4-META/MMA-TBB resin containing nano hydroxyapatite induces the healing of periodontal tissue repair in perforations at the pulp chamber floor.

We aimed to evaluate the materials based on 4-methacryloxyethyl trimellitate anhydride/methyl methacrylate tri-n-butylborane (Super-bond [SB]) and nano hydroxyapatite (naHAp) for the repair of perforation at pulp chamber floor (PPF) in vitro and in vivo models. SB and naHAp were mixed in the mass ratio of 10% or 30% to produce naHAp/SB. Human periodontal ligament stem cells (HPDLSCs) were cultured on resin discs of SB or naHAp/SB to analyze the effects of naHAp/SB on cell adhesion, proliferation, and cementoblastic differentiation. A rat PPF model was treated with SB or naHAp/SB to examine the effects of naHAp/SB on the healing of defected cementum and periodontal ligament (PDL) at the site of PPF. HPDLSCs were spindle-shaped and adhered to all resin discs. Changing the resin from SB to naHAp/SB did not significantly alter cell proliferation. Both 10% and 30% naHAp/SB were more effective than SB in promoting cementoblastic differentiation of HPDLSCs. In the rat PPF model, 30% naHAp/SB was more effective than SB in promoting the formation Sharpey's fiber-like structures with expression of the PDL-related marker and cementum-like structures with expression of cementum-related markers. In conclusion, 30% naHAp/SB can be the new restorative material for PPF because it exhibited the abilities of adhering to dentin and healing of defected periodontal tissue.

Extending the Affinity Range of Weak Affinity Chromatography for the Identification of Weak Ligands Targeting Membrane Proteins.

The identification of weak-affinity ligands targeting membrane proteins is of great interest in Fragment-Based Drug Design (FBDD). Recently, miniaturized weak affinity chromatography (WAC) has been proposed as a valuable tool to study interactions between small ligands and wild-type membrane proteins embedded in so-called nanodisc biomimetic membranes immobilized on GMA-co-EDMA monoliths in situ-synthesized in capillary columns (less than one microliter in volume). In this proof-of-concept study, the achievable affinity range was limited to medium affinity (low micromolar range). The present work investigates different strategies to extend the affinity range towards low affinities, either by increasing the density of membrane proteins on the chromatographic support or by reducing non-specific interactions with the monolith. The combination of the use of a new and more hydrophilic monolithic support (poly(DHPMA-co-MBA)) and a multilayer nanodisc grafting process (up to three layers) allows a significant increase in the membrane protein density by a more than three-fold factor (up to 5.4 pmol cm-1). Such an increase in protein density associated with reduced non-specific interactions makes it possible to extend the range of detectable affinity, as demonstrated by the identification and characterization of affinities of very low-affinity ligands (Kd values of several hundred micromolar) for the adenosine receptor AA2AR used as a model protein, which was not possible before. The affinity was confirmed by competition experiments.

Effects of methacryloyloxydecyl dihydrogen phosphate on bonding of tri-n-butylborane-initiated resin to human enamel.

This study investigated the effect of 10-methacryloyloxydecyl dihydrogen phosphate (MDP) in methyl methacrylate (MMA) monomer on the adhesion of tri-n-butylborane (TBB)-initiated resins (MDP/MMA-TBB resins) to human enamel. Enamel surface conditions were either polished only or phosphatized surfaces. The 1.0, 1.7, and 2.0 mol% MDP/MMA-TBB resins, 4-methacryloxyethyl trimellitate anhydride (4-META)/MMA-TBB resin and MMA-TBB resin were prepared as luting materials. The shear bond strength was determined before and after thermocycles, and the results were compared using non-parametric statistical analyses (each, n=15). The MDP/MMA-TBB resins showed significantly better bond durability to enamel than other resins with or without etching. The 1.7 and 2.0 mol% MDP/MMA-TBB resins were suggested to be the optimum MDP concentrations from pre- and post-thermocycling results for the non-etched specimens. The TBB initiator resin including MDP was shown to be effective in bonding to human enamel, and this effect was enhanced in combination with phosphate treatment.

Preparation of miniaturized hydrophilic affinity monoliths: Towards a reduction of non-specific interactions and an increased target protein density.

