Selection, characterization, and biosensing applications of DNA aptamers targeting cyanotoxin BMAA.

Scientists have established a connection between environmental exposure to toxins like ß-N-methylamino-l-alanine (BMAA) and a heightened risk of neurodegenerative disorders. BMAA is a byproduct from certain strains of cyanobacteria that are present in ecosystems worldwide and is renowned for its bioaccumulation and biomagnification in seafood. The sensitivity, selectivity, and reproducibility of the current analytical techniques are insufficient to support efforts regarding food safety and environment monitoring adequately. This work outlines the in vitro selection of BMAA-specific DNA aptamers via the systematic evolution of ligands through exponential enrichment (SELEX). Screening and characterization of the full-length aptamers was achieved using the SYBR Green (SG) fluorescence displacement assay. Aptamers BMAA_159 and BMAA_165 showed the highest binding affinities, with dissociation constants (Kd) of 2.2 ± 0.1 µM and 0.32 ± 0.02 µM, respectively. After truncation, the binding affinity was confirmed using a BMAA-conjugated fluorescence assay. The Kd values for BMAA_159_min and BMAA_165_min were 6 ± 1 µM and 0.63 ± 0.02 µM, respectively. Alterations in the amino proton region studied using solution nuclear magnetic resonance (NMR) provided further evidence of aptamer-target binding. Additionally, circular dichroism (CD) spectroscopy revealed that BMAA_165_min forms hybrid G-quadruplex (G4) structures. Finally, BMAA_165_min was used in the development of an electrochemical aptamer-based (EAB) sensor that accomplished sensitive and selective detection of BMAA with a limit of detection (LOD) of 1.13 ± 0.02 pM.

Bioactive Scaffolds as a Promising Alternative for Enhancing Critical-Size Bone Defect Regeneration in the Craniomaxillofacial Region.

Reconstruction of critical-size bone defects (CSDs) in the craniomaxillofacial (CMF) region remains challenging. Scaffold-based bone-engineered constructs have been proposed as an alternative to the classical treatments made with autografts and allografts. Scaffolds, a key component of engineered constructs, have been traditionally viewed as biologically passive temporary replacements of deficient bone lacking intrinsic cues to promote osteogenesis. Nowadays, scaffolds are functionalized, giving rise to bioactive scaffolds promoting bone regeneration more effectively than conventional counterparts. This review focuses on the three approaches most used to bioactivate scaffolds: (1) conferring microarchitectural designs or surface nanotopography; (2) loading bioactive molecules; and (3) seeding stem cells on scaffolds, providing relevant examples of in vivo (preclinical and clinical) studies where these methods are employed to enhance CSDs healing in the CMF region. From these, adding bioactive molecules (specifically bone morphogenetic proteins or BMPs) to scaffolds has been the most explored to bioactivate scaffolds. Nevertheless, the downsides of grafting BMP-loaded scaffolds in patients have limited its successful translation into clinics. Despite these drawbacks, scaffolds containing safer, cheaper, and more effective bioactive molecules, combined with stem cells and topographical cues, remain a promising alternative for clinical use to treat CSDs in the CMF complex replacing autografts and allografts.

Autonomous electrochemical system for ammonia oxidation reaction measurements at the International Space Station.

An autonomous electrochemical system prototype for ammonia oxidation reaction (AOR) measurements was efficiently done inside a 4'' x 4'' x 8'' 2U Nanoracks module at the International Space Station (ISS). This device, the Ammonia Electrooxidation Lab at the ISS (AELISS), included an autonomous electrochemical system that complied with NASA ISS nondisclosure agreements, power, safety, security, size constrain, and material compatibility established for space missions. The integrated autonomous electrochemical system was tested on-ground and deployed to the International Space Station as a "proof-of-concept" ammonia oxidation reaction testing space device. Here are discussed the results of cyclic voltammetry and chronoamperometry measurements done at the ISS with a commercially available channel flow-cell with eight screen-printed electrodes, including Ag quasi-reference (Ag QRE) and carbon counter electrodes. Pt nanocubes in Carbon Vulcan XC-72R were used as the catalyst for the AOR and 2 µL drop of Pt nanocubes/ Carbon Vulcan XC-72R, 20 wt%, ink was placed on the carbon working electrodes and allowed to dry in air. After the AELISS was prepared for launch to the ISS, a 4 days delayed (2 days in the space vehicle Antares and 2 days space transit to the ISS) cause a slight shift on the Ag QRE potential. Nevertheless, the AOR cyclic voltametric peak was observed in the ISS and showed ca. 70% current density decrease due to the buoyancy effect in agreement with previous microgravity experiments done at the zero-g aircraft.

