RESUMO
AIM: To assess the diagnostic value of apparent diffusion coefficient (ADC) on 3 T device for the prediction of tumoral response to neoadjuvant chemoradiotherapy (nCRT) and for the response assessment after nCRT in patients with locally advanced rectal cancer (LARC), using pathology as a reference. METHODS: Forty-one patients affected by LARC undergoing 3.0 T MRI before and after nCRT were retrospectively selected. After the conventional acquisition of high resolution T2-weighted sequences, diffusion-weighted MRI (DW-MRI) was performed using a spin-echo echo-planar sequence with multiple b values (150, 500, 1000, 1500 s/mm2 ). Fitted ADC values were calculated for each rectal lesion before and after nCRT by drawing a hand-made region of interest (ROI) around the tumour outline. All patients underwent surgery and pathological staging (classified according to tumour regression grading [TRG] and to tumour and node [TN]) represented the reference standard. Pretreatment ADC value (pre-ADC), ADC value obtained after nCRT (post-ADC) and the difference between post-ADC and pre-ADC (ΔADC) were correlated with both the TRG classes and the TN staging system in each patient. RESULTS: The ADC values obtained in the post nCRT examination and the ΔADC were statistically related both to TRG (p = 0.0004; p = 0.0126, respectively) and TN (p = 0.0484; p = 0.0673, respectively) stages at histopathology. On the contrary, the pre-ADC was not related either to the TRG classes or to the lesion TN staging system (p > 0.05). CONCLUSIONS: 3 T DW-MRI using ADC value can be useful to assess the efficacy of nCRT in LARC; in fact, post-ADC and ΔADC values improve MR capability to evaluate tumour response.
Assuntos
Segunda Neoplasia Primária , Neoplasias Retais , Humanos , Imagem de Difusão por Ressonância Magnética/métodos , Neoplasias Retais/diagnóstico por imagem , Neoplasias Retais/terapia , Neoplasias Retais/patologia , Terapia Neoadjuvante/métodos , Estudos Retrospectivos , Resultado do Tratamento , Imageamento por Ressonância Magnética , QuimiorradioterapiaRESUMO
The 3-D spatial and mechanical features of nano-topography can create alternative environments, which influence cellular response. In this paper, murine fibroblast cells were grown on surfaces characterized by protruding nanotubes. Cells cultured on such nano-structured surface exhibit stronger cellular adhesion compared to control groups, but despite the fact that stronger adhesion is generally believed to promote cell cycle progression, the time cells spend in G1 phase is doubled. This apparent contradiction is solved by confocal microscopy analysis, which shows that the nano-topography inhibits actin stress fiber formation. In turn, this impairs RhoA activation, which is required to suppress the inhibition of cell cycle progression imposed by p21/p27. This finding suggests that the generation of stress fibers, required to impose the homeostatic intracellular tension, rather than cell adhesion/spreading is the limiting factor for cell cycle progression. Indeed, nano-topography could represent a unique tool to inhibit proliferation in adherent well-spread cells.
Assuntos
Adesão Celular , Ciclo Celular , Fibroblastos/fisiologia , Nanoestruturas/química , Animais , Divisão Celular , Células Cultivadas , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Fibroblastos/citologia , Camundongos , Alicerces Teciduais , Proteínas rho de Ligação ao GTP/metabolismo , Proteína rhoA de Ligação ao GTPRESUMO
New inorganic nanostructured matrices for fiber-reinforced composites with enhanced high-temperature stability were developed from alkali aluminosilicate polymers doped with different ultra-high-temperature ceramic (UHTC) particles. The alkali aluminosilicate matrices were synthesized at room temperature with a high SiO2:Al2O3 ratio and then further functionalized by doping with 4-5 wt % of micrometric SiC, ZrB2, ZrC, and HfC powders and finally thermally stabilized as glass-ceramics at 750 °C. The different UHTC-doped matrices were characterized according to their dimensional and microstructural changes after thermal cycling in air flux at 1000 °C. The first results showed that carbide-based UHTC powders improved the thermal stability of the matrices, preventing the excessive swelling of the material and the formation of detrimental voids that might result in the lack of adhesion with reinforcing fibers. Contrarily, the addition of ZrB2 resulted in an excessive matrix swelling at high temperature, thus proving no efficacy compared to the undoped matrix. Impregnation tests carried out on C-fiber fabrics showed good processability, adhesion to the fibers, and fracture pull-out, especially for carbide-based matrices.
