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Porous structures are essential for some collagen-based biomaterials and can be regulated by crosslinkers. Herein, dialdehyde carboxymethyl cellulose (DCMC) crosslinkers with similar size but different aldehyde group contents were prepared through periodate oxidation of sodium carboxymethyl cellulose with varying degrees of substitution (DS). They can penetrate into the hierarchy of fibril and form inter-molecular and intra-fibril cross-linking within the collagen matrix due to their nanoscale sizes and reactive aldehyde groups. The collagen matrices possessed higher porosity, significantly greater proportion of large pores (Φ > 10 µm), and shorter D-periodicity after cross-linking, showing greater potential for biomedical applications. In addition, the crosslinked collagen matrices showed satisfactory biocompatibility and biodegradation. The decreased DS of carboxymethyl cellulose, which led to the increased aldehyde content of corresponding DCMC, brought about an enhanced cross-linking degree, porosity, and proportion of large pores of the crosslinked collagen matrix. DCMC dosage of 6% was sufficient for cross-linking and pore formation. Excess DCMC would physically deposit in the matrix and decrease the porosity instead. Therefore, the desired pore properties of the collagen matrix could be obtained by regulating the structure of DCMC and thereby achieving the required functions of the biomaterial.
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Carboximetilcelulosa de Sodio , Colágeno , Aldehídos , Materiales Biocompatibles/química , Carboximetilcelulosa de Sodio/química , Colágeno/química , Reactivos de Enlaces Cruzados/química , PorosidadRESUMEN
Unraveling the interaction mechanisms of type I collagen with various inorganic nanoparticles is of pivotal importance to construct collagen-based bionanocomposites with hierarchical structures for biomedical, pharmaceutical, and other industrial applications. In this study, synthetic two-dimensional Laponite nanoplatelets (LAP NPs) are surface-functionalized with tetrakis(hydroxymethyl) phosphonium sulfate (THPS) for reinforcing their incorporation with type I collagen matrix by focusing on the influences of the interactions on the hierarchical structures of the collagen. Our results indicate that the LAP NPs can be successfully surface-functionalized with THPS via covalent bonds between the amine-functionalized NPs and the hydroxymethyl groups of THPS. Moreover, the resulting NPs can be well dispersed into the collagen matrix and evenly bound onto the collagen fiber strands and between the collagen fibrils, preserving the native D-periodic banding patterns of the collagen fibrils. The formation of covalent and hydrogen bonds between the collagen and the functionalized NPs can stabilize the intrinsic triple-helical conformation of the collagen, conferring the resulting collagen-based nanocomposites with improved thermal stability and enhanced mechanical properties. We anticipate that a fundamental understanding of the interactions between the collagen and functionalized inorganic nanoparticles would contribute to the design, fabrication, and further application of hierarchical collagen-based bionanocomposites with multifunctions.
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Colágeno Tipo I , Nanocompuestos , Arcilla , SilicatosRESUMEN
In this study, we report the dependence of the nanoparticle dispersion on the zero-conversion initiator efficiency in the nanocomposites formed by poly(N-vinyl carbazole) (PNVK) and acrylic acid-modified iron oxide (AA-Fe3O4) nanoparticles via free radical solution polymerization of the precursor solution, that is, a thorough mixture of 28.5 wt% AA-Fe3O4 nanoparticles and the N-vinyl carbazole (NVK) monomer with the solvent dimethylformamide and azobisisobutyronitrile as an initiator. Here three different types of the dispersion state of AA-Fe3O4 nanoparticles in the PNVK matrix have been distinguished by a combined approach of transmission electron microscopy and small-angle X-ray scattering coupled with real-space models of the nanoparticle assemblies. When the polymerization proceeded with a higher zero-conversion initiator efficiency (f°) by pre-polymerization at 115 °C, the generation of a large amount of free radicals could efficiently induce the dominant surface-initiated polymerization of the NVK monomer with the vinyl groups of tethered acrylic acids; in this case, the constitution of "shorter multiple grafted PNVK chains" threaded AA-Fe3O4 nanoparticles to form particle branches and the branches were joined together from branching points along each branch, thereby forming the network structure. However, once the polymerization was conducted at a lower f° by pre-polymerization at 75 °C, a significant reduction in the generation of free radicals likely greatly reduced the efficiency in the occurrence of surface-initiated polymerization at particle surfaces; nevertheless, the self-polymerization of the NVK monomer could still take place to induce a local demixing between the polymerizing longer PNVK chains and AA-Fe3O4 nanoparticles via the attractive depletion mechanism, thus locally leading to the formation of small aggregates. While if the f° was controlled to be intermediate by polymerization at 100 °C, an optimal balance between the rates of the surface-initiated polymerization and the self-polymerization induced a collective construction built from the network and aggregate structures, exhibiting the structural characteristics of large aggregates. Furthermore, the magnetic coercivity of PNVK/AA-Fe3O4 nanocomposites was found to depend on the dispersion state of the AA-Fe3O4 nanoparticles, presenting a tendency towards enhanced coercivity as the dispersion state changed from large aggregates to small aggregates to network structure.