In affinity chromatography, non-specific interactions between the ligands and the affinity column may affect the results, leading to misinterpretations during the investigation of protein-ligand interactions (detection of false positives in ligand screening, lack of specificity in purification). Such non-specific interactions may arise both from the underlying support or from the target protein itself. If the second ones are protein-dependent (and cannot be studied in a general framework), the first ones occur in the same way regardless of the immobilized target. We propose a methodology to identify the origin of such non-specific interactions with the underlying material of the affinity column. This methodology relies on the systematic investigation of the retention behavior of a set of 41 low-molecular weight compounds covering a wide chemical space (net charge, log D, functionality). We first demonstrate that the main source of non-specific interactions on the most commonly used GMA-co-EDMA monolith comes from hydrophobic effects. To reduce such non-specific interactions, we developed a new hydrophilic glycidyl methacrylate-based monolith by replacing the EDMA crosslinker by the more hydrophilic NN' Methylenebisacrylamide (MBA). Optimization of the synthesis parameters (monomer content, initiation type, temperature) has focused on the reduction of non-specific interaction with the monolithic support while maximizing the amount of protein that can be grafted onto the monolith at the issue of its synthesis. The retention data of the 41 test solutes on the new poly(GMA-co-MBA) monolith shows a drastic reduction of non-specific interactions except for cationic compounds. The particular behavior of cationic compounds is due to their electrostatic interactions with carboxylic groups resulting from the partial acidic hydrolysis of amide groups of MBA during the epoxide ring opening step. So, the ring opening step in acidic media was replaced by a hot water treatment to avoid side reaction on MBA. The new monolith poly(GMA-co-MBA) not only has improved hydrophilic surface properties but also a higher protein density (16 ± 0.8 pmol cm-1 instead of 8 ± 0.3 pmol cm-1). To highlight the benefits of this new hydrophilic monolith for affinity chromatographic studies, frontal affinity chromatography experiments were conducted on these monoliths grafted with con A.

Influence of Test Specimen Geometry and Water Soaking on the In Vitro Cytotoxicity of Orthocryl®, Orthocryl® LC, Loctite® EA 9483 and Polypropylene.

Depending on their composition, plastics have a cytotoxic potential that needs to be evaluated before they are used in dentistry, e.g., as orthodontic removable appliances. Relevant guidelines set out requirements that a potential new resin in the medical field must meet, with a wide scope for experimental design. In the present study, test specimens of different geometries consisting of varying polymers (Orthocryl®, Orthocryl® LC, Loctite® EA 9483, Polypropylene) were soaked for different periods of time, then transferred to cell culture medium for 24 h, which was subsequently used for 24-h cultivation of A549 cells, followed by cytotoxicity assays (WST-1, Annexin V-FITC-propidium iodide (PI) flow cytometry). In this context, a reduction in the cytotoxic effect of the eluates of test specimens prepared from Orthocryl® LC and Loctite® EA 9483 was particularly evident in the Annexin V-FITC-PI assay when the soaking time was extended to 48 h and 168 h, respectively. Consistent with this, a reduced release of potentially toxic monomers into the cell culture medium, as measured by gas chromatography-mass spectrometry, was observed when the prior soaking time of test specimens of all geometries was extended. Remarkably, a significant increase in cytotoxic effect was observed in the WST-1 assay, which was accompanied by a higher release of monomers when the thickness of the test sample was increased from 0.5 to 1.0 mm, although an elution volume adapted to the surface area was used. However, further increasing the thickness to 3.0 mm did not lead to an increase in the observed cytotoxicity or monomer release. Test specimens made of polypropylene showed no toxicity under all test specimen sizes and soaking time conditions. Overall, it is recommended to perform toxicity studies of test specimens using different geometries and soaking times. Thereby, the influence of the different specimen thicknesses should also be considered. Finally, an extension of the test protocols proposed in ISO 10993-5:2009 should be considered, e.g., by flow cytometry or monomer analysis as well as fixed soaking times.

Effects of mussel-inspired co-deposition of 2-hydroxymethyl methacrylate and poly (2-methoxyethyl acrylate) on the hydrophilicity and binding tendency of common hemodialysis membranes: Molecular dynamics simulations and molecular docking studies.