RESUMO
The development of advanced materials with biomimetic features in order to elicit desired biological responses and to guarantee tissue biocompatibility is recently gaining attention for tissue engineering applications. Bioceramics, such as hydroxyapatite-based biomaterials are now used in a number of different applications throughout the body, covering all areas of the skeleton, due to their biological and chemical similarity to the inorganic phases of bones. When bioactive sintered hydroxyapatite (HA) is desired, biomolecular modification of these materials is needed. In the present work, we investigated the influence of plasma surface modification coupled to chemical grafting on the cell growth compliance of HA 3D scaffolds.
Assuntos
Divisão Celular , Durapatita/química , Gases em Plasma , Alicerces Teciduais , Materiais Biocompatíveis , Células Cultivadas , Corantes Fluorescentes/química , Humanos , Microscopia Eletrônica de Varredura , Espectrometria de Fluorescência , Propriedades de Superfície , Difração de Raios XRESUMO
In this paper, four near-net shaped foams were produced via direct foaming, starting from a benchmark metakaolin-based geopolymer formulation. Hydrogen peroxide and metallic silicon were used in different amounts as blowing agents to change the porosity from meso- to ultra-macro-porosity. Foams were characterized by bulk densities ranging from 0.34 to 0.66 g cm-3, total porosity from 70% to 84%, accessible porosity from 41% to 52% and specific surface area from 47 to 94 m2 g-1. Gas permeability tests were performed, showing a correlation between the pore features and the processing methods applied. The permeability coefficients k1 (Darcian) and k2 (non-Darcian), calculated applying Forchheimer's equation, were higher by a few orders of magnitude for the foams made using H2O2 than those made with metallic silicon, highlighting the differing flow resistance according to the interconnected porosity. The gas permeability data indicated that the different geopolymer foams, obtained via direct foaming, performed similarly to other porous materials such as granular beds, fibrous filters and gel-cast foams, indicating the possibility of their use in a broad spectrum of applications.
RESUMO
Among the available technologies for ammonium removal from wastewater, ion exchange represents one of the most promising ones in the perspective to recover ammonium and produce a fertilizing product. However, the vast majority of previous studies on ammonium ion exchange did not evaluate the process robustness under real operational conditions nor optimized the desorption step. In this paper, tests of ammonium removal and recovery were conducted on a metakaolin K-based geopolymer, compared with a high-performing Italian natural zeolite in K-form. Real municipal and saline wastewater was treated in a continuous flow pilot plant equipped with a 60-cm adsorption bed (bed volume 203 mL, sorbent mass 145-173 g, empty bed contact time 10 min). Geopolymer granules showed higher performances in terms of selectivity towards ammonium, operating capacity (8.5 mgN g-1 dry adsorbent at an inlet concentration of 40 mgN L-1), bed volumes of wastewater treated at the selected breakpoint (149). Geopolymer resulted to be a cost-effective adsorbent for wastewater treatment capable to adsorb cations by ion exchange, allowing a fractionated desorption procedure that led to recover ammonium in a solution composed mainly by NH4NO3 (37%wt) and KNO3 (56%wt), potentially usable as fertilizer. The geopolymer robustness was assessed after repeated adsorption/regeneration cycles showing that the geopolymer mechanical and morphological properties did not deteriorate. The results make the tested geopolymer a very promising material for the optimization and scale-up of the ammonium recovery process in a circular economy perspective.
