New Photocrosslinked 3D Foamed Scaffolds Based on GelMA Copolymers: Potential Application in Bone Tissue Engineering.

The production of customized polymeric hydrogels in the form of 3D scaffolds with application in bone tissue engineering is currently a topic of great interest. Based on gelatin methacryloyl (GelMa) as one of the most popular used biomaterials, GelMa with two different methacryloylation degrees (DM) was obtained, to achieve crosslinked polymer networks by photoinitiated radical polymerization. In this work, we present the obtention of new 3D foamed scaffolds based on ternary copolymers of GelMa with vinylpyrrolidone (VP) and 2-hydroxyethylmethacrylate (HEMA). All biopolymers obtained in this work were characterized by infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA), whose results confirm the presence of all copolymers in the crosslinked biomaterial. In addition, scanning electron microscopy (SEM) pictures were obtained verifying the presence of the porosity created by freeze-drying process. In addition, the variation in its swelling degree and its enzymatic degradation in vitro was analyzed as a function of the different copolymers obtained. This has allowed us to observe good control of the variation in these properties described above in a simple way by varying the composition of the different comonomers used. Finally, with these concepts in mind, biopolymers obtained were tested through assessment of several biological parameters such as cell viability and differentiation with MC3T3-E1 pre-osteoblastic cell line. Results obtained show that these biopolymers maintain good results in terms of cell viability and differentiation, along with tunable properties in terms of hydrophilic character, mechanical properties and enzymatic degradation.

Effect of Gelatin Coating and GO Incorporation on the Properties and Degradability of Electrospun PCL Scaffolds for Bone Tissue Regeneration.

Polymer-based nanocomposites such as polycaprolactone/graphene oxide (PCL/GO) have emerged as alternatives for bone tissue engineering (BTE) applications. The objective of this research was to investigate the impact of a gelatin (Gt) coating on the degradability and different properties of PCL nanofibrous scaffolds fabricated by an electrospinning technique with 1 and 2 wt% GO. Uniform PCL/GO fibers were obtained with a beadless structure and rough surface. PCL/GO scaffolds exhibited an increase in their crystallization temperature (Tc), attributed to GO, which acted as a nucleation agent. Young's modulus increased by 32 and 63% for the incorporation of 1 and 2 wt% GO, respectively, in comparison with neat PCL. A homogeneous Gt coating was further applied to these fibers, with incorporations as high as 24.7 wt%. The introduction of the Gt coating improved the hydrophilicity and degradability of the scaffolds. Bioactivity analysis revealed that the hydroxyapatite crystals were deposited on the Gt-coated scaffolds, which made them different from their uncoated counterparts. Our results showed the synergic effect of Gt and GO in enhancing the multifunctionality of the PCL, in particular the degradability rate, bioactivity, and cell adhesion and proliferation of hGMSC cells, making it an interesting biomaterial for BTE.

New PCL/PEC Blends: In Vitro Cell Response of Preosteoblasts and Human Mesenchymal Stem Cells.

In this study, new blends of PCL/PEC have been prepared in an easy manner by casting with the objective of obtaining new biomaterials to apply to tissue engineering and bone regeneration. The PCL/PEC blends obtained, together with neat polymer blends, were characterized by infrared spectroscopy (FTIR), atomic force microscopy (AFM), scanning electron microscopy (SEM), differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). This full characterization is the key to disentangle the miscibility, which means good compatibility, of the polymer blends used in this work. The addition of increasing amounts of PEC, has shown in the new biomaterials obtained, a remarkable improvement in relation with the mechanical properties (manageable materials) and above all, in terms of an increase in their hydrophilic character with respect to the PCL neat polymer. The improvement of all these properties is reflected in their biological properties. With these thoughts in mind, the blends obtained were tested through the assessment of several biological parameters such as cell viability, proliferation, and differentiation of both the MC3T3-E1 osteoblastic cell line and hMSCs to evaluate their cell response to different polymer membranes aimed at bone tissue regeneration. "In vitro" biocompatibility methods have been chosen rather than in vivo studies due to their lower cost, faster procedure time, and minimum ethical concerns, and because it was the first time that the biological effects of these blends were studied. The results show that the PCL/PEC blends obtained, with tunable properties in terms of hydrophilic character and hydrolytic degradation, may be regarded as good candidates to perform "in vivo" tests and check their real-life applicability for bone regeneration. The polymer acronym (the weight percentage in the sub index) is PCLx/PECy as noted in table one with the summary of compositions.