Despite advances in the field, hemoincompatibility remains a critical issue for hemodialysis (HD) as interactions between various human blood constituents and the polymeric structure of HD membranes results in complications such as activation of immune system cascades. Adding hydrophilic polymer structures to the membranes is one modification approach that can decrease the extent of protein adsorption. This study conducted molecular dynamics (MD) simulations to understand the interactions between three human serum proteins (fibrinogen [FB], human serum albumin, and transferrin) and common HD membranes in untreated and modified forms. Poly(aryl ether sulfone) (PAES) and cellulose triacetate were used as the common dialyzer polymers, and membrane modifications were performed with 2-hydroxymethyl methacrylate (HEMA) and poly (2-methoxyethyl acrylate) (PMEA), using polydopamine-assisted co-deposition. The MD simulations were used as the framework for binding energy simulations, and molecular docking simulations were also performed to conduct molecular-level investigations between the two modifying polymers (HEMA and PMEA) and FB. Each of the three proteins acted differently with the membranes due to their unique nature and surface chemistry. The simulations show PMEA binds less intensively to FB with a higher number of hydrogen bonds, which reflects PMEA's superior performance compared to HEMA. The simulations suggest PAES membranes could be used in modified forms for blood-contact applications as they reflect the lowest binding energy to blood proteins.

Bioinspired Durable Antibacterial and Antifouling Coatings Based on Borneol Fluorinated Polymers: Demonstrating Direct Evidence of Antiadhesion.

Substituting natural products for traditional poison-killing antifouling agents is an efficient and promising method to alleviate the increasingly serious ecological crisis and aggravate the loss due to marine biofouling. Herein, the successful synthesis of poly(methyl methacrylate-co-ethyl acrylate-co-hexafluorobutyl methacrylate-co-isobornyl methacrylate) copolymer (PBAF) with borneol monomers and fluorine by a free radical polymerization method is reported. The PBA0.09F coating exhibits outstanding antibacterial and antifouling activity, achieving 98.2% and 92.3% resistance to Escherichia coli and Staphylococcus aureus, respectively, and the number of Halamphora sp. adhesion is only 26 (0.1645 mm2) in 24 h. This remarkable antibacterial and antifouling performance is attributed to the incorporation of fluorine components into the copolymer, which induces a low surface energy and hydrophobicity and the complex molecular structure of the natural nontoxic antifouling agent borneol. In addition, the results showed that the contents of the adhesion-related proteins mfp-3, mfp-5, and mfp-6 were significantly reduced, which proved that natural substances affect the secretion of biological proteins. Importantly, the PBAF coating exhibits excellent environmental friendliness and long-term stability. The antifouling mechanism is clarified, and an effective guide for an environmentally friendly antifouling coating design is proposed.

Surface Plasmon Resonance Based on Molecularly Imprinted Polymeric Film for l-Phenylalanine Detection.

In this study, we designed a simple, rapid, sensitive and selective surface plasmon resonance (SPR) sensor for detection of L-phenylalaine by utilizing molecular imprinting technology. l-phenylalanine imprinted and non-imprinted poly(2-hydroxyethyl methacrylate-methacryloyl-l-phenylalanine) polymeric films were synthesized onto SPR chip surfaces using ultraviolet polymerization. l-phenyalanine imprinted and non-imprinted SPR sensors were characterized by using contact angle, atomic force microscopy and ellipsometry. After characterization studies, kinetic studies were carried out in the concentration range of 5.0-400.0 µM. The limit of detection and quantification were obtained as 0.0085 and 0.0285 µM, respectively. The response time for the test including equilibration, adsorption and desorption was approximately 9 min. The selectivity studies of the l-phenylalanine imprinted SPR sensor was performed in the presence of d-phenylalanine and l-tryptophan. Validation studies were carried out via enzyme-linked immunosorbent analysis technique in order to demonstrate the applicability and superiority of the l-phenylalanine imprinted SPR sensor.

El grafeno en la rehabilitación bucal y su comportamiento biológico: caso clínico / Graphene in oral rehabilitation and its biological behavior: a clinical case

El grafeno y sus derivados son muy utilizados en ciencia y tecnología por los beneficios que otorgan sus propiedades fisicoquímicas. En el área de la salud en particular, se destacan sus propiedades biológicas debido a su elevada biocompatibilidad, interacción celular y su actividad antibacteriana. La incorporación de grafeno en ciertos materiales permite obtener un material combinado con propiedades mejoradas. Un ejemplo de ello es la incorporación industrial de óxido de grafeno en metacrilato de metilo para generar un polímero (PMMA) mejorado, no solo desde el punto de vista mecánico, sino también una notoria ventaja en la respuesta biológica de los tejidos blandos. Este artículo describe el caso clínico de un paciente de 70 años, que concurrió a la consulta buscando alternativas de tratamiento para mejorar la retención y estabilidad de las prótesis para optimizar la función masticatoria, una alternativa que impacte positivamente sobre su calidad de vida. El plan de tratamiento contempló el reemplazo de las prótesis removibles por prótesis híbridas en ambos maxilares, confeccionadas con PMMA modificado industrialmente con óxido de grafeno, previa colocación de cinco implantes en cada arco (AU)