Assuntos
Compostos de Amônio , Zeolitas , Águas Residuárias , Troca Iônica , Fertilizantes , AdsorçãoRESUMO
Neuronal CIPP (channel-interacting PDZ protein) is a multivalent PDZ protein that interacts with specific channels and receptors highly expressed in the brain. It is composed of four PDZ domains that behave as a scaffold to clusterize functionally connected proteins. In the present study, we selected a set of potential CIPP interactors that are involved directly or indirectly in mechanisms of cytoskeletal remodelling and membrane protrusion formation. For some of these, we first proved the direct binding to specific CIPP PDZ domains considered as autonomous elements, and then confirmed the interaction with the whole protein. In particular, the small G-protein effector IRSp53 (insulin receptor tyrosine kinase substrate protein p53) specifically interacts with the second PDZ domain of CIPP and, when co-transfected in cultured mammalian cells with a tagged full-length CIPP, it induces a marked reorganization of CIPP cytoplasmic localization. Large punctate structures are generated as a consequence of CIPP binding to the IRSp53 C-terminus. Analysis of the puncta nature, using various endocytic markers, revealed that they are not related to cytoplasmic vesicles, but rather represent multi-protein assemblies, where CIPP can tether other potential interactors.
Assuntos
Proteínas de Transporte/metabolismo , Citoesqueleto/metabolismo , Animais , Western Blotting , Encéfalo/metabolismo , Proteínas de Transporte/genética , Linhagem Celular , Cromatografia de Afinidade , Citosol/metabolismo , Humanos , Imunoprecipitação , Camundongos , Microscopia de Fluorescência , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/fisiologia , Ligação Proteica , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Geopolymer beads, conceived as alternative low cost adsorbents for wastewater treatment, were shaped by a dripping technique in liquid nitrogen, through an ice-templating process. PEG600 was added as a binder to ease the process, standardizing the beads dimension. The beads were investigated in terms of morphology, microstructure and mechanical strength, following compressive tests by ISO 18591. Functional tests, to verify the adsorption capacity, were conducted using methylene blue (MB) with different concentrations and for different contact time. The removal efficiency was mainly related to the morphology and porosity of the beads, which in turn was directly related to the water content added to the geopolymer slurry. In general, all beads reached an average removal efficiency of 98% after 24 h. However, the best performing beads were able to uptake MB very quickly, attaining a removal efficiency of 76% after only 30 min.
RESUMO
With the ultimate goal to generate suitable materials for the repair of osteochondral defects, in this work we aimed at developing composite osteochondral scaffolds organized in different integrated layers, with features which are biomimetic for articular cartilage and subchondral bone and can differentially support formation of such tissues. A biologically inspired mineralization process was first developed to nucleate Mg-doped hydroxyapatite crystals on type I collagen fibers during their self-assembling. The resulting mineral phase was non-stoichiometric and amorphous, resembling chemico-physical features of newly deposited, natural bone matrix. A graded material was then generated, consisting of (i) a lower layer of the developed biomineralized collagen, corresponding to the subchondral bone, (ii) an upper layer of hyaluronic acid-charged collagen, mimicking the cartilaginous region, and (iii) an intermediate layer of the same nature as the biomineralized collagen, but with a lower extent of mineral, resembling the tidemark. The layers were stacked and freeze-dried to obtain an integrated monolithic composite. Culture of the material for 2 weeks after loading with articular chondrocytes yielded cartilaginous tissue formation selectively in the upper layer. Conversely, ectopic implantation in nude mice of the material after loading with bone marrow stromal cells resulted in bone formation which remained confined within the lower layer. In conclusion, we developed a composite material with cues which are biomimetic of an osteochondral tissue and with the capacity to differentially support cartilage and bone tissue generation. The results warrant testing of the material as a substitute for the repair of osteochondral lesions in orthotopic animal models.