Chloroaluminate Gel Electrolytes Prepared with Copolymers Based on Imidazolium Ionic Liquids and Deep Eutectic Solvent AlCl3:Urea.

Polymer gel electrolytes (PGEs) have been prepared with copolymers based on imidazolium ionic liquids and the deep eutectic mixture of AlCl3:urea (uralumina) as liquid electrolyte. The copolymers were synthesized by photopolymerization of vinylpirrolidone or methylmethacrylate with imidazolium bis (trifluoromethane sulfonyl) imide (TFSI) ionic liquid monomer and mixed in an increasing range of wt.% with uralumina. The rheology and electrochemical activity of PGEs were highly dependent on the molar ratio of charged groups and copolymer content. Structure of the PGEs was studied by FTIR and Raman spectroscopy and a correlation between interactions polymer/uralumina and changes in speciation of uralumina was established. Despite the low molecular weight of the copolymers, the resulting polymer electrolytes develop elastomeric character associated with the binding ionic species. Although there is room to improve the electrochemical activity, in this study these new gels provide sufficient electroactivity to make them feasible alternatives as electrolytes in secondary aluminum batteries.

JNK Pathway in CNS Pathologies.

The c-Jun N-terminal kinase (JNK) signalling pathway is a conserved response to a wide range of internal and external cellular stress signals. Beside the stress response, the JNK pathway is involved in a series of vital regulatory mechanisms during development and adulthood that are critical to maintain tissue homeostasis. These mechanisms include the regulation of apoptosis, growth, proliferation, differentiation, migration and invasion. The JNK pathway has a diverse functionality and cell-tissue specificity, and has emerged as a key player in regeneration, tumorigenesis and other pathologies. The JNK pathway is highly active in the central nervous system (CNS), and plays a central role when cells need to cope with pathophysiological insults during development and adulthood. Here, we review the implications of the JNK pathway in pathologies of the CNS. More specifically, we discuss some newly identified examples and mechanisms of JNK-driven tumor progression in glioblastoma, regeneration/repair after an injury, neurodegeneration and neuronal cell death. All these new discoveries support the central role of JNK in CNS pathologies and reinforce the idea of JNK as potential target to reduce their detrimental effects.

Preparation and characterization of maleoylagarose/PNIPAAm graft copolymers and formation of polyelectrolyte complexes with chitosan.

A water soluble derivative in 98% yield with 23.1% incorporation of maleoyl groups was obtained by esterification of agarose with maleic anhydride. Graft copolymers were synthesized through vinyl groups of maleoylagarose with N-isopropylacrylamide using ceric ammonium nitrate or ammonium persulfate as initiator, by conventional method or microwave irradiation. High nitrogen content (4.6%) was obtained in the grafting process using ceric ammonium nitrate as initiator without microwave irradiation. Copolymers were characterized by FT-IR and NMR spectroscopies, TGA, DSC and morphological analysis by AFM and SEM microscopy, confirming the grafting of PNIPAAm onto polysaccharide backbone. Hydrogel films were obtained by ionic complexation between opposite charged groups of maleoylagarose-g-poly(N-isopropylacrylamide) and chitosan. The swelling of 1:1w/v maleoylagarose-g-PNIPAAm:chitosan film was higher than 2:1w/v film at 25 and 37°C. 53% release in vitro of diclofenac sodium from 1:1w/v maleoylagarose-g-PNIPAAm:chitosan was obtained at 37°C and pH 6.0 with <0.5 diffusional constant values.