Graphene and its derivatives are widely used in science and technology due to the benefits provided by their physicochemical properties. In the health area, specifically, its biological properties stand out, due to its high biocompatibility, cellular interaction, and its antibacterial activity. The incorporation of graphene in certain materials allows obtaining a combined material with improved properties. An example of this is the industrial incorporation of graphene oxide in methyl methacrylate, to generate an improved polymer (PMMA), not only from a mechanical point of view, but also a notable advantage in the biological response of soft tissues. This article describes the clinical case of a 70-year-old patient, who attended the consultation looking for treatment alternatives to improve the retention and stability of the prostheses to optimize the masticatory function, or an alternative that had a positive impact on their quality of lifetime. The treatment plan contemplated the replacement of removable prostheses with hybrid prostheses in both jaws, made with PMMA industrially modified with graphene oxide, after placing five implants in each arch (AU)

Synthesis of Block Copolymer Brush by RAFT and Click Chemistry and Its Self-Assembly as a Thin Film.

A well-defined block copolymer brush poly(glycidyl methacrylate)-graft-(poly(methyl methacrylate)-block- poly(oligo(ethylene glycol) methyl ether methacrylate)) (PGMA-g-(PMMA-b-POEGMA)) is synthesized via grafting from an approach based on a combination of click chemistry and reversible addition-fragmentation chain transfer (RAFT) polymerization. The resulting block copolymer brushes were characterized by 1H-NMR and size exclusion chromatography (SEC). The self-assembly of the block copolymer brush was then investigated under selective solvent conditions in three systems: THF/water, THF/CH3OH, and DMSO/CHCl3. PGMA-g-(PMMA-b-POEGMA) was found to self-assemble into spherical micelle structures as analyzed by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The average size of the particles was much smaller in THF/CH3OH and DMSO/CHCl3 as compared with the THF/water system. Thin film of block copolymer brushes with tunable surface properties was then prepared by the spin-coating technique. The thickness of the thin film was confirmed by scanning electron microscopy (SEM). Atom force microscopy (AFM) analysis revealed a spherical morphology when the block copolymer brush was treated with poor solvents for the backbone and hydrophobic side chains. The contact angle measurements were used to confirm the surface rearrangements of the block copolymer brushes.

Mixed receptors of AMPA and NMDA emulated using a 'Polka Dot'-structured two-dimensional conjugated polymer-based artificial synapse.

In a biological synapse, α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors mediate fast excitatory neurotransmission, whereas N-methyl-d-aspartate (NMDA) receptors trigger an enhanced memory effect; the complementary roles of AMPA and NMDA are essential in short-term plasticity (STP) to enhance memory effect (EME) transition. Herein, we report the design and fabrication of the first two-dimensional (2D) conjugated polymer (CP)-based synaptic transistor. The special design of the 2D CP with nanoscale-segregated 'polka dot'-structured crystalline phases and adjacent amorphous phases emulate the different receptors of NMDA and AMPA on the postsynaptic membrane for the first time. The synergistic effect of mixed receptors distinguishes STP and enhanced memory effect with a critical point, which regulates the threshold level of the enhanced memory effect induction. This effect has not been reported yet. The special structure avoids easy saturation of a single receptor with consecutively increased excitatory postsynaptic current (EPSC) in response to 1200 stimuli. Furthermore, the 2D P3HT synapse successfully emulates activity-dependent synaptic plasticity, such as metaplasticity and homeostatic plasticity, which are advanced forms of plasticity, allowing the self-adaptive ability of a synapse, but have rarely been reported.

Novel Osteogenic Behaviors around Hydrophilic and Radical-Free 4-META/MMA-TBB: Implications of an Osseointegrating Bone Cement.

