determined by 40-µm polymethylmethacrylate (PMMA) beads suspended in a fluorescein sodium solution (PPF). The movement of the beads was video recorded through a slit-lamp during normal blinking. The flow of the beads was determined with a Motion ANALYZER(®) software (KEYENCE Co., Osaka, Japan). The velocity of the beads in young age, 20-40 years, middle age, 41-60 years and old age, ≥61 years, groups was determined. RESULTS: The equation describing the velocity (mm/second) of the PMMA particles as a function of age in the lower tear meniscus measured in the direction of the lacrimal punctum was Y = 2.49-0.04X, where Y = velocity and X = age (r(2) = 0.214; p < 0.0001). For the upper meniscus, the equation was Y = 4.83-0.05X (r(2) = 0.195, p < 0.0001). The average velocity was 0.70 ± 1.66 mm/second in the lower and 2.16 ± 1.93 mm/second in the upper tear meniscus (p < 0.0001). The particle velocity decreased significantly with increasing age, but no significant difference between the male and female groups except for the lower tear meniscus when all subjects were analysed. CONCLUSION: The PPF technique is a simple method of examining Krehbiel flow of tears and may be used for evaluating functional nasolacrimal duct obstruction quantitatively.

An efficient and reproducible method to culture Bergmann and cortical radial glia using textured PMMA.

BACKGROUND: Radial glia cells comprise the principal population of neural stem cells (NSC) during development. Attempts to develop reproducible radial glia and NSC culture methods have met with variable results, yielding non-adherent cultures or requiring the addition of growth factors. Recent studies demonstrated that a 2-µm patterned poly-methyl methacrylate (ln2 PMMA) grooved scaffold, by mimicking the biophysical and microtopographic properties of the embryonic NSC niche, induces the de-differentiation of glial cells into functional radial glia cells. NEW METHOD: Here we describe a method for obtaining cultures of adherent Bergmann radial glia (BRG) and cortical radial glia (CRG). The growth substrate is ln2 PMMA and the addition of growth factors is not required. RESULTS: Postnatal glia obtained from mouse cerebellum or cerebral cortex and grown on ln2 PMMA adopted a BRG/CRG phenotype characterized by a bipolar shape, the up-regulation of progenitor markers such as nestin and Sox2, and the down-regulation of vimentin and GFAP. Neurons cultured over the BRG/CRG aligned their processes with those of the glial shafts, thus mimicking the behavior of migrating neuronal cells. COMPARISON WITH EXISTING METHODS: The ln2 PMMA culture method offers an ideal system for analyzing both the biochemical factors controlling the neurogenic potential of BRG/CRG and neuronal migration. CONCLUSIONS: The ln2 PMMA method is a reproducible system to obtain immature BRG/CRG preparations in vitro. It can be used to study the properties of CNS progenitor cells as well as the interactions between radial glia and neurons, and supports cultured progenitors for use in different applications.

Sol-gel approach for fabrication of coated anodized titanium wire for solid-phase microextraction: highly efficient adsorbents for enrichment of trace polar analytes.

Nanotubular titania film was prepared in situ on titanium wire and was used as the fiber substrate for solid-phase microextraction (SPME) because of its high surface-to-volume ratio, easy preparation, and mechanical stability. Three different functional coatings, ß-cyclodextrin (ß-CD), ß-cyclodextrin-co-poly(ethylenepropylene glycol) (ß-CD/PEG), and polyethylene glycol (PEG)-based sorbents were chemically bonded to the nanostructured wire surface via sol-gel technology to further enhance the absorbing capability and extraction selectivity. Coupled to gas chromatography-flame ionic detection (GC-FID), the prepared SPME fibers were investigated using diverse compounds. The results indicated that the fibers showed good mechanical strength, excellent thermal stability, and wonderful capacity and selectivity to polar compounds, including polar aromatic compounds, alcohols, and ketones. Combining the superior hydrophilic property of a bonded functional molecule and the highly porous structure of a fiber coating, the prepared PEG-coated SPME fiber showed much higher adsorption affinity to ephedrine and methylephedrine than ß-CD and ß-CD/PEG fibers. The as-established PEG-coated SPME-GC analytical method provided excellent sensitivity (LODs, 0.004 and 0.001 ng mL(-1) for ephedrine and methylephedrine, respectively) and better linear range (0.01-2 000 µg L(-1)). In addition, it has surprising repeatability and reproducibility. Finally, the present approach was used to analyze ephedrine and methylephedrine from real urine samples, and reliable results were obtained.