Cervical cancer screening cotesting with cytology and MRNA HPV E6/E7 yields high rates of CIN2+ lesions in young women.

BACKGROUND: European guidelines recommend primary HPV testing for cervical cancer screening. However, the starting age remains to be defined, with an undecided window between 30 and 35 years. This pilot study compares the effectiveness of primary HPV testing to that of cytology for the detection of high-grade (CIN2+) lesions stratified by age. METHODS: Cotesting with LBC cytology and APTIMA® HPV (AHPV) was performed in 5053 women aged 25-65 in an opportunistic screening program in Madrid. AHPV-positive cases were referred to colposcopy and genotyped for HPV16 and 18/45 (AHPV-GT). Results were analyzed stratified in four age groups. RESULTS: 454 cases (9.0%) were AHPV-positive. Women under 35 had a 30.2% CIN2+ rate, compared to 21.9% and 20.4% for women aged 35-44 or 45-54. There was a significant increase (P < .05) in the rate of CIN2+ in AHPV-GT-positive women when compared to that for other HPV types (AHPV-other), being 43.3% versus 15.7%. AHPV-GT-positive women under 35 had significantly higher rates of CIN2+ lesions than any other age-group. The sensitivity of cytology for cervical CIN2+ in APHV-positive women was 60.6%. All 4 carcinomas, including one AHPV-negative endometrial adenocarcinoma, had abnormal cytology. All cervical CIN2+ lesions biopsied were AHPV-positive. CONCLUSIONS: Aptima HPV shows a significantly higher sensitivity for cervical CIN2+ lesions than cytology alone. Unexpectedly, AHPV-positive women under 35 had the highest incidence of CIN2+ lesions, particularly when they are HPV16/18/45-positive. Reconsidering HPV primary screening before the recommended age of 35 is warranted.

Applying the Paris System for Reporting Urine Cytology Increases the Rate of Atypical Urothelial Cells in Benign Cases: A Need for Patient Management Recommendations.

The Paris System (TPS) for reporting urinary cytology attempts to unify the terminology in this field. OBJECTIVES: To analyze the impact of adopting TPS by measuring nomenclature agreement and cytohistological correlation. MATERIALS AND METHODS: Voided urine liquid-based cytology samples corresponding to 149 biopsy-proven cases (76 high-grade carcinomas, 40 low-grade carcinomas, and 33 benign lesions), were reclassified by the same pathologist using TPS. Diagnostic agreement and sensitivity for both nomenclature systems was measured. RESULTS: When using TPS, the rate of atypical samples increased 8 times (from 3 to 24.2%) in benign cases, 10 times (from 2.5 to 25%) in low-grade carcinomas, and 2.4 times (from 6.6 to 15.8%) in high-grade carcinomas. The false-positive rate (abnormal cytology in negative or low-grade carcinoma cases) increased from 11 to 34.2%. Sensitivity was higher (63 vs. 49%) with TPS at the expense of a lower specificity (73 vs. 91%). The agreement between both nomenclatures was moderate for negative and high-grade carcinoma cases (k = 0.42 and 0.56, respectively) and weak for low-grade tumors (k = 0.35). CONCLUSIONS: Adopting TPS for reporting urine cytology results in a considerable increase in atypical diagnoses, improving sensitivity but lowering specificity. Appropriate management recommendations for patients with an atypical cytological diagnosis are required.