Assuntos
Materiais Biomiméticos , Cartilagem Articular/metabolismo , Condrócitos/fisiologia , Engenharia Tecidual/métodos , Alicerces Teciduais , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/metabolismo , Cartilagem Articular/química , Cartilagem Articular/citologia , Condrócitos/citologia , Colágeno/química , Colágeno/metabolismo , Humanos , Ácido Hialurônico/química , Ácido Hialurônico/metabolismo , Teste de Materiais , CamundongosRESUMO
Sr and CO3 co-substituted hydroxyapatite (SrCHA) nanopowder was synthesized by neutralization. The powder was characterized. The improved solubility in Hanks' balanced solution of SrCHA granules (400-600 microm of dimensional range), potentially usable as bone filler, was assessed and compared with that of an analogous carbonate free granulate. SrCHA porous bodies with interconnected micro- and macro-porosity, which mimic the morphology of spongy bone, were prepared by the impregnation of cellulose sponges with suspensions of the SrCHA powder and controlled sintering. SrCHA porous scaffolds sintered at 850 degrees C, in flowing CO2 atmosphere, showed satisfying compressive strength (4.58+/-0.75 MPa) for a porosity value of 45 vol.% and retained the desired ionic substitutions (Sr/Ca=0.11 and CO3=6.8 wt.%). The possibility of widely modulating, by acting on the chemical-physical-geometrical features of the material, the prolonged in situ release of therapeutic Sr, together with the fundamental (Ca, PO4) and main substituting (CO3) ions that constitute the bone mineral phase, makes the use of SrCHA as resorbable bone filler or bone substitute scaffolds promising, especially when pathologies related with Sr deficiency are present. In vitro and in vivo tests are in progress.
Assuntos
Carbonatos/química , Durapatita/síntese química , Teste de Materiais , Estrôncio/química , Força Compressiva , Análise Diferencial Térmica , Concentração de Íons de Hidrogênio , Microscopia Eletrônica de Varredura , Porosidade , Pós/síntese química , Solubilidade , Espectroscopia de Infravermelho com Transformada de Fourier , Condutividade Térmica , Termogravimetria , Difração de Raios XRESUMO
Porous apatites, which during resorption can release in situ Sr ions, were prepared to associate an anti-osteoporotic action with the peculiar features of the inorganic phase constituting the bone. Sr-substituted hydroxyapatite (SrHA) powder was directly synthesized using the classical neutralization route, but including Sr ions, and characterized. The higher solubility of SrHA granules of 400-600 microm size, potentially usable as a bone filler, was assessed compared with that of analogous stoichiometric HA granules. The Sr released in synthetic body fluid became constant after 1 week. The Ca release is improved for SrHA compared with stoichiometric HA, due to the higher solubility of the first material. Porous scaffolds with micro-macro interconnected porosity, which mimic the morphology of the spongy bone, were prepared by the impregnation of cellulose sponges with suspensions of the powder and a specific sintering process. A compressive strength of 4.52+/-1.40 MPa was obtained for SrHA scaffolds characterized with 45 vol.% of porosity. Promising biomedical applications, such as resorbable bone filler or bone substitute releasing in situ Sr ions for a prolonged time, can be hypothesized for the SrHA materials when pathologies related with Sr deficiency are present.
Assuntos
Substitutos Ósseos/química , Hidroxiapatitas/química , Osteoporose , Estrôncio/química , Cálcio/química , Simulação por Computador , Concentração de Íons de Hidrogênio , Teste de Materiais , Microscopia Eletrônica de Varredura , Osteoporose/cirurgia , Porosidade , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios XRESUMO
PDZ domains are protein-protein interaction modules that are crucial for the assembly of structural and signalling complexes. They specifically bind to short C-terminal peptides and occasionally to internal sequences that structurally resemble such peptide termini. The binding of PDZ domains is dominated by the residues at the P(0) and P(-2) position within these C-terminal targets, but other residues are also important in determining specificity. In this study, we analysed the binding specificity of the third PDZ domain of protein tyrosine phosphatase BAS-like (PTP-BL) using a C-terminal combinatorial peptide phage library. Binding of PDZ3 to C-termini is preferentially governed by two cysteine residues at the P(-1) and P(-4) position and a valine residue at the P(0) position. Interestingly, we found that this binding is lost upon addition of the reducing agent dithiothrietol, indicating that the interaction is disulfide-bridge-dependent. Site-directed mutagenesis of the single cysteine residue in PDZ3 revealed that this bridge formation does not occur intermolecularly, between peptide and PDZ3 domain, but rather is intramolecular. These data point to a preference of PTP-BL PDZ3 for cyclic C-terminal targets, which may suggest a redox state-sensing role at the cell cortex.
