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Dentin hypersensitivity is an oral health concern affecting a large percentage of the world's adult population. Occlusion of the exposed dentinal tubules is among the treatment options available, and silver diammine fluoride (SDF) is an occluding agent used for interrupting or dampening the stimulus of the dental pulp nerves that produce pain. In addition to dentin permeability testing, the evaluation of desensitizing agents occluding dentinal tubules strongly relies on microscopic techniques, such as scanning electron microscopy (SEM). Limitations of SEM are that it provides only surface images that lack detailed information on the depth of penetration and amount of material present within the treated specimen, and it is prone to sample preparation artifacts. Here, we present high-resolution X-ray computed tomography (nano-CT) as a potential method for investigating dentin specimens with occluded tubules. We studied human dentin treated with SDF as an exemplary dentinal occlusion treatment option. We evaluated the silver deposits formed on the dentin surface region near the dentinal tubules and in the tubular regions using cross-section SEM, Energy Dispersive X-ray (EDX) analysis, and nano-CT. The resulting images obtained by SEM and nano-CT had comparable resolutions, and both techniques produced images of the tubules' occlusion. Nano-CT provided three-dimensional images adequate to quantitate tubule size and orientation in space. Moreover, it enabled clear visualization of dentinal tubules in any virtual plane and estimation of the amount and depth of occluding material. Thus, nano-CT has the potential to be a valuable technique for evaluating the occluding effects of virtually any material applied to dentinal tubules, supporting deciding between the best occluding treatment options.
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Artefactos , Tomografía Computarizada por Rayos X , Adulto , Humanos , Microscopía Electrónica de Rastreo , Dentina/diagnóstico por imagenRESUMEN
Delivery of growth factors (GFs) is challenging for regulation of cell proliferation and differentiation due to their rapid inactivation under physiological conditions. Here, a bioactive polyelectrolyte multilayer (PEM) is engineered by the combination of thermoresponsive poly(N-isopropylacrylamide) (PNIPAM) and glycosaminoglycans to be used as reservoir for GF storage. PNIPAM-grafted-chitosan (PChi) with two degrees of substitution (DS) are synthesized, namely LMW* (DS 0.14) and HMW (DS 0.03), by grafting low (2 kDa) and high (10 kDa) molecular weight of PNIPAM on the backbone of chitosan (Chi) to be employed as polycations to form PEM with the polyanion heparin (Hep) at pH 4. Subsequently, PEMs are chemically crosslinked to improve their stability at physiological pH 7.4. Resulting surface and mechanical properties indicate that PEM containing HMW is responsive to temperature at 20 °C and 37 °C, while LMW is not. More importantly, Hep as terminal layer combined with HMW allows not only a better retention of the adhesive protein vitronectin but also a sustained release of FGF-2 at 37 °C. With the synergistic effect of vitronectin and matrix-bound FGF-2, significant promotion on adhesion, proliferation, and migration of 3T3 mouse embryonic fibroblasts is achieved on HMW-containing PEM compared to Chi-containing PEM and exogenously added FGF-2. Thus, PEM containing PNIPAM in combination with bioactive glycosaminoglycans like Hep represents a versatile approach to fabricate a GF delivery system for efficient cell culture, which can be potentially served as cell culture substrate for production of (stem) cells and bioactive wound dressing for tissue regeneration.
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Quitosano , Heparina , Animales , Ratones , Heparina/farmacología , Heparina/química , Quitosano/química , Quitosano/farmacología , Factor 2 de Crecimiento de Fibroblastos/farmacología , Vitronectina/farmacología , Adhesión Celular , Fibroblastos , Glicosaminoglicanos/química , Glicosaminoglicanos/farmacologíaRESUMEN
Elastin is an essential extracellular matrix protein that enables tissues and organs such as arteries, lungs, and skin, which undergo continuous deformation, to stretch and recoil. Here, an approach to fabricating artificial elastin with close-to-native molecular and mechanical characteristics is described. Recombinantly produced tropoelastin are polymerized through coacervation and allysine-mediated cross-linking induced by pyrroloquinoline quinone (PQQ). A technique that allows the recovery and repeated use of PQQ for protein cross-linking by covalent attachment to magnetic Sepharose beads is developed. The produced material closely resembles natural elastin in its molecular, biochemical, and mechanical properties, enabled by the occurrence of the cross-linking amino acids desmosine, isodesmosine, and merodesmosine. It possesses elevated resistance against tryptic proteolysis, and its Young's modulus ranging between 1 and 2 MPa is similar to that of natural elastin. The approach described herein enables the engineering of mechanically resilient, elastin-like materials for biomedical applications.