Aislamiento y caracterización de células mesenquimales derivadas de tejido adiposo / Isolation and characterization of mesenchymal cells from adipose tissue

Introducción : Existe un creciente interés científico en el potencial terapéutico de las células madre mesenquimales derivadas de tejido adiposo ( ADSCs, en inglés). Estas células son abundantes en el tejido adiposo, son de fácil obtención y con un alto potencial de diferenciación hacia linajes celulares especializados incluyendo adipocitos, osteocitos, condrocitos, miocitos, cardiomiocitos, tenocitos, vasos sanguíneos y neuronas. Este trabajo se desarrolló con el objetivo de implementar en el laboratorio un procedimiento para aislar y cultivar ADSCs, con características que corresponden a las informadas para este linaje celular.Método: los precursores de células adiposas humanas se obtuvieron de tejido subcutáneo abdominal. Las células se separaron enzimáticamente del tejido y se decantaron por centrifugación, luego de cultivadas, se caracterizaron en su capacidad de diferenciación y por su marcadores fenotípicos.Resultados: Las ADSCs aisladas se replicaron en estas condiciones de cultivo y mantuvieron un fenotipo estable durante todo el período de estudio. Se comprobó su potencial adipogénico y osteogénico in vitro, como corresponde a las células madre mesenquimales. El estudio por citometría de flujo mostró que estas células expresan CD73, CD90 y CD105 y son negativas para los marcadores de linaje hematopoyético CD34 y CD45.En los ensayos de inhibición in vitro, las ADSCs demostraron su capacidad para inhibir la proliferación de células T humanas. Conclusiones : La caracterización fenotípica y funcional de las ADSCs obtenidas a partir del tejido adiposo abdominal demuestra que es posible la obtención mediante cultivo in vitro de células mesenquimales humanas sin inducir diferenciación espontánea, manteniendo su integridad funcional y altos niveles de proliferación, lo que sienta las bases para el inicio de ensayos preclínicos y su uso futuro en la terapia celular en nuestro país(AU)

Introduction : There is growing scientific interest in the therapeutic potential of stem cells derived from adipose tissue (ADSCs). These cells are abundant in adipose tissue, are readily available and have a high potential fordifferentiation into specialized cell lineages including adipocytes, osteocytes, chondrocytes, myocytes, cardiomyocytes, tenocyte, endothelial cells and neurons. The aim of this work was to isolate, cultivate andcharacterize ADSCs.Methods : human adipose precursor cells were obtained from abdominal subcutaneous tissue. Cells were enzymatically separated from the tissue and decanted by centrifugation, cultured and finally analyzed.Results : The ADSCs were able to replicate in our culture conditions. The cells maintained their phenotype in different passages throughout the study period, confirming its feasibility for in vitro culture. Also the ADSCs were induced to adipogenic and osteogenic differentiation, verifying its potential as mesenchymal stem cells in vitro. The flow cytometric study showed that these cells expressed CD73, CD90 and CD105 (markers of mesenchymal cells) and they were negative for CD34 and CD45 (hematopoieticcell markers). The ADSCs were able to inhibit in vitro the proliferation of T cells.Conclusions : It is possible to obtain ADSCs by in vitro cultivation without adipogenic induction, maintaining its functional integrity and high proliferation; this cell could be an important tool for the cellular therapy in our country(AU)

Aislamiento y caracterización de células mesenquimales derivadas de tejido adiposo / Isolation and characterization of mesenchymal cells from adipose tissue