Assuntos
Cisteína/metabolismo , Fragmentos de Peptídeos/metabolismo , Proteínas Tirosina Fosfatases/metabolismo , Motivos de Aminoácidos , Sequência de Aminoácidos , Animais , Sítios de Ligação , Células COS , Chlorocebus aethiops , Técnicas de Química Combinatória , Cisteína/química , Cisteína/genética , Glutationa Transferase/metabolismo , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Oxirredução , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/genética , Biblioteca de Peptídeos , Plasmídeos , Ligação Proteica , Conformação Proteica , Estrutura Terciária de Proteína , Proteína Tirosina Fosfatase não Receptora Tipo 13 , Proteínas Tirosina Fosfatases/química , Proteínas Tirosina Fosfatases/genética , Proteínas Recombinantes de Fusão/metabolismo , Homologia de Sequência de AminoácidosRESUMO
The effectiveness of synthetic body fluids (SBF) as biomimetic sources to synthesize carbonated hydroxyapatite (CHA) powder similar to the biological inorganic phase, in terms of composition and microstructure, was investigated. CHA apatite powders were prepared following two widely experimented routes: (1) calcium nitrate tetrahydrate and diammonium hydrogen phosphate and (2) calcium hydroxide and ortophosphoric acid, but using SBF as synthesis medium instead of pure water. The characteristics of the as-prepared powders were compared, also with the features of apatite powders synthesized via pure water-based classical methods. The powder thermal resistance and behaviour during densification were studied together with the mechanical properties of the dense samples. The sponge impregnation process was used to prepare porous samples having morphological and mechanical characteristics suitable for bone substitution. Using this novel synthesis was it possible to prepare nanosized (approximately equal to 20 nm), pure, carbonate apatite powder containing Mg, Na, K ions, with morphological and compositional features mimicking natural apatite and with improved thermal properties. After sintering at 1250 degrees C the carbonate-free apatite porous samples showed a surprising, high compressive strength together with a biomimetic morphology.
Assuntos
Apatitas/química , Materiais Biocompatíveis/química , Biomimética , Líquidos Corporais/química , Substitutos Ósseos , Carbono/química , Durapatita/química , Compostos de Cálcio/química , Hidróxido de Cálcio/química , Carbonatos/química , Concentração de Íons de Hidrogênio , Íons , Magnésio/química , Teste de Materiais , Microscopia Eletrônica de Varredura , Modelos Químicos , Nitratos/química , Fosfatos/química , Ácidos Fosfóricos/química , Potássio/química , Pós , Sódio/química , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Temperatura , Água/química , Difração de Raios XRESUMO
A new method to synthesise carbonate-substituted hydroxyapatite (CHA) powder has been set up introducing a CO(2) flux, as a source of carbonate, in the HA synthesis process based on the neutralisation reaction. The reactants are abundant and inexpensive. The yield is good compared to other CHA powder synthesis. The reaction may be performed at low temperature and without pH control and does not produce any by-products. The influence of the synthesis parameters (temperature, H(3)PO(4) solution dropping rate, i.e. reaction time, CO(2) flux, ageing time) has been tested to optimise the process conditions in order to obtain the highest carbonation degree and favour the B-type CHA precipitation with respect to A-type one. The prepared powder (5.8wt% of total carbonate with an A/B ratio of 0.78) was thermally treated at various temperatures in the range 500-1400 degrees C in different atmospheres (air, wet and dry carbon dioxide). The thermal treatments were performed with a double aim, to eliminate selectively the carbonate groups in A-position maintaining the B-type substitution, and to evaluate the thermal stability of the CHA and the total loss of carbonate as a function of temperature. The thermal treatment at 900 degrees C in wet CO(2) gave the best result in terms of a high carbonate residue and a low A/B ratio. We also investigate the use of different techniques (inductively coupled plasma, TGA, Fourier transformed infrared spectroscopy, X-ray diffraction) for characterising CHA and calculating sensitivity and accuracy in the quantification of carbonate ions for each molecular site.