Investigating the role of Prefrontal Neurons in cognitive tasks through Fiber Photometry and Adapted Barnes Maze Protocols

The medial prefrontal cortex (mPFC) is essential in the execution of cognitive tasks, however very little is known on how these neurons are modulated during specific tasks and which subtype of neurons are responsible for so. Therego, with the intention of addressing this issue, we recorded mPFC gabaergic and glutamatergic activation patterns through fiber photometry (FIP) in mice, while simultaneously performing the Barnes Maze (BM) cognitive task (4 day behavioral trial). In addition, an altered structural and procedural protocol for BM was validated in this study due to necessary modifications allowing FIP and BM to happen simultaneously. A successful protocol validation was followed by our preliminary results, which showed that both glutamatergic and gabaergic neurons presented significant change in activation intensity and number of events in specific contexts throughout the task days. In addition, when stratified and crossed with BM performance parameters, such as latency to complete tasks and adopted strategy, glutamatergic and gabaergic neurons presented a significant decline in both activation patterns and number of activation events throughout the days. This data suggest not only an important role of glutamatergic and gabaergic mPFC neurons in learning, memory and decision making, but also that activation patterns of each of these groups may serve as markers for cognitive progression and/or dysfunction. KEY-WORDS: Memory, Learning, Decision Making, Medial Prefrontal Cortex (mPFC), Fiber Photometry (FIP), Barnes Maze (BM), Glutamatergic, Gabaergic, Neuronal Activity, Neuronal Activation Patterns, Neuronal Dynamics.

O córtex pré-frontal medial (mPFC) é essencial na execução de tarefas cognitivas, no entanto, pouco se sabe sobre como esses neurônios são modulados durante tarefas específicas e qual subtipo de neurônios é responsável por isso. Portanto, com a intenção de abordar essa questão, registramos os padrões de ativação de neurônios gabaérgicos e glutamatérgicos do mPFC por meio de fotometria de fibra (FIP) em camundongos, enquanto realizávamos simultaneamente a tarefa cognitiva do Labirinto de Barnes (BM) (ensaio comportamental de 4 dias). Além disso, um protocolo estrutural e procedimental alterado para o BM foi validado neste estudo devido a modificações necessárias que permitiram a realização simultânea de FIP e BM. Uma validação bem-sucedida do protocolo foi seguida pelos nossos resultados preliminares, que mostraram que tanto os neurônios glutamatérgicos quanto os gabaérgicos apresentaram mudanças significativas na intensidade de ativação e no número de eventos em contextos específicos ao longo dos dias da tarefa. Além disso, quando estratificados e cruzados com parâmetros de desempenho do BM, como latência para completar as tarefas e estratégia adotada, os neurônios glutamatérgicos e gabaérgicos apresentaram uma diminuição significativa nos padrões de ativação e no número de eventos de ativação ao longo dos dias. Esses dados sugerem não apenas um papel importante dos neurônios glutamatérgicos e gabaérgicos do mPFC na aprendizagem, memória e tomada de decisões, mas também que os padrões de ativação de cada um desses grupos podem servir como marcadores de progressão e/ou disfunção cognitiva. PALAVRAS-CHAVE: Memória, Aprendizagem, Tomada de Decisões, Córtex Pré-Frontal Medial (mPFC), Fotometria de Fibra (FIP), Labirinto de Barnes (BM), Glutamatérgico, Gabaérgico, Atividade Neuronal, Padrões de Ativação Neuronal, Dinâmica Neuronal.