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Elastina , Tropoelastina , Elastina/química , Tropoelastina/química , Aminoácidos , ProteolisisRESUMEN
Perioperative hypothermia is still common and has relevant complication for the patient. An effective perioperative thermal management requires essentially an accurate method to measure core temperature. So far, only one study has investigated the new Temple Touch Pro™ (Medisim Ltd., Beit-Shemesh, Israel). during anesthesia Therefore, we assessed the agreement between the Temple Touch Pro™ thermometer (TTP) and distal esophageal temperature (TEso) in a second study. After approval by the local ethics committee we studied 100 adult patients undergoing surgery with general anesthesia. Before induction of anesthesia the TTP sensor unit was attached to the skin above the temporal artery. After induction of anesthesia an esophageal temperature probe was placed in the distal esophagus. Recordings started 10 min after placement of the esophageal temperature probe to allow adequate warming of the probes. Pairs of temperature values were documented in five-minute intervals until emergence of anesthesia. Accuracy of the two methods was assessed by Bland-Altman comparisons of differences with multiple measurements. Core temperatures obtained with the TTP in adults showed a mean bias of -0.04 °C with 95% limits of agreement within - 0.99 °C to + 0.91 °C compared to an esophageal temperature probe. We consider the TTP as a reasonable tool for perioperative temperature monitoring. It is not accurate enough to be used as a reference method in scientific studies, but may be a useful tool especially for conscious patients undergoing neuraxial anesthesia or regional anesthesia with sedation. Trial registration This study was registered in the German Clinical Trials Register (DRKS-ID: 00024050), day of registration 12/01/2021.
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Hipotermia , Tacto , Adulto , Humanos , Temperatura , Temperatura Corporal , Anestesia GeneralRESUMEN
A gene-activated surface coating is presented as a strategy to design smart biomaterials for bone tissue engineering. The thin-film coating is based on polyelectrolyte multilayers composed of collagen I and chondroitin sulfate, two main biopolymers of the bone extracellular matrix, which are fabricated by layer-by-layer assembly. For further functionalization, DNA/lipid-nanoparticles (lipoplexes) are incorporated into the multilayers. The polyelectrolyte multilayer fabrication and lipoplex deposition are analyzed by surface sensitive analytical methods that demonstrate successful thin-film formation, fibrillar structuring of collagen, and homogenous embedding of lipoplexes. Culture of mesenchymal stem cells on the lipoplex functionalized multilayer results in excellent attachment and growth of them, and also, their ability to take up cargo like fluorescence-labelled DNA from lipoplexes. The functionalization of the multilayer with lipoplexes encapsulating DNA encoding for transient expression of bone morphogenetic protein 2 induces osteogenic differentiation of mesenchymal stem cells, which is shown by mRNA quantification for osteogenic genes and histochemical staining. In summary, the novel gene-functionalized and extracellular matrix mimicking multilayer composed of collagen I, chondroitin sulfate, and lipoplexes, represents a smart surface functionalization that holds great promise for tissue engineering constructs and implant coatings to promote regeneration of bone and other tissues.
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Sulfatos de Condroitina , Osteogénesis , Polielectrolitos , Diferenciación Celular , Colágeno , Colágeno Tipo I/genética , Técnicas de Transferencia de Gen , ADN/metabolismo , Matriz Extracelular/metabolismoRESUMEN
The aim of the current paper is to summarize the results of the International CytoSorb Registry. Data were collected on patients of the intensive care unit. The primary endpoint was actual in-hospital mortality compared to the mortality predicted by APACHE II score. The main secondary endpoints were SOFA scores, inflammatory biomarkers and overall evaluation of the general condition. 1434 patients were enrolled. Indications for hemoadsorption were sepsis/septic shock (N = 936); cardiac surgery perioperatively (N = 172); cardiac surgery postoperatively (N = 67) and "other" reasons (N = 259). APACHE-II-predicted mortality was 62.0±24.8%, whereas observed hospital mortality was 50.1%. Overall SOFA scores did not change but cardiovascular and pulmonary SOFA scores decreased by 0.4 [-0.5;-0.3] and -0.2 [-0.3;-0.2] points, respectively. Serum procalcitonin and C-reactive protein levels showed significant reduction: -15.4 [-19.6;-11.17] ng/mL; -17,52 [-70;44] mg/L, respectively. In the septic cohort PCT and IL-6 also showed significant reduction: -18.2 [-23.6;-12.8] ng/mL; -2.6 [-3.0;-2.2] pg/mL, respectively. Evaluation of the overall effect: minimal improvement (22%), much improvement (22%) and very much improvement (10%), no change observed (30%) and deterioration (4%). There was no significant difference in the primary outcome of mortality, but there were improvements in cardiovascular and pulmonary SOFA scores and a reduction in PCT, CRP and IL-6 levels. Trial registration: ClinicalTrials.gov Identifier: NCT02312024 (retrospectively registered).