Assuntos
Apatitas/química , Materiais Biocompatíveis/química , Temperatura Alta , Teste de Materiais/métodos , Apatitas/isolamento & purificação , Apatitas/efeitos da radiação , Materiais Biocompatíveis/síntese química , Dióxido de Carbono/química , Conformação Molecular , Tamanho da Partícula , Ácidos Fosfóricos/química , Pós , Propriedades de SuperfícieRESUMO
Replacement of bone tissue by graft materials and products of tissue engineering having composition, structure, and biological features that mimic natural tissue is a goal to be pursued. A biomimetic synthesis was performed to prepare new bone-like composites constituted of hydroxyapatite nanocrystals and self-assembled type I collagen fibers. We used a biological inspired approach that proved that the biological systems stored and processed information at the molecular level. Two different methodologies were used: dispersion of synthetic hydroxyapatite in telopeptides free collagen molecules solution and direct nucleation of hydroxyapatite into reconstituted collagen fibers during their assembling. The different preparation techniques were experimented then the composites thoroughly characterized and compared. Composite obtained by direct nucleation showed an intimated interaction of the inorganic and proteic components, which modified the apatitic phase and made its composition, morphology and structure similar to the mineral component of natural bone.
Assuntos
Substitutos Ósseos/síntese química , Colágeno/química , Durapatita/química , Substitutos Ósseos/química , Colágeno/ultraestrutura , Temperatura Alta , Microscopia Eletrônica de Varredura , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios XRESUMO
Porous macro-granules of nanostructured apatite with Ca ions partially cosubstituted with Mg and Sr ions in different ratios (SrMgHAs), were synthesized at 37°C and compared with Mg and/or Sr free apatites (MgHAs and HA). Strontium improved the Mg substitution extent in the apatite and the chemical-physical and thermal stability of the resulting cosubstituted apatite. Porous macro-granules of 400-600 micron with selected composition were tested for the ionic release in synthetic body fluid and the data were related with the results of preliminary cell investigation in vitro. As compared to the corresponding Sr-free granulate, the SrMgHA could be exploited to prolong the beneficial Mg release during the bone regeneration process. In addition the contemporary in situ supply of Sr, an antiosteoporotic and anticarie ion, could influence the quality of new hard tissues. The ionic multirelease created a more favorable environment for human osteoblasts, demonstrated by a proliferative effect for each dose tested in the range 0.1-10 mg/mL.
Assuntos
Substitutos Ósseos/química , Durapatita/química , Regeneração Óssea/efeitos dos fármacos , Substitutos Ósseos/administração & dosagem , Substitutos Ósseos/farmacologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Durapatita/administração & dosagem , Durapatita/farmacologia , Humanos , Magnésio/química , Teste de Materiais , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoporose/terapia , PorosidadeRESUMO
In this study, rapid CAD-CAM prototyping of pure hydroxyapatite to replace temporomandibular joint condyles was tested in sheep. Three adult animals were implanted with CAD-CAM-designed porous hydroxyapatite scaffolds as condyle substitutes. The desired scaffold shape was achieved by subtractive automated milling machining (block reduction). Custom-made surgical guides were created by direct metal laser sintering and were used to export the virtual planning of the bone cut lines into the surgical environment. Using the same technique, fixation plates were created and applied to the scaffold pre-operatively to firmly secure the condyles to the bone and to assure primary stability of the hydroxyapatite scaffolds during masticatory function. Four months post-surgery, the sheep were sacrificed. The hydroxyapatite scaffolds were explanted, and histological specimens were prepared. Different histological tissues penetrating the scaffold macropores, the sequence of bone remodeling, new apposition of bone and/or cartilage as a consequence of the different functional anatomic role, and osseointegration at the interface between the scaffold and bone were documented. This animal model was found to be appropriate for testing CAD-CAM customization and the biomechanical properties of porous, pure hydroxyapatite scaffolds used as joint prostheses.