Transiently Nav1.8-expressing neurons are capable of sensing noxious stimuli in the brain.

While current research highlights the role of Nav1. 8 sensory neurons from the peripheral nervous system, the anatomical and physiological characterization of encephalic Nav1.8 neurons remains unknown. Here, we use a Cre/fluorescent reporter mouse driven by the Nav1.8 gene promoter to reveal unexpected subpopulations of transiently-expressing Nav1.8 neurons within the limbic circuitry, a key mediator of the emotional component of pain. We observed that Nav1.8 neurons from the bed nuclei of the stria terminalis (BST), amygdala, and the periaqueductal gray (vPAG) are sensitive to noxious stimuli from an experimental model of chronic inflammatory pain. These findings identify a novel role for central Nav1.8 neurons in sensing nociception, which could be researched as a new approach to treating pain disorders.

Bioactive Cellulose Acetate Electrospun Mats as Scaffolds for Bone Tissue Regeneration.

In the last decades, cell-based approaches for bone tissue engineering (BTE) have relied on using models that cannot replicate the complexity of the bone microenvironment. There is an ongoing amount of research on scaffold development responding to the need for feasible materials that can mimic the bone extracellular matrix (ECM) and aid bone tissue regeneration (BTR). In this work, a porous cellulose acetate (CA) fiber mat was developed using the electrospinning technique and the mats were chemically modified to bioactivate their surface and promote osteoconduction and osteoinduction. The mats were characterized using FTIR and SEM/EDS to validate the chemical modifications and assess their structural integrity. By coupling adhesive peptides KRSR, RGD, and growth factor BMP-2, the fiber mats were bioactivated, and their induced biological responses were evaluated by employing immunocytochemical (ICC) techniques to study the adhesion, proliferation, and differentiation of premature osteoblast cells (hFOB 1.19). The biological assessment revealed that at short culturing periods of 48 hours and 7 days, the presence of the peptides was significant for proliferation and adhesion, whereas at longer culture times of 14 days, it had no significant effect on differentiation and maturation of the osteogenic progenitor cells. Based on the obtained results, it is thus concluded that the CA porous fiber mats provide a promising surface morphology that is both biocompatible and can be rendered bioactive upon the addition of osteogenic peptides to favor osteoconduction leading to new tissue formation.

Porous Cellulose Acetate/Block Copolymer Membranes for the Recovery of Polyphenolic Compounds from Aquatic Environments.

Polyphenols are natural compounds with strong antioxidant properties synthesized by plants and widely distributed in plant tissues. They compose a broad class of compounds that are commonly employed for multiple applications such as food, pharmaceutical, adhesives, biomedical, agricultural, and industrial purposes. Runoffs from these sources result in the introduction of polyphenols into aquatic environments where they further transform into highly toxic pollutants that can negatively affect aquatic ecosystems and humans. Therefore, the development of extraction and remediation methods for such compounds must be addressed. This study describes the identification and operation of a method to recover polyphenolic compounds from water environments by utilizing membrane-based separation. Composite membranes derived from electrospun cellulose acetate (CA) fibers and diblock copolymer (DiBCP) PEO-b-P4VP were prepared to evaluate the adsorption of polyphenolic compounds from aqueous environments. The highly porous CA fibers were developed using the electrospinning technique, and the fabricated DiBCP/CA membranes were characterized using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), Fourier-transform infrared (FT-IR) spectroscopy, and tensile testing. Finally, the ability of the composite membranes to adsorb the soluble polyphenolic compounds catechol (CAT) and gallic acid (GA), from a wetland environment, was studied via batch adsorption experiments and by solid-phase extraction (SPE). Results revealed a successful recovery of both polyphenols, at concentrations within the parts per million (ppm) range, from the aqueous media. This suggests a novel approach to recover these compounds to prevent their transformation into toxic pollutants upon entrance to water environments.

The influence of calcium ions (Ca2+) on the enzymatic hydrolysis of lipopolysaccharide aggregates to liberate free fatty acids (FFA) in aqueous solution.