Introducción : Existe un creciente interés científico en el potencial terapéutico de las células madre mesenquimales derivadas de tejido adiposo ( ADSCs, en inglés). Estas células son abundantes en el tejido adiposo, son de fácil obtención y con un alto potencial de diferenciación hacia linajes celulares especializados incluyendo adipocitos, osteocitos, condrocitos, miocitos, cardiomiocitos, tenocitos, vasos sanguíneos y neuronas. Este trabajo se desarrolló con el objetivo de implementar en el laboratorio un procedimiento para aislar y cultivar ADSCs, con características que corresponden a las informadas para este linaje celular. Método: los precursores de células adiposas humanas se obtuvieron de tejido subcutáneo abdominal. Las células se separaron enzimáticamente del tejido y se decantaron por centrifugación, luego de cultivadas, se caracterizaron en su capacidad de diferenciación y por su marcadores fenotípicos. Resultados: Las ADSCs aisladas se replicaron en estas condiciones de cultivo y mantuvieron un fenotipo estable durante todo el período de estudio. Se comprobó su potencial adipogénico y osteogénico in vitro, como corresponde a las células madre mesenquimales. El estudio por citometría de flujo mostró que estas células expresan CD73, CD90 y CD105 y son negativas para los marcadores de linaje hematopoyético CD34 y CD45.En los ensayos de inhibición in vitro, las ADSCs demostraron su capacidad para inhibir la proliferación de células T humanas. Conclusiones : La caracterización fenotípica y funcional de las ADSCs obtenidas a partir del tejido adiposo abdominal demuestra que es posible la obtención mediante cultivo in vitro de células mesenquimales humanas sin inducir diferenciación espontánea, manteniendo su integridad funcional y altos niveles de proliferación, lo que sienta las bases para el inicio de ensayos preclínicos y su uso futuro en la terapia celular en nuestro país(AU)

Introduction : There is growing scientific interest in the therapeutic potential of stem cells derived from adipose tissue (ADSCs). These cells are abundant in adipose tissue, are readily available and have a high potential fordifferentiation into specialized cell lineages including adipocytes, osteocytes, chondrocytes, myocytes, cardiomyocytes, tenocyte, endothelial cells and neurons. The aim of this work was to isolate, cultivate andcharacterize ADSCs. Methods : human adipose precursor cells were obtained from abdominal subcutaneous tissue. Cells were enzymatically separated from the tissue and decanted by centrifugation, cultured and finally analyzed. Results : The ADSCs were able to replicate in our culture conditions. The cells maintained their phenotype in different passages throughout the study period, confirming its feasibility for in vitro culture. Also the ADSCs were induced to adipogenic and osteogenic differentiation, verifying its potential as mesenchymal stem cells in vitro. The flow cytometric study showed that these cells expressed CD73, CD90 and CD105 (markers of mesenchymal cells) and they were negative for CD34 and CD45 (hematopoieticcell markers). The ADSCs were able to inhibit in vitro the proliferation of T cells. Conclusions : It is possible to obtain ADSCs by in vitro cultivation without adipogenic induction, maintaining its functional integrity and high proliferation; this cell could be an important tool for the cellular therapy in our country(AU)

Increased risk of malignancy for non-atypical urothelial cell groups compared to negative cytology in voided urine. Morphological changes with LBC.

Liquid-based cytology (LBC) has recently become the preferred method for urine cytology analysis, but differences with conventional cytology (CC) have been observed. The purpose of this study is to analyze these differences and the clinical relevance of non-atypical urothelial cell groups (UCG) in voided urine specimens. Reporting terminology is discussed. Initially, diagnostic categories from 619 LBC and 474 CC samples, reviewed by five different pathologists, were compared (phase 1). Five years after LBC was implemented and applying strict cytologic criteria for UCG diagnosis, 760 samples were analyzed (phase 2) and compared to previous LBC specimens. Diagnostic differences, interobserver variability and clinicopathological correlation with a 6-month follow-up, were analyzed. UCG increased from 6.5% with CC to 20.7% (218%, 3.2 fold, P < 0.0001) with LBC. This difference was not related to interobserver variability. Five years later, the rate of UCG had decreased to 13 2%. While 6% of cases with a negative cytology had urothelial carcinoma (UC) within 6 months of diagnosis, this percentage increased to 15.7% with UCG. The sensitivity of the UCG category for UC was low (30.4%), but the specificity and the negative predictive value (NPV) were high (87.1% and 94%, respectively). LBC increases UCG when compared to CC. This can be corrected with observers experience and using set cytological criteria. Due to its association with carcinoma, the presence of UCG in voided urine should be framed in a diagnostic category other than "negative for malignancy." Diagn. Cytopathol. 2016;44:582-590. © 2016 Wiley Periodicals, Inc.