Assuntos
Substitutos Ósseos/uso terapêutico , Durapatita/uso terapêutico , Côndilo Mandibular/cirurgia , Ovinos/cirurgia , Alicerces Teciduais/química , Animais , Substitutos Ósseos/química , Desenho Assistido por Computador , Durapatita/química , Côndilo Mandibular/patologia , Porosidade , Procedimentos de Cirurgia Plástica/métodosRESUMO
We report the identification of a novel partner of Kidins220/ARMS (Kinase D-interacting substrate of 220 kDa/Ankyrin Repeat-rich Membrane Spanning) an adaptor of neurotrophin receptors playing crucial roles during neurogenesis. Screening a phage display library of brain cDNA products we found that D. rerio Pdzrn3, a protein containing RING-finger and PDZ-domains, interacts with Kidins220/ARMS through its first PDZ-domain. Both zebrafish proteins share high homology with the corresponding mammalian proteins and both genes are developmentally expressed in neural districts where early neurogenesis occurs. The interaction was also confirmed by biochemical assays and by co-localization at the tips of growing neurites of PC12 cells induced with nerve growth factor.
Assuntos
Proteínas de Transporte/química , Proteínas de Transporte/metabolismo , Proteínas de Membrana/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Sequência de Aminoácidos , Animais , Biologia Computacional , Células HEK293 , Humanos , Proteínas de Membrana/química , Camundongos , Dados de Sequência Molecular , Proteínas do Tecido Nervoso/química , Células PC12 , Ligação Proteica , Estrutura Terciária de Proteína , Ratos , Ubiquitina-Proteína Ligases , Proteínas de Peixe-Zebra/química , Proteínas de Peixe-Zebra/metabolismoRESUMO
The use of magnetic activation has been proposed to answer the growing need for assisted bone and vascular remodeling during template/scaffold regeneration. With this in mind, a synthesis procedure was developed to prepare bioactive (Fe2+/Fe3+)-doped hydroxyapatite (Fe-HA), endowed with superparamagnetic-like properties. This new class of magnetic hydroxyapatites can be potentially employed to develop new magnetic ceramic scaffolds with enhanced regenerative properties for bone surgery; in addition, magnetic Fe-HA can find application in anticancer therapies, to replace the widely used magnetic iron oxide nanoparticles, whose long-term cytotoxicity was recently found to reach harmful levels. An extensive physicochemical, microstructural and magnetic characterization was performed on the obtained Fe-HA powders, and demonstrated that the simultaneous addition of Fe2+ and Fe3+ ions during apatite nucleation under controlled synthesis conditions induces intrinsic magnetization in the final product, minimizing the formation of magnetite as secondary phase. This result potentially opens new perspectives for biodevices aimed at bone regeneration and for anti-cancer therapies based on hyperthermia.
Assuntos
Materiais Biocompatíveis/química , Durapatita/química , Temperatura Alta , Ferro/química , Magnetismo , Animais , Materiais Biocompatíveis/farmacologia , Adesão Celular/efeitos dos fármacos , Durapatita/farmacologia , Ferro/farmacologia , Campos Magnéticos , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Oxirredução/efeitos dos fármacos , Pós , Coelhos , Espectroscopia por Absorção de Raios X , Difração de Raios XRESUMO
An efficient method for the direct and covalent decoration of granules of nanostructured apatite with a sample monosaccharide is presented; the hydroxyapatite material was directly functionalised with a short azido-containing spacer arm, to which α-propargyl glucopyranoside has been chemoselectively ligated by Huisgen-type cycloaddition. The 'glycosylated' hydroxypatite was characterised by its ability to interact with glucose recognising lectins.