The chemical environment in aqueous solutions greatly influences the ability of amphiphilic molecules such as lipopolysaccharides (LPS) to aggregate into different structural phases in aqueous solutions. Understanding the substrate's morphology and conditions of aqueous solution that favor both enzymatic activity and the disruption of LPS aggregates are crucial in developing agents that can counteract the new trend of multidrug resistance by gram-negative bacteria. In this study, we developed two LPS morphologies using LPS from Escherichia coli as a model to study the in vitro hydrolytic response when using a lipase treatment. The hydrolysis was performed using lipase b from Candida antarctica to understand the catalytic effect in removing fatty acids from its lipid A moiety on different LPS aggregates. Physical and chemical characterizations of the products included dynamic light scattering, small angle X-ray scattering, Fourier transform infrared spectroscopy, thin-layer chromatography, and gas chromatography. Our results suggest a trend of prominent hydrolytic response (72% enhancement) upon the addition of calcium ions to induce LPS aggregates into bilayer formations. Moreover, our results revealed the detection of myristic acid (C14:0) as the product of the hydrolysis when using RaLPS in its aggregate forms.

Genetic association of the PERIOD3 (PER3) Clock gene with extreme obesity.

BACKGROUND: Obesity has reached epidemic proportions worldwide, affecting life quality and span. Susceptibility to obesity is partly mediated by genetic differences. Indeed, several genes from the clock gene family have already been shown to be intimately associated with obesity in diverse ethnic groups. In the present study, an association between BMI and the rs707467, rs228697 and rs228729 PER3 (Period Circadian Clock 3) polymorphisms in subjects with class II (BMI ≥ 35.0-39.9 kg/m2) and class III obesity (>40 kg/m2, extreme obesity) were carried out using TaqMan real-time PCR. Overall, 259 Brazilian adults were genotyped, of whom 122 had class II or III obesity (BMI ≥ 35.0 kg/m2) and 137 were controls having normal weight (BMI > 18.5 and <24.9 kg/m2). RESULTS: PER3 tag SNP (rs228729) shows a significant association with extreme obesity (1000 permutation p = 0.03 and p = 0.04), for genotype and allele frequency respectively) and a haplotype among the three assessed SNPs (alleles G/T/A, rs228697, rs228729, and rs707467, respectively, 1000 permutation p = 0.03) was significantly more prevalent in the group with obesity. CONCLUSION: This exploratory association study suggests that PER3 rs228729 may be associated with extreme obesity in Brazilian adults, however, replication is needed.

Performance Evaluation of 1-Cyclohexylpiperidine as a Draw Solute for Forward Osmosis Water Separation and CO2 Recovery.

Membrane-based technologies, such as forward osmosis (FO), offer the advantage of treating water through a spontaneous process that requires minimal energy input while achieving favorable water permeability and selectivity. However, the FO process still has some challenges that need to be solved or improved to become entirely feasible. The main impediment for this technology is the recovery of the draw solute used to generate the osmotic potential in the process. In this paper, we discuss the use of a switchable polarity solvent, 1-cyclohexylpiperidine (CHP), as a draw solute that responds to external stimuli. Specifically, the miscibility of CHP can be switched by the presence of carbon dioxide (CO2) and is reversible by applying heat. Thus, in this study, the hydrophobic CHP is first converted to the hydrophilic ammonium salt (CHPH+), and its capability as a draw solution (DS) is thoroughly evaluated against the typical osmotic agent, sodium chloride (NaCl). Our results show that the water permeability across the thin film composite membrane increases by 69% when CHPH+ is used as the DS. Also, the water permeability when using different feed solutions: aqueous solutions of (a) urea and (b) NaCl were evaluated. In both cases, the CHPH+ generates water fluxes in the range of 65 ± 4 LMH and 69 ± 2 LMH, respectively. We then separate the diluted DS by applying 75 °C to the solution to recover the pure CHP and water. The results of this work provide a proof-of-concept of a CHP wastewater and desalination method via an FO process.

Antimicrobial Polymer-Peptide Conjugates Based on Maximin H5 and PEG to Prevent Biofouling of E. coli and P. aeruginosa.

