Bioinert Control of Zwitterionic Poly(ethylene terephtalate) Fibrous Membranes.

Poly(ethylene terephtalate) (PET)-based materials face general biofouling issues that we addressed by grafting a copolymer of glycidyl methacrylate and sulfobetaine methacrylate, poly(GMA- r-SBMA). The grafting procedure involved a dip-coating step followed by UV-exposure and led to successful grafting of the copolymer as evidenced by X-ray photoelectron spectroscopy and zeta potential measurements. It did not modify the pore size nor the porosity of the PET membranes. In addition, their surface hydrophilicity was considerably improved, with a water contact angle falling to 30° in less than 20 s and 0° in less than 1 min. The effect of copolymer concentration in the coating bath (dip-coating procedure) and UV exposure time (UV step) were scrutinized during biofouling studies involving several bacteria such as Escherichia coli and Stenotrophomonas maltophilia, but also whole blood and HT1080 fibroblasts cells. The results indicate that if all conditions led to improved biofouling mitigation, due to the efficiency of the zwitterionic copolymer and grafting procedure, a higher concentration (15 mg/mL) and longer UV exposure time (at least 10 min) enhanced the grafting density which reflected on the biofouling results and permitted a better general biofouling control regardless of the nature of the biofoulant (bacteria, blood cells, fibroblasts).

A Zwitterionic-Shielded Carrier with pH-Modulated Reversible Self-Assembly for Gene Transfection.

Cationic vectors are ideal candidates for gene delivery thanks to their capability to carry large gene inserts and their scalable production. However, their cationic density gives rise to high cytotoxicity. We present the proper designed core-shell polyplexes made of either poly(ethylene imine) (PEI) or poly(2-dimethylamino ethyl methacrylate) (PDMAEMA) as the core and zwitterionic poly(acrylic acid)-block-poly(sulfobetaine methacrylate) (PAA-b-PSBMA) diblock copolymer as the shell. Gel retardation and ethidium bromide displacement assays were used to determine the PEI/DNA or PDMAEMA/DNA complexation. At neutral pH, the copolymer serves as a protective shell of the complex. As PSBMA is a nonfouling block, the shell reduced the cytotoxicity and enhanced the hemocompatibility (lower hemolysis activity, longer plasma clotting time) of the gene carriers. PAA segments in the copolymer impart pH sensitivity by allowing deshielding of the core in acidic solution. Therefore, the transfection efficiency of polyplexes at pH 6.5 was better than at pH 7.0, from ß-galactosidase assay, and for all PAA-b-PSBMA tested. These results were supported by more favorable physicochemical properties in acidic solution (zeta potential, particle size, and interactions between the polymer and DNA). Thus, the results of this study offer a potential route to the development of efficient and nontoxic pH-sensitive gene carriers.

Dopamine-Induced Surface Zwitterionization of Expanded Poly(tetrafluoroethylene) for Constructing Thermostable Bioinert Materials.

Although energy-demanding, the surface modification of polytetrafluoroethylene (PTFE) for biomedical applications is mandatory to mitigate irreversible biofouling that occurs whenever PTFE comes into contact with biological fluids. Here, we propose to take advantage of the adhesive properties of dopamine (DA) and of the antifouling ability of various zwitterionic monomers (sulfobetaine methacrylate (SBMA), sulfobetaine methacrylamide (SBAA), sulfobetaine acrylamide (SBAA'), and 4-vinylpyridine propylsulfobetaine (4VPPS)) and form antifouling coatings by copolymerization on the surface of expanded PTFE membranes. This simple, low-energy, and one-step coating procedure arises in significant biofouling mitigation. All zwitterionic coatings led to important reduction of biofouling by red blood cell conentrate (88-94%), platelet conentrate (70-90%), whole blood (40-66%), or bacteria (83-96%). Also, it is shown that the interactions of polydopamine with ePTFE are stable even at high temperatures. However, the zwitterionic monomers are differently affected. While the performance of SBMA coatings decreased (as SBMA is prone to hydrolysis), those of SBAA, SBAA', and 4VPPS coatings were generally maintained. All in all, this study illustrates that efficient and stable antifouling zwitterionic coatings can be generated onto PTFE membranes for biomedical applications, without the use of conventional high-energy-demanding surface modification processes.

Microfluidic colorimetric detection platform with sliding hybrid PMMA/paper microchip for human urine and blood sample analysis.