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Electrostatic compaction of double stranded DNA induced by a positively charged poly(amidoamine) (PAMAM) dendrimer of generation four (G4) was found to produce two unique types of DNA mesophases, in which the DNA bent into superhelices packed in a tetragonal or hexagonal lattice. The structure formed at a lower dendrimer charge density was three-dimensionally (3D) ordered, as characterized by the P41212 space group with a 41 screw axis in a tetragonal arrangement, showing that the weakly bent DNA superhelices with a pitch length of ca. 5.0 nm possessed both identical handedness and phase conservation. The 3D ordered structure transformed into a 2D mesophase at a higher dendrimer charge density, wherein the strongly bent superhelices with a pitch length of ca. 4.0 nm organized in a hexagonal lattice without lateral coherence of helical trajectory. The counterion valency of the protonic acid that is used to charge the dendrimer was found to influence the phase diagram. Under a given dendrimer charge density, the complex with a multivalent acid-protonated dendrimer tended to form structures with less curved DNA, attesting that the driving force of charge matching was reduced by increasing the counterion valency of the dendrimer.
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Dendrímeros , Cristales Líquidos , ADN , ADN Superhelicoidal , Electricidad EstáticaRESUMEN
Collagen is an important biomacromolecule, making up the majority of the extracellular matrix in animal tissues. Naturally occurring crosslinks in collagen stabilize its intermolecular structure in vivo, whereas chemical treatments for introducing synthetic crosslinks are often carried out ex vivo to improve the physical properties or heat stability of the collagen fibres for applications in biomaterials or leather production. Effective protection of intrinsic natural crosslinks as well as allowing them to contribute to collagen stability together with synthetic crosslinks can reduce the need for chemical treatments. However, the contribution of these natural crosslinks to the heat stability of collagen fibres, especially in the presence of synthetic crosslinks, is as yet unknown. Using synchrotron small-angle X-ray scattering, the in situ role of natural and synthetic crosslinks on the stabilization of the intermolecular structure of collagen in skins was studied. The results showed that, although natural crosslinks affected the denaturation temperature of collagen, they were largely weakened when crosslinked using chromium sulfate. The development of synergistic crosslinking chemistries could help retain the intrinsic chemical and physical properties of collagen-based biological materials.
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Colágeno/efectos de la radiación , Sincrotrones , Animales , Bovinos , Compuestos de Cromo/química , Reactivos de Enlaces Cruzados/química , Estructura Molecular , Dispersión del Ángulo Pequeño , Sulfatos/química , Rayos XRESUMEN
We present a mechanism for the selectivity of covalent/electrostatic binding of the Cr(III) ion to collagen, mediated by the kosmotropicity of the anions. Although a change in the long-range ordered structure of collagen is observed after covalent binding (Cr(III)-OOC) in the presence of SO4 2- at pH 4.5, the νsym (COO- ) band remains intense, suggesting a relatively lower propensity for the Cr(III) to bind covalently instead of electrostatically through Cr(H2 O)6 3+ . Replacing SO4 2- with Cl- reduces the kosmotropic effect which further favors the electrostatic binding of Cr(III) to collagen. Our findings allow a greater understanding of mechanism-specific metal binding in the collagen molecule. We also report for the first time, surface-enhanced Raman spectroscopy to analyze binding mechanisms in collagen, suggesting a novel way to study chemical modifications in collagen-based biomaterials.