Poly(methyl methacrylate) (PMMA)-based bone cement, which is widely used to affix orthopedic metallic implants, is considered bio-tolerant but lacks osteoconductivity and is cytotoxic. Implant loosening and toxic complications are significant and recognized problems. Here we devised two strategies to improve PMMA-based bone cement: (1) adding 4-methacryloyloxylethyl trimellitate anhydride (4-META) to MMA monomer to render it hydrophilic; and (2) using tri-n-butyl borane (TBB) as a polymerization initiator instead of benzoyl peroxide (BPO) to reduce free radical production. Rat bone marrow-derived osteoblasts were cultured on PMMA-BPO, common bone cement ingredients, and 4-META/MMA-TBB, newly formulated ingredients. After 24 h of incubation, more cells survived on 4-META/MMA-TBB than on PMMA-BPO. The mineralized area was 20-times greater on 4-META/MMA-TBB than PMMA-BPO at the later culture stage and was accompanied by upregulated osteogenic gene expression. The strength of bone-to-cement integration in rat femurs was 4- and 7-times greater for 4-META/MMA-TBB than PMMA-BPO during early- and late-stage healing, respectively. MicroCT and histomorphometric analyses revealed contact osteogenesis exclusively around 4-META/MMA-TBB, with minimal soft tissue interposition. Hydrophilicity of 4-META/MMA-TBB was sustained for 24 h, particularly under wet conditions, whereas PMMA-BPO was hydrophobic immediately after mixing and was unaffected by time or condition. Electron spin resonance (ESR) spectroscopy revealed that the free radical production for 4-META/MMA-TBB was 1/10 to 1/20 that of PMMA-BPO within 24 h, and the substantial difference persisted for at least 10 days. The compromised ability of PMMA-BPO in recruiting cells was substantially alleviated by adding free radical-scavenging amino-acid N-acetyl cysteine (NAC) into the material, whereas adding NAC did not affect the ability of 4-META/MMA-TBB. These results suggest that 4-META/MMA-TBB shows significantly reduced cytotoxicity compared to PMMA-BPO and induces osteoconductivity due to uniquely created hydrophilic and radical-free interface. Further pre-clinical and clinical validations are warranted.

Well-defined hydrophilic "turn-on"-type ratiometric fluorescent molecularly imprinted polymer microspheres for direct and highly selective herbicide optosensing in the undiluted pure milks.

Molecularly imprinted polymer (MIP)-based optosensing materials capable of direct, reliable, and highly selective detection of small organic analytes in complex aqueous samples hold great promise in many bioanalytical applications, but their development remains a challenging task. Addressing this issue, well-defined hydrophilic "turn-on"-type ratiometric fluorescent MIP microspheres are developed via a versatile and modular strategy based on the controlled/"living" radical polymerization method. Its general principle was demonstrated by the synthesis of red CdTe quantum dot (QD)-labeled silica particles with surface-bound atom transfer radical polymerization (ATRP)-initiating groups via the one-pot sol-gel reaction and their successive grafting of a thin fluorescent 2,4-D (an organic herbicide)-MIP layer (labeled with green organic fluorophores bearing both nitrobenzoxadiazole (NBD) and urea interacting groups) and hydrophilic poly(glyceryl monomethacrylate) (PGMMA) brushes via surface-initiated ATRP. The introduction of PGMMA brushes and rationally selected dual fluorescence labeling (i.e., red CdTe QDs being inert to 2,4-D and green NBD showing fluorescence "light-up" upon binding 2,4-D) onto MIP particles afforded them excellent complex aqueous sample-compatibility (due to their largely enhanced hydrophilicity) and analyte binding-induced "turn-on"-type ratiometric fluorescence changes, respectively. Such advanced MIP particles proved to be promising optosensing materials, which had a detection limit of 0.13 µM and showed obvious fluorescent color change upon binding different concentrations of 2,4-D in the undiluted pure milk. Moreover, they were successfully applied for direct and highly selective quantification of 2,4-D in the undiluted pure goat and bovine milks with good recoveries (97.9%-104.5%), even in the presence of several analogues of 2,4-D.

Platinum Nanoparticles Obtained at Mild Conditions on S-Layer Protein/Polymer Particle Supports.