A microfluidic colorimetric detection (MCD) platform consisting of a sliding hybrid PMMA/paper microchip and a smart analysis system is proposed for the convenient, low-cost and rapid analysis of human urine and whole blood samples. The sliding PMMA/paper microchip comprises a PMMA microfluidic chip for sample injection and transportation, a paper strip for sample filtration (urine) or separation (blood), and a sealed paper-chip detection zone for sample reaction and detection. In the proposed device, the paper-chip is coated with bicinchoninic acid (BCA) and biuret reagent and is then assembled into the PMMA microchip and packaged in aluminum housing. In the detection process, the PMMA/paper microchip is slid partially out of the housing, and 2 µL of sample (urine or whole blood) is dripped onto the sample injection zone. The chip is then slid back into the housing and the sample is filtered/separated by the paper strip and transferred under the effects of capillary action to the sealed paper-chip detection zone. The housing is inserted into the color analysis system and heated at 45 °C for 5 min to produce a purple-colored reaction complex. The complex is imaged using a CCD camera and the RGB color intensity of the image is then analyzed using a smartphone to determine the total protein (TP) concentration of the sample. The effectiveness of the proposed method is demonstrated using TP control samples with known concentrations in the range of 0.03-5.0 g/dL. The detection results obtained for 50 human urine samples obtained from random volunteers are shown to be consistent with those obtained from a conventional hospital analysis system (R2 = 0.992). Moreover, the detection results obtained for the albumin (ALB) and creatine (CRE) concentrations of 50 whole blood samples are also shown to be in good agreement with the results obtained from the hospital analysis system (R2 = 0.982 and 0.988, respectively).

Proteomic analysis provides insights on venom processing in Conus textile.

Conus species of marine snails deliver a potent collection of toxins from the venom duct via a long proboscis attached to a harpoon tooth. Conotoxins are known to possess powerful neurological effects and some have been developed for therapeutic uses. Using mass-spectrometry based proteomics, qualitative and quantitative differences in conotoxin components were found in the proximal, central and distal sections of the Conus textile venom duct suggesting specialization of duct sections for biosynthesis of particular conotoxins. Reversed phase HPLC followed by Orbitrap mass spectrometry and data analysis using SEQUEST and ProLuCID identified 31 conotoxin sequences and 25 post-translational modification (PTM) variants with King-Kong 2 peptide being the most abundant. Several previously unreported variants of known conopeptides were found and this is the first time that HyVal is reported for a disulfide rich Conus peptide. Differential expression along the venom duct, production of PTM variants, alternative proteolytic cleavage sites, and venom processing enroute to the proboscis all appear to contribute to enriching the combinatorial pool of conopeptides and producing the appropriate formulation for a particular hunting situation. The complementary tools of mass spectrometry-based proteomics and molecular biology can greatly accelerate the discovery of Conus peptides and provide insights on envenomation and other biological strategies of cone snails.

Surface Zwitterionization of Expanded Poly(tetrafluoroethylene) via Dopamine-Assisted Consecutive Immersion Coating.

Expanded polytetrafluoroethylene (ePTFE) is one of the materials widely used in the biomedical field, yet its application is being limited by adverse reactions such as thrombosis when it comes in contact with blood. Thus, a simple and robust way to modify ePTFE to be biologically inert is sought after. Modification of ePTFE without high-energy pretreatment, such as immersion coating, has been of interest to researchers for its straightforward process and ease in scaling up. In this study, we utilized a two-step immersion coating to zwitterionize ePTFE membranes. The first coating consists of the co-deposition of polyethylenimine (PEI) and polydopamine (PDA) to produce amine groups in the surface of the ePTFE for further functionalization. These amine groups from PEI will be coupled with the epoxide group of the zwitterionic copolymer, poly(GMA-co-SBMA) (PGS), via a ring-opening reaction in the second coating. The coated ePTFE membranes were physically and chemically characterized to ensure that each step of the coating is successful. The membranes were also tested for their thrombogenicity via quantification of the blood cells attached to it during contact with biological solutions. The coated membranes exhibited around 90% reduction in attachment with respect to the uncoated ePTFE for both Gram-positive and Gram-negative strains of bacteria (Staphylococcus aureus and Escherichia coli). The coating was also able to resist blood cell attachment from human whole blood by 81.57% and resist red blood cell attachment from red blood cell concentrate by 93.4%. These ePTFE membranes, which are coated by a simple immersion coating, show significant enhancement of the biocompatibility of the membranes, which shows promise for future use in biological devices.

Multifunctional microchip-based distillation apparatus II - Aerated distillation for sulfur dioxide detection.

A multifunctional microchip-based distillation apparatus is presented for the distilled of sulfur dioxide (SO2) in food products. The microchip is fabricated on poly(methyl methacrylate) (PMMA) substrates, and comprises a sample zone, a buffer zone, a serpentine distillation column, and a collection zone. In the process, the sample is introduced into the sample zone and is heated under carefully controlled temperature and time conditions. The resulting SO2 and water vapor are carried by nitrogen (N2) gas to the distillation column, where the SO2 is separated from the water vapor via the condensing effects of a continuous cold water flow. Finally, the SO2 is transported to the collection zone, where it is collected with hydrogen peroxide (H2O2) and its concentration determined using an alkali-based titration and paper-based detection method. A distillation efficiency of 90.5% is obtained under the optimal distillation conditions at concentrations of 20-4000 ppm. Moreover, a linear correlation (R2 = 0.9997) is observed between the experimental measurements of the SO2 concentration and the known concentration. The validity of the presented microchip-based distillation apparatus is further investigated by distilling the SO2 concentrations of 25 commodity samples. The detection results show that the deviation does not exceed 5.4% compared with the traditional official method.