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Compuestos de Cromo/química , Colágeno/química , Animales , Aniones/química , Fenómenos Biofísicos , Bovinos , Colágeno/metabolismo , Concentración de Iones de Hidrógeno , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Espectrometría Raman/métodosRESUMEN
The conformation of polyelectrolytes in the solution state has long been of interest in polymer science. Herein we utilize all atom molecular dynamics simulations (MD) and small-angle x-ray scattering experiments (SAXS) to elucidate the molecular structure of the model polyelectrolyte homogalacturonan. Several degrees of polymerization were studied and in addition partial methylesterification of the otherwise charge-carrying carboxyl groups was used in order to generate samples with varying intra-chain charge distributions. It is shown that at length scales above around 1nm the conformation of isolated chains has surprisingly little dependence on the charge distribution or the concentration of attendant monovalent salts, reflective of the intrinsic stiffness of the saccharide rings and the dynamical constraints of the glycosidic linkage. Indeed the conformation of isolated chains over all accessible length scales is well described by the atomic coordinates available from fibre diffraction studies. Furthermore, in more concentrated systems it is shown that, after careful analysis of the SAXS data, the form of the inter-particle effects heralded by the emergence of a so-called polyelectrolyte peak, can be extracted, and that this phenomena can be reproduced by multiple chain MD simulations.
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Polysaccharide gels assembled from the anionic biopolymers pectin and carrageenan have been studied using transmission electron microscopy (TEM). Gels were formed in several different ways: for pectin, hydrogen bonding was used to form junction zones between strands, whereas for carrageenan systems, several different ion types were used to form ionotropic networks. Using this approach, several distinct network architectures were realized. In addition to preparing gelled samples for electron microscopy, a set of samples was taken without performing the additional treatment necessitated by the TEM measurements, and these were studied directly by small-angle X-ray scattering (SAXS). Taking careful consideration of the relative merits of different image sizes and available processing techniques, the real-space images acquired by TEM were used via radial integration of the Fourier transform to produce simulated scattering patterns. These intensity-versus-wavevector plots were compared with the results of SAXS experiments carried out on the unadulterated gels using synchrotron radiation. Although information regarding chain thicknesses and flexibilities was found to be modified by labeling and changes in the dielectric constant and mechanical properties of the surroundings in the TEM, the studies carried out here show that careful protocols can produce data sets where information acquired above â¼20 nm is broadly consistent with that obtained by SAXS studies carried out on unadulterated samples. The fact that at larger length scale the structure of these water-rich networks seems largely preserved in the TEM samples suggests that three-dimensional (3D) TEM tomography experiments carried out with careful sample preparation will be valuable tools for measuring network architecture and connectivity; information that is lost in SAXS owing to the intrinsic averaging nature of the technique.
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Microscopía Electrónica de Transmisión , Polisacáridos/química , Polisacáridos/ultraestructura , Difracción de Rayos XRESUMEN
Self-assembled hydrogen-bonded networks of the polysaccharide pectin, a mechanically functional component of plant cell walls, have been of recent interest as biomimetic exemplars of physical gels, and the microrheological and strain-stiffening behaviors have been previously investigated. Despite this detailed rheological characterization of preformed gels, little is known about the fundamental arrangement of the polymers into cross-linking junction zones, the size of these bonded regions, and the resultant network architecture in these hydrogen-bonded materials, especially in contrast to the plethora of such information available for their well-known calcium-assembled counterparts. In this work, in concert with pertinent rheological measurements, an in-depth structural study of the hydrogen-bond-mediated gelation of pectins is provided. Gels were realized by using glucona-delta-lactone to decrease the pH of solutions of pectic polymers that had a (blockwise) low degree of methylesterification. Small-angle X-ray scattering and transmission electron microscopy were utilized to access structural information on length scales on the order of nanometers to hundreds of nanometers, while complementary mechanical properties were measured predominantly using small amplitude oscillatory shear rheology.