This work presents the synthesis of platinum nanoparticles supported on S-layer protein/polymeric particle systems, obtained by combining proteins isolated from Lactobacillus kefiri and an aqueous dispersion of acrylic particles. FTIR spectra of the protein/polymer supports did not show changes in the Amide I band of the proteins, suggesting that proteins maintained their conformation after adsorption. The SAXS spectra and DLS results are consistent with the formation of a protein corona around the polymer particles. After combining the supports with the platinum complex and subsequently reducing the combination with hydrogen at mild conditions, we obtained colloidal nanocomposite materials. In these, platinum nanoparticles with diameters around 3 nm located on the surface of the protein/polymer supports were observed by TEM. The obtained nanosystems showed catalytic activity in the reduction of p-nitrophenol with NaBH4 at room temperature with conversions of 100% for reaction times of 50 to 70 min.

One-Step Biotinylation of Cellulose Paper by Polymer Coating to Prepare a Paper-Based Analytical Device.

Cellulose paper has strong potential as an analytical platform owing to its unique characteristics. In the present study, we investigated a procedure for functionalizing the surface of cellulose paper by dip-coating a mixture of a functional polymer and a perfluoroalkylated surfactant (surfactant 1). The functional polymer comprised a mixture of methyl methacrylate and poly(ethylene glycol) methacrylate monomers. The monomer ratio in the functional polymer affected the hydrophilicity and water absorbance of the cellulose paper after dip-coating. Furthermore, the presence of surfactant 1 during dip-coating promoted the surface segregation of poly(ethylene glycol) (PEG) moieties in the polymer, which enhanced the hydrophilicity, prevented nonspecific protein adsorption, and maintained the water absorbance of the dip-coated cellulose paper. Dip-coating with another functional polymer containing biotin groups produced a cellulose paper with a biotin-decorated surface in a one-step procedure. The displayed biotin groups immobilized avidin on the surface, and the PEG moieties in the polymer prevented nonspecific protein adsorption. We then immobilized a thrombin-binding DNA aptamer on the avidin-immobilized cellulose paper to prepare a paper-based analytical device. It is possible to visualize thrombin in model solutions and serum using the paper-based analytical device.

Hybrid monoliths with metal-organic frameworks in spin columns for extraction of non-steroidal drugs prior to their quantitation by reversed-phase HPLC.

A (glycidyl methacrylate)-co-(ethylene glycol dimethacrylate) polymer (poly(GMA-co-EDMA)) was functionalized with metal-organic frameworks (MOF) and used as a sorbent for solid-phase extraction (SPE). The polymeric sorbent was prepared in-situ by photopolymerization in a previously wall-modified spin column, and then modified with an amino-modified MOF of type NH2-MIL-101(Cr). The sorbents were used for the extraction of nonsteroidal anti-inflammatory drugs (NSAIDs) from human urine samples. The sorbent was compared with the parent monolith and embedded approach, where the MOF particles are admixed in the polymerization mixture before the in-situ polymerization in the modified spin column. SPE is performed by percolating the sample solutions in a centrifuge, which streamlines the SPE steps. The hybrid composites were characterized by scanning electron microscopy and nitrogen intrusion porosimetry. Three NSAIDs (ketoprofen, flurbiprofen, and ibuprofen) were tested. They were eluted from the sorbent with acidified water-acetonitrile mixtures and subsequently analyzed by reversed-phase HPLC with UV detection. The detection limits varied in the range from 0.1 to 7 µg·L-1, and the precisions (relative standard deviation) were <14% in all the cases. The recoveries were between 71.0 and 78.0% in spiked urine samples. Graphical abstractA hybrid monolith modified with amino-modified MOF [named NH2-MIL-101(Cr)] in wall-modified spin columns was prepared. The resulting micro-extraction device was applied to the extraction and preconcentration of non-steroidal anti-inflammatory drugs.

Covalent organic framework incorporated chiral polymer monoliths for capillary electrochromatography.