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Sepsis , Choque Séptico , Humanos , Enfermedad Crítica/terapia , Polipéptido alfa Relacionado con Calcitonina , Proteína C-Reactiva , Interleucina-6 , Sepsis/terapia , Sepsis/metabolismo , Curva ROC , Pronóstico , Biomarcadores , Sistema de RegistrosRESUMEN
Metal ions are important effectors of protein and cell functions. Here, polyelectrolyte multilayers (PEMs) made of chitosan (Chi) and alginate (Alg) were doped with different metal ions (Ca2+, Co2+, Cu2+, and Fe3+), which can form bonds with their functional groups. Ca2+ and Fe3+ ions can be deposited in PEM at higher quantities resulting in more positive ζ potentials and also higher water contact angles in the case of Fe3+. An interesting finding was that the exposure of PEM to metal ions decreases the elastic modulus of PEM. Fourier transformed infrared (FTIR) spectroscopy of multilayers provides evidence of interaction of metal ions with the carboxylic groups of Alg but not for hydroxyl and amino groups. The observed changes in wetting and surface potential are partly related to the increased adhesion and proliferation of multipotent C3H10T1/2 fibroblasts in contrast to plain nonadhesive [Chi/Alg] multilayers. Specifically, PEMs doped with Cu2+ and Fe3+ ions greatly promote cell attachment and adipogenic differentiation, which indicates that changes in not only surface properties but also the bioactivity of metal ions play an important role. In conclusion, metal ion-doped multilayer coatings made of alginate and chitosan can promote the differentiation of multipotent cells on implants without the use of other morphogens like growth factors.
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Alginatos , Quitosano , Adipogénesis , Alginatos/química , Alginatos/farmacología , Quitosano/farmacología , Iones , Polielectrolitos/química , Polielectrolitos/farmacología , Células Madre , Agua/químicaRESUMEN
The sensory end-organs responsible for hearing and balance in the mammalian inner ear are connected via a small membranous duct known as the ductus reuniens (also known as the reuniting duct (DR)). The DR serves as a vital nexus linking the hearing and balance systems by providing the only endolymphatic connection between the cochlea and vestibular labyrinth. Recent studies have hypothesized new roles of the DR in inner ear function and disease, but a lack of knowledge regarding its 3D morphology and spatial configuration precludes testing of such hypotheses. We reconstructed the 3D morphology of the DR and surrounding anatomy using osmium tetroxide micro-computed tomography and digital visualizations of three human inner ear specimens. This provides a detailed, quantitative description of the DR's morphology, spatial relationships to surrounding structures, and an estimation of its orientation relative to head position. Univariate measurements of the DR, inner ear, and cranial planes were taken using the software packages 3D Slicer and Zbrush. The DR forms a narrow, curved, flattened tube varying in lumen size, shape, and wall thickness, with its middle third being the narrowest. The DR runs in a shallow bony sulcus superior to the osseus spiral lamina and adjacent to a ridge of bone that we term the "crista reuniens" oriented posteromedially within the cranium. The DR's morphology and structural configuration relative to surrounding anatomy has important implications for understanding aspects of inner ear function and disease, particularly after surgical alteration of the labyrinth and potential causative factors for Ménière's disease.
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Vestíbulo del Laberinto , Humanos , Audición , Enfermedad de Meniere/diagnóstico por imagen , Vestíbulo del Laberinto/anatomía & histología , Vestíbulo del Laberinto/diagnóstico por imagen , Microtomografía por Rayos XRESUMEN
OBJECTIVE: Silver diamine fluoride (SDF) is effective in treatment of dentin hypersensitivity and caries lesions. However, the non-viscous solution does not easily allow clinicians to control the application area. A 38% SDF experiment gel was compared in vitro to commercial SDF for its ability to penetrate and occlude dentinal tubules. MATERIALS AND METHODS: Human root surface dentin specimens were treated with gelled or standard 38% SDF or negative control. Penetration behavior was established by Drop Shape Analysis. Precipitates at the surface and within tubules were analyzed by SEM and EDX after treatment; Results: penetration depths up to 500 µm were observed for both SDF formulations. Both formulations occluded dentinal tubules similarly. Precipitates on the dentin surface and within dentinal tubules were found for both SDF formulations, with a slight tendency for the experimental gel SDF product to be more abundant than the commercially available one. DISCUSSION: behavior of the experimental 38% SDF gel formulation appeared indistinguishable from the commercial 38% SDF product with respect to dentinal tubule penetration and occlusion. CONCLUSIONS: The experimental 38% SDF gel may be a suitable intervention for the prevention of dentin hypersensitivity.
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The effective delivery of drugs to the inner ear is still an unmet medical need. Local controlled drug delivery to this sensory organ is challenging due to its location in the petrous bone, small volume, tight barriers, and high vulnerability. Local intracochlear delivery of drugs would overcome the limitations of intratympanic (extracochlear