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Concentración de Iones de HidrógenoRESUMEN
Frustrated, out-of-equilibrium materials have been of considerable interest for some time and continue to be some of the least understood materials. Recent measurements have shown that many gelled biopolymer materials display slow dynamics on timescales greater than one second, that are not accessible with typical methods, and are characteristic of glassy trapped systems. In this study we have controlled the fine structure of the anionic polysaccharide pectin in order to construct a series of ionotropic gels having differing binding energies between the constituent chains, in an attempt to further understand the slow dynamical processes occurring. Using multi-speckle light scattering techniques it is shown that the slow dynamics observed in these gelled systems are stress-driven. As the binding lengths, and thus the binding energies, of the junction zones between the polymer chains in these networks increase the long-time dynamics initially slow, as might be expected, until a critical level of internal stress is reached upon which the dynamics increase significantly, with gentle creaking punctuated by localised stress-relieving quakes.
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The aberrant fibrillization of huntingtin exon 1 (Httex1) characterized by an expanded polyglutamine (polyQ) tract is a defining feature of Huntington's disease, a neurodegenerative disorder. Recent investigations underscore the involvement of a small EDRK-rich factor 1a (SERF1a) in promoting Httex1 fibrillization through interactions with its N terminus. By establishing an integrated approach with size-exclusion-column-based small- and wide-angle X-ray scattering (SEC-SWAXS), NMR, and molecular simulations using Rosetta, the analysis here reveals a tight binding of two NT17 fragments of Httex1 (comprising the initial 17 amino acids at the N terminus) to the N-terminal region of SERF1a. In contrast, examination of the complex structure of SERF1a with a coiled NT17-polyQ peptide (33 amino acids in total) indicates sparse contacts of the NT17 and polyQ segments with the N-terminal side of SERF1a. Furthermore, the integrated SEC-SWAXS and molecular-simulation analysis suggests that the coiled NT17 segment can transform into a helical conformation when associated with a polyQ segment exhibiting high helical content. Intriguingly, NT17-polyQ peptides with enhanced secondary structures display diminished interactions with SERF1a. This insight into the conformation-dependent binding of NT17 provides clues to a catalytic association mechanism underlying SERF1a's facilitation of Httext1 fibrillization.
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Recent molecular dynamics simulations, verified experimentally by solution-state x-ray scattering experiments, have found that κ-carrageenan chains contain helical secondary structure, akin to that found in the solid-state, even in aqueous solution. Furthermore, upon the addition of ions to single chains the simulations found no evidence that any conformational transitions take place. These findings challenge the long-held assumption that the so-called disorder-to-order transition in carrageenan systems involves a uni-molecular 'coil-to-helix transition'. Herein, the results of further molecular dynamics simulations undertaken using pairs of κ-carrageenan chains in 0.1 M NaI solutions are reported, and are validated experimentally using state-of-the-art solution-state WAXS experiments. From initially separated chains double-helices are shown to form, leading the authors to propose 'two single helices-to-stabilized double-helix' as a description of the molecular events taking place during the disorder-to-order transition.
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Simulación de Dinámica Molecular , Carragenina/química , Rayos X , Conformación MolecularRESUMEN
Hydrogels made with semi-interpenetrating networks of the oligomerized polyphenol tannic acid, and poly(acrylamide), exhibit high stiffness and toughness. However, the structure property relationships that give rise to enhanced mechanical properties is not well understood. Herein, we systematically investigate the hydrogels using small angle X-ray scattering and small and Ultra-small angle neutron scattering within a wide length scale range (1 nm to 20 µm), polarized optical microscopy, and rheology. Small angle X-ray and neutron scattering reveal the presence of micron sized hydrogen bonded clusters in the hydrogels. Breaking of hydrogen bonded clusters above a critical solution temperature was clearly observed in the small angle neutron scattering data. Polarized optical microscopy show enhanced anisotropy for the gels with oligomerized tannic acid incorporated - when compared to gels with monomeric tannic acid. Rheological studies at varying temperatures nicely corroborate the structural changes observed at high temperatures and reveal a self-healing behavior of the gels. The knowledge gained from this study will aid in rational design of hydrogels for biomedical applications.