Many pathogens, such as Pseudomonas aeruginosa and Escherichia coli bacteria can easily attach to surfaces and form stable biofilms. The formation of such biofilms in surfaces presents a problem in environmental, biomedical, and industrial processes, among many others. Aiming to provide a plausible solution to this issue, the anionic and hydrophobic peptide Maximin H5 C-terminally deaminated isoform (MH5C) has been modified with a cysteine in the C-terminal (MH5C-Cys) and coupled to polyethylene glycol (PEG) polymers of varying sizes (i.e., 2 kDa and 5 kDa) to serve as a surface protective coating. Briefly, the MH5C-Cys was bioconjugated to PEG and purified by size exclusion chromatography while the reaction was confirmed via SDS-PAGE and MALDI ToF. Moreover, the preventive antimicrobial activity of the MH5C-Cys-PEG conjugates was performed via the growth curves method, showing inhibition of bacterial growth after 24 h. The efficacy of these peptide-polymer conjugates was extensively characterized via scanning electron microscopy (SEM), minimum inhibition concentration (MIC), minimum biofilm inhibition concentration (MBIC), and minimum biofilm eradication concentration (MBEC) assays to evaluate their ability to eradicate and prevent the biofilms. Interestingly, this work demonstrated a critical PEG polymer weight of 5 kDa as ideal when coupled to the peptide to achieve inhibition and eradication of the biofilm formation in both bacteria strains. According to the MICs (40 µM) and MBICs (300 µM), we can conclude that this conjugate (MH5C-Cys-5 kDa) has an action that prevents/inhibits the formation of biofilms and the eradication of biofilms (MBEC 500 µM). In contrast, the MH5C-Cys peptide with PEG polymer of 2 kDa did not show inhibition or eradication of the biofilms.

Lyotropic Liquid Crystal-Based Membranes for Water Remediation: Fabrication, Characterization and Performance Evaluation.

In water remediation, biomimetic membranes are gaining much attention due to their selectivity, dynamic stability, nontoxicity, and biocompatibility. Lyotropic liquid crystals (LLCs) are self-organizing networks that can conform to an array of geometries with high pore densities. As such, LLCs are excellent membrane materials for water applications because they are water insoluble and are manipulated to conform to an array of morphologies that provide natural water channels that are readily tunable in size. They have the ability to create uniform pores, between the range of 1 and 5 nm, with large surface areas. Thus, this work focuses on the design, fabrication, and characterization of LLC-modified Janus-type membranes for forward osmosis applications. Physical characterization of the membranes was performed using scanning electron microscopy (SEM), and the results show an open-pore radius and the presence of both finger- and sponge-like pores depending on membrane preparation. The contact angle assessment indicates that as the membranes are further modified with other polymers (e.g., PAN), higher hydrophilicity and surface energy are achieved. Moreover, the Brunauer-Emmett-Teller (BET) analysis showed a significant variation in the pore distribution between membranes. Functionalized membranes presented satisfactory water flux and superior salt rejection compared to nonfunctionalized membranes. SupPACMoDS membranes are 83% more efficient at preventing salt back flux than the nonmodified version. This is credited to the thickness and pore structure provided by the PAN support layer in the membrane.

Contribution of neuronal calcium sensor 1 (Ncs-1) to anxiolytic-like and social behavior mediated by valproate and Gsk3 inhibition.

Peripheral biomarker and post-mortem brains studies have shown alterations of neuronal calcium sensor 1 (Ncs-1) expression in people with bipolar disorder or schizophrenia. However, its engagement by psychiatric medications and potential contribution to behavioral regulation remains elusive. We investigated the effect on Ncs-1 expression of valproic acid (VPA), a mood stabilizer used for the management of bipolar disorder. Treatment with VPA induced Ncs-1 gene expression in cell line while chronic administration of this drug to mice increased both Ncs-1 protein and mRNA levels in the mouse frontal cortex. Inhibition of histone deacetylases (HDACs), a known biochemical effect of VPA, did not alter the expression of Ncs-1. In contrast, pharmacological inhibition or genetic downregulation of glycogen synthase kinase 3ß (Gsk3ß) increased Ncs-1 expression, whereas overexpression of a constitutively active Gsk3ß had the opposite effect. Moreover, adeno-associated virus-mediated Ncs-1 overexpression in mouse frontal cortex caused responses similar to those elicited by VPA or lithium in tests evaluating social and mood-related behaviors. These findings indicate that VPA increases frontal cortex Ncs-1 gene expression as a result of Gsk3 inhibition. Furthermore, behavioral changes induced by Ncs-1 overexpression support a contribution of this mechanism in the regulation of behavior by VPA and potentially other psychoactive medications inhibiting Gsk3 activity.