Hybrid materials: Magnetite-Polyethylenimine-Montmorillonite, as magnetic adsorbents for Cr(VI) water treatment.

Hybrid materials formed by the combination of a sodium rich Montmorillonite (MMT), with magnetite nanoparticles (40 nm, Fe(3)O(4) NPs) coated with Polyethylenimine polymer (PEI 800 g/mol or PEI 25000 g/mol) were prepared. The intercalation of the magnetite nanoparticles coated with PEI among MMT platelets was achieved by cationic exchange. The resulting materials presented a high degree of exfoliation of the MMT sheets and a good dispersion of Fe(3)O(4) NPs on both the surface and among the layers of MMT. The presence of amine groups in the PEI structure not only aids the exfoliation of the MMT layers, but also gives to the hybrid material the necessary functionality to interact with heavy metals. These hybrid materials were used as magnetic sorbent for the removal of hexavalent chromium from water. The effect that pH, Cr(VI) concentration, and adsorbent material composition have on the Cr(VI) removal efficiency was studied. A complete characterization of the materials was performed. The hybrid materials showed a slight dependence of the removal efficiency with the pH in a wide range (1-9). A maximum amount of adsorption capacity of 8.8 mg/g was determined by the Langmuir isotherm. Results show that these hybrid materials can be considered as potential magnetic adsorbent for the Cr(VI) removal from water in a wide range of pH.

Nanocomposite microcontainers.

Versatile all-nanocomposite capped microcontainers are made using layer-by-layer (LBL) assembly. The microcontainers can act as inert packaging with slow/controlled release for virtually any type of encapsulating material based on clay nanocomposites 3D molded by PDMS templates and capped with another LBL film.

Alkaline-treated poly(epsilon-caprolactone) films: degradation in the presence or absence of fibroblasts.

In the first stage, we observed the study of the degradation behavior of alkaline-treated poly(epsilon-caprolactone) (PCL) in two biologically-related media: phosphate buffered saline (PBS) and Dulbecco's modified Eagle's medium (DMEM) for 18 months, finding a much accelerated degradation in the last one. As expected, the degradation in the presence of cells is much pronounced even considering that the study is limited to 6 months. The characterization of the degraded substrates by chemiluminescence (CL) allows to explain the modifications of the substrate and their relations with transitory oxidative stress phenomena described in the fibroblasts seeded onto the PCL membranes.

Fluorescent probes for sensing processes in polymers.

Fluorescence spectroscopy is an important analytical technique that has been widely used in a variety of applications, such as biomedicine, biology, and science of materials, because it presents some properties which makes it unique, that is, extraordinary sensitivity and selectivity, short delay time (<10(-9) s), and it is neither invasive nor destructive, so it can be used for in situ measurements. Generally, intrinsic fluorescence of many materials, like polymers, is unspecific so it is not useful to analyse their properties or to be correlated to changes in their microenvironment. The incorporation of additives with fluorescent groups would be necessary. When the fluorescence emission of these molecules is sensitive to changes of properties, such as polarity, fluidity, order, molecular mobility, pH, or electric potential, they can be used for detecting such changes in their microenvironment, and they are called fluorescent probes. As long as these probes can follow processes of practical interest, they can be employed as sensors, if the information given by the measure of fluorescence adequately reflects the changes in the system. In addition, a sensor must fulfil some other requirements in order to make them of practical use, the most important being that the material support in which the sensor molecule is inserted. This support should permit a rapid detection of the process and should allow easy processing in a variety of forms. Polymers are well-known systems in which estimation of local parameters are possible by means of fluorimetric techniques. It allows the study of dynamic processes of interest, such as polymerization kinetics and mechanisms, thermal transitions, photodegradation, swelling morphology changes, and so forth.