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Hidrogeles , Taninos , Hidrogeles/química , Enlace de Hidrógeno , Dispersión del Ángulo Pequeño , Reología , HidrógenoRESUMEN
A combination of chemotherapy and chemodynamic therapy (CDT) is being developed to improve the theranostic efficacy and biological safety of current therapies. However, most CDT agents are restricted due to complex issues such as multiple components, low colloidal stability, carrier-associated toxicity, insufficient reactive oxygen species generation, and poor targeting efficacy. To overcome these problems, a novel nanoplatform composed of fucoidan (Fu) and iron oxide (IO) nanoparticles (NPs) was developed to achieve chemotherapy combined with CDT synergistic treatment with a facile self-assembling manner, and the NPs were made up of Fu and IO, in which the Fu was not only used as a potential chemotherapeutic but was also designed to stabilize the IO and target P-selectin-overexpressing lung cancer cells, thereby producing oxidative stress and thus synergizing the CDT efficacy. The Fu-IO NPs exhibited a suitable diameter below 300 nm, which favored their cellular uptake by cancer cells. Microscopic and MRI data confirmed the lung cancer cellular uptake of the NPs due to active Fu targeting. Moreover, Fu-IO NPs induced efficient apoptosis of lung cancer cells, and thus offer significant anti-cancer functions by potential chemotherapeutic-CDT.
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Neoplasias Pulmonares , Nanopartículas , Neoplasias , Humanos , Medicina de Precisión , Selectina-P , Línea Celular Tumoral , Nanomedicina Teranóstica , Neoplasias/tratamiento farmacológico , Estrés Oxidativo , Neoplasias Pulmonares/tratamiento farmacológico , Nanopartículas Magnéticas de Óxido de Hierro , Nanopartículas/uso terapéuticoRESUMEN
Traditional thrombolytic therapeutics for vascular blockage are affected by their limited penetration into thrombi, associated off-target side effects, and low bioavailability, leading to insufficient thrombolytic efficacy. It is hypothesized that these limitations can be overcome by the precisely controlled and targeted delivery of thrombolytic therapeutics. A theranostic platform is developed that is biocompatible, fluorescent, magnetic, and well-characterized, with multiple targeting modes. This multimodal theranostic system can be remotely visualized and magnetically guided toward thrombi, noninvasively irradiated by near-infrared (NIR) phototherapies, and remotely activated by actuated magnets for additional mechanical therapy. Magnetic guidance can also improve the penetration of nanomedicines into thrombi. In a mouse model of thrombosis, the thrombosis residues are reduced by ≈80% and with no risk of side effects or of secondary embolization. This strategy not only enables the progression of thrombolysis but also accelerates the lysis rate, thereby facilitating its prospective use in time-critical thrombolytic treatment.
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Terapia Trombolítica , Trombosis , Ratones , Animales , Medicina de Precisión , Fibrinolíticos/química , Fibrinolíticos/uso terapéutico , Trombosis/diagnóstico por imagen , Trombosis/tratamiento farmacológico , Fenómenos MagnéticosRESUMEN
The solution state structure of κ-carrageenan is typically described as a 'random coil', to indicate a lack of defined secondary structure elements. From this starting point the assignment of an optical-rotation-detected change that follows the introduction of particular ions to such solutions as a 'coil-to-helix transition' seems unambiguous, and thus the canonical description of this important biopolymer's gelling behaviour was born. However, the notion that κ-carrageenan exists in solution as a random coil, devoid of secondary structure, has been questioned a number of times previously in the literature, particularly by the molecular modelling and NMR communities. Regrettably, there has been little desire to-date to address these largely overlooked studies or consider their implications for the nature of the so-called 'coil-to-helix transition'. Despite evidence to the contrary, the random-coil-paradigm has prevailed. Here, new data from synchrotron-enabled solution-state x-ray scattering experiments, combined with state-of-the-art atomistic molecular dynamics simulations, are used to show that the solution-state structure of κ-carrageenan in fact retains many of the helical motifs present in the solid-state, as inferred from fibre diffraction data. Furthermore, no evidence is found to suggest that single chains undergo any uni-molecular conformational transition upon the addition of ions. These findings once again challenge the paradigm that κ-carrageenan exists as a 'random coil' in the solution state, and thereby question the long held assumption that a uni-molecular 'coil-to-helix transition' precedes the dimerization of helices.
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Simulación de Dinámica Molecular , Carragenina/química , Iones , Estructura Secundaria de Proteína , Rayos XRESUMEN
DNA-templated metallization is broadly investigated in the fabrication of metallic structures by virtue of the unique DNA-metal ion interaction. However, current DNA-templated synthesis is primarily carried out based on pure DNA in an aqueous solution. In this study, we present in situ synthesis of metallic structures in a natural DNA complex bulk film by UV light irradiation, where the growth of silver particles is resolved by in situ time-resolved small-angle X-ray scattering and dielectric spectroscopy. Our studies provide physical insights into the kinetics and mechanisms of natural DNA metallization, in correlation with the multi-stage switching operations in the bulk phase, paving the way towards the development of versatile biomaterial composites with tunable physical properties for optical storage, plasmonics, and catalytic applications.