Characterization of non-covalent immobilized Candida antartica lipase b over PS-b-P4VP as a model bio-reactive porous interface.

The design of interfaces that selectively react with molecules to transform them into compounds of industrial interest is an emerging area of research. An example of such reactions is the hydrolytic conversion of ester-based molecules to lipids and alcohols, which is of interest to the food, and pharmaceutical industries. In this study, a functional bio-interfaced layer was designed to hydrolyze 4-nitrophenyl acetate (pNPA) and Ricinus Communis (castor) oil rich in triglycerides using lipase b from Candida antarctica (CALB, EC 3.1.1.3). The attachment of CALB was performed via non-covalent immobilization over a polymer film of vertically aligned cylinders that resulted from the self-assembly of the di-block copolymer polystyrene-block-poly(4-vinyl pyridine) (PS-b-P4VP). This polymer-lipase model will serve as the groundwork for the design of further bioactive layers for separation applications requiring similar hydrolytic processes. Results from the fabricated functional bio-interfaced material include cylinders with featured pore size of 19 nm, d spacing of 34 nm, and ca. 40 nm of thickness. The polymer-enzyme layers were physically characterized using AFM, XPS, and FTIR. The immobilized enzyme was able to retain 91% of the initial enzymatic activity when using 4-nitrophenyl acetate (pNPA) and 78% when exposed to triglycerides from castor oil.

Brain-enriched MicroRNA-184 is downregulated in older adults with major depressive disorder: A translational study.

Changes in microRNAs (miRNAs) expression have been described in major depressive disorder in young and middle-aged adults. However, no study has evaluated miRNA expression in older adults with major depression (or late-life depression [LLD]). Our primary aim was to evaluate the expression of miRNAs in subjects with LLD. We first evaluated the miRNA expression using next-generation sequencing (NGS) and then we validated the miRNAs found in NGS in an independent sample of LLD patients, using RT-qPCR. Drosophila melanogaster model was used to evaluate the impact of changes in miRNA expression on behavior. NGS analysis showed that hsa-miR-184 (log2foldchange = -4.21, p = 1.2 × 10-03) and hsa-miR-1-3p (log2foldchange = -3.45, p = 1.3 × 10-02) were significantly downregulated in LLD compared to the control group. RT-qPCR validated the downregulation of hsa-miR-184 (p < 0.001), but not for the hsa-miR-1-3p. The knockout flies of the ortholog of hsa-miR-184 showed significantly reduced locomotor activity at 21-24 d.p.e (p = 0.04) and worse memory retention at 21-24 d.p.e (24h post-stimulus, p = 0.02) compared to control flies. Our results demonstrated that subjects with LLD have significant downregulation of hsa-miR-184. Moreover, the knockout of hsa-miR-184 in flies lead to depressive-like behaviors, being more pronounce in older flies.

Nanocellulose-Block Copolymer Films for the Removal of Emerging Organic Contaminants from Aqueous Solutions.