A covalent organic framework, Schiff base network-1 (SNW-1), was synthesized and incorporated into cellulase based poly(glycidyl methacrylate-co-ethylene dimethacrylate) (cellulase@poly(GMA-EDMA-SNW-1)) monolith to afford a novel chiral stationary phase for capillary electrochromatography (CEC). SNW-1 is attractive as a stationary phase for CEC because it not only features high surface areas but also provides conjugate structures and abundant amine groups to give π-π electrostatic stacking and hydrogen bonding property. Incorporation of SNW-1 into monolithic column could improve the column efficiency and increase the interactions between the tested racemates and the stationary phase thus significantly improved their CEC separation. The obtained monoliths were characterized by scanning electron microscopy, elemental analysis and nitrogen adsorption. Moreover, effects of SNW-1 concentration, immobilization pH of cellulase and CEC conditions were also investigated. Under the optimized conditions, the cellulase@poly(GMA-EDMA-SNW-1) monolith exhibited excellent enantioseparation performance for eight pairs of different classes of chiral drugs including ß-blockers, antihistamines and anticoagulants. Satisfactory repeatability was achieved with relative standard deviations for intra-day, inter-day and column-to-column runs less than 4.5%, and batch-to-batch runs less than 6.8%. The experiment results reveal that the combination of the versatile features of monoliths and unique properties of SNW-1 could be a promising strategy for chiral separation.

Biodrug Suppresses Breast and Colorectal Cancer in Murine Models.

RNA interference (RNAi) remains one of the most promising and emerging strategies for the effective cancer treatment due to its high target specificity and greater potency. However, it is hindered due to lack of appropriate targeting technologies. Therefore, there is an imminent need to develop specific and robust delivery systems for successful gene silencing. Nanotechnology-based strategies have been in place to combat the shortcomings associated with viral-based delivery systems. Herein we describe protocols for successful in vitro and in vivo delivery of gene-specific nucleic acids such as siRNAs and shRNAs using PEI-PGMA nanoparticles for efficient cancer therapy.

Sensitivity of a continuous hot-melt extrusion and strand pelletization line to control actions and composition variation.

The purpose of this work was to develop a robust hot-melt extrusion and strand pelletization process for manufacturing pellets with an immediate release (IR) of a poorly water-soluble active pharmaceutical ingredient (API), nimodipine. The robustness of pharmaceutical continuous manufacturing processes and of its control strategy is vital for competitiveness to traditional batch-manufacturing. Therefore, first the sensitivity of product quality, process stability, and process monitoring tools to i) parameter changes due to control actions and ii) typical process deviations, i.e., feeding errors, was investigated in a design of experiments (DoE). Thereby, die melt pressure was found to be highly sensitive to composition deviations, i.e. a limiting factor for process stability. Especially critical were deviations to increased HPMC content, since it behaved as a filler in the melt. Pelletization, or pellet size and size distribution respectively, were found to be sensitive to an increased throughput, due to the resulting insufficient strand cooling before the pelletizer. API dissolution in contrast, was found to be robust across the entire investigated range of formulation and process settings. Second, a design space for the production of IR pellets for subsequent tableting was established, and finally, a technical control strategy was developed to ensure a robust process. Near-infrared (NIR) spectroscopy was applied to monitor API content and the sensitivity of the residence time distribution (RTD) was investigated by means of tracer measurements. NIR-based API content monitoring and RTD models for material tracking were found to be at risk after processing melt with high HPMC content, due to a lack of purging by less viscous formulation compositions.

Development of FAcE (Formulated Alhydrogel competitive ELISA) method for direct quantification of OAg present in Shigella sonnei GMMA-based vaccine and its optimization using Design of Experiments approach.

Many formulated vaccines, including 1790GAHB Shigella sonnei GMMA-based vaccine, contain Alhydrogel (aluminum hydroxide), consequently the antigen content must be determined in the formulated final vaccine product, as required by regulatory authorities. The direct quantification of antigens adsorbed on aluminum salts is difficult, and antigens may need to be extracted using laborious and often ineffective desorption procedures. To directly quantify the sugar vaccine target in the LPS of 1790GAHB, we have developed a new FAcE (Formulated Alhydrogel competitive ELISA) method. FAcE is an immunoassay based on the competition between S. sonnei LPS, coated on the ELISA plate, and the LPS in formulated S. sonnei GMMA, in binding a specific monoclonal antibody. To optimize the method, which is as easy to perform as a standard ELISA, we have applied a Design of Experiments (DOE) approach. A model was found to define the significant assay variables and to predict their impact on the output responses. Results obtained using the DOE optimized FAcE assay showed that the method is sensitive (0.02 µg/mL lower detection limit), precise, reproducible and can accurately quantify independently formulated drug products, making it a useful tool in routine tests of Alhydrogel-based vaccines. We are currently using this method to determine S. sonnei vaccine potency, stability and lot-to-lot variations, and are broadening its applicability to quantify active ingredients of other Alhydrogel GMMA-vaccines and in multivalent vaccines formulations.