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Functional bionanocomposites have evoked immense research interests in many fields including biomedicine, food packaging, and environmental applications. Supramolecular self-assembled bionanocomposite materials fabricated by biopolymers and two-dimensional (2D) nanomaterials have particularly emerged as a compelling material due to their biodegradable nature, hierarchical structures, and designable multifunctions. However, construction of these materials with tunable properties has been still challenging. Here, we report a self-assembled, flexible, and antioxidative collagen nanocomposite film (CNF) via regulating supramolecular interactions of type I collagen and tannic acid (TA)-functionalized 2D synthetic clay nanoplatelets Laponite (LAP). Specifically, TA-coordinated LAP (LAP-TA) complexes were obtained via chelation and hydrogen bonding between TA and LAP clay nanoplatelets and further used to stabilize the triple-helical confirmation and fibrillar structure of the collagen via hydrogen bonding and electrostatic interactions, forming a hierarchical microstructure. The obtained transparent CNF not only exhibited the reinforced thermal stability, enzymatic resistance, tensile strength, and hydrophobicity but also good water vapor permeability and antioxidation. For example, the tensile strength was improved by over 2000%, and the antioxidant property was improved by 71%. Together with the simple fabrication process, we envision that the resulting CNF provides greater opportunities for versatile bionanocomposites design and fabrication serving as a promising candidate for emerging applications, especially food packaging and smart wearable devices.
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Nanocompuestos , Polifenoles , Arcilla , Colágeno , Embalaje de Alimentos , Nanocompuestos/químicaRESUMEN
The innate cartilage extracellular matrix is avascular and plays a vital role in innate chondrocytes. Recapping the crucial components of the extracellular matrix in engineered organs via polymeric gels and bioinspired approaches is promising for improving the regenerative aptitude of encapsulated cartilage/chondrocytes. Conventional gel formation techniques for polymeric materials rely on employing oxidative crosslinking, which is constrained in this avascular environment. Further, poor mechanical properties limit the practical applications of polymeric gels and reduce their therapeutic efficacy. Herein, the purpose of this study was to develop a bioadhesive gel possessing dual crosslinking for engineering cartilage. Tyramine (TYR) was first chemically conjugated to the alginate (ALG) backbone to form an ALG-TYR precursor, followed by the addition of calcium peroxide (CaO2); calcium ions of CaO2 physically crosslink with ALG, and oxygen atoms of CaO2 chemically crosslink TYR with tyrosinase, thus enabling dual/enhanced crosslinking and possessing injectability. The ALG-TYR/tyrosinase/CaO2 gel system was chemically, mechanically, cellularly, and microscopically characterized. The gel system developed herein was biocompatible and showed augmented mechanical strength. The results showed, for the first time, that CaO2 supplementation preserved cell viability and enhanced the crosslinking ability, bioadhesion, mechanical strength, chondrogenesis, and stability for cartilage regeneration.
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Alginatos , Monofenol Monooxigenasa , Alginatos/química , Cartílago , Condrocitos , Condrogénesis , Hidrogeles/química , Peróxidos , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , TiraminaRESUMEN
Gene therapy is one of the most important developments for modern medicine. As such methods for the compaction and delivery of nucleic acids bearing therapeutic sequences is essential. The quest for non-viral carriers of nucleic acids has produced a number of possible candidates with dendrimer being among the most promising. Their hyper-branched structure and well-defined size together with low cytotoxicity has found success in both ex-vivo and in-vivo studies. The compaction of DNA with dendrimer has produced a rich array of different structures depending on the physiochemical conditions. Mechanisms that drive the compaction have been shown to be a number of physical interactions that reduce the large polymeric entity from 100s of nanometers to some tens of nanometers to fit into the cell nucleus. The mechanisms driving the compaction of DNA will be discussed in detail while the focus will be directed to tuning the structural properties of the complexes and their structural characterization using small-angle scattering techniques.