The prevalence of emerging organic contaminants (EOCs) in ground and surface water has sparked the search for more effective methods to remove EOCs from the environment. In pursuit of a solution for this environmental concern, herein we present the development of reusable films based on cellulose nanofibers (CNFs) and the block copolymer, poly(4-vinylpyridine-b-ethylene oxide) (P4VP-PEO) to adsorb sulfamethoxazole (SMX) as an EOC model compound. We hypothesize that the adsorption of SMX was achieved mainly by π-π interactions between the pyridine functionalities of the block copolymer and the electron deficient phenyl group of the SMX. Preceding preparation of the films, CNFs were modified with the alkoxysilane trimethoxy(2-phenylethyl)silane (TMPES) to increase their stability in aqueous solution. After the addition of P4VP-PEO, the process was completed by filtration followed by oven-drying. XPS and FTIR were employed to confirm the addition of TMPES and P4VP-PEO, respectively. Adsorption batch experiments were performed in aqueous solutions of SMX at a neutral pH, obtaining adsorptions of up to 0.014 mmol/g in a moderate time of 60 min. For the reusability tests, films were immersed in ethanol 95 wt.% to elude the adsorbed SMX, rinsed with deionized (DI) water, and dried at room temperature to be reused in a new adsorption cycle. We found that this new composite material could be reused several times with negligible loss of adsorption capacity. The films presented have been shown to be of substantial importance for water remediation as they find direct application in the adsorption of electron deficient aromatic compounds and are reusable.

Fabrication and Evaluation of Polycaprolactone Beads-on-String Membranes for Applications in Bone Tissue Regeneration.

Tissue engineering leads to the development of biomaterial scaffolds where its biocompatibility and bioactivity are often improved after performing physical or chemical surface modification treatments. Micropatterning, soft lithography, and biofabrication are also approaches that provide a biomimetic microenvironment but have proven very costly and time consuming. In this concern, an appropriate substrate with suitable sites for cell attachment represents a major factor in cell behavior and biological functions. For this reason, our strategy was to fabricate a standard fibrous biomaterial with reproducible surface topography, incorporating microbeads and nanofeatures, and show the positive outcomes of the new substrate reflected on cell functions of bone cells. The electrospun polycaprolactone (PCL) beads-on-string membranes were obtained by adjusting the spinning solution at different concentrations until continuous beads were formed. Cell adhesion and proliferation, on the PCL scaffold, were analyzed the subsequent 2 days after initial culture. Complementary studies of cytoskeleton spreading and differentiation were analyzed after 7 and 14 days of the initial incubation. The scanning electron microscopy (SEM) images showed evidence of the formation of beads-on-string nanofibers and suggested that as-formed microstructures worked as attachment sites for osteoblasts. We investigated cell proliferation using anti-BrdU fluorescence assay, and results show a similar proliferation rate of cells cultured between PCL scaffolds and control. Finally, Phalloidin TRITC and antisialoprotein antibody were used to analyze cell spreading and differentiation after 7 and 14 days, respectively. This work shows a low-cost fabrication method to produce a biodegradable scaffold with micro/nanostructured characteristics that favor cell adhesion, proliferation, maturation, and subsequent differentiation of osteoblasts. According to the results, the biocompatibility of PCL beads-on-string could be comparable to other complex biomaterials, and we conclude that our scaffold is optimal for applications in bone tissue regeneration.

Cellulose Acetate/P4VP-b-PEO Membranes for the Adsorption of Electron-Deficient Pharmaceutical Compounds.

The prevalence of pharmaceutical compounds in surface and groundwater presents a rising threat to human health. As such, the search for novel materials that serve to avoid their release into the environment or for the remediation once in the water effluent is of utmost importance. The present work describes the fabrication of a cellulose acetate membrane modified with the block copolymer poly(4-vinylpyridine-b-ethylene oxide) (P4VP-b-PEO) crafted for the specific targeting and adsorption of electron-deficient pharmaceuticals (EDPs). The EDPs under study were sulfamethoxazole, sulfadiazine, and omeprazole. The results as part of this work present a thorough characterization of the prepared membranes by FTIR, contact angle measurement, and SEM images. Moreover, results show that the adsorptive character of the membrane correlates with the relative electron deficiency and spatial orientation of the contaminant. Interestingly, the addition of nominal 1% P4VP-b-PEO to the cellulose matrix helps to increase the adsorption efficiency of the membranes by at least 2-fold in most cases. For the compounds studied, the prepared membrane has a higher efficiency toward omeprazole followed by sulfamethoxazole and sulfadiazine. This work may serve to inspire the design and fabrication of selective soft materials for the adsorption and remediation of contaminants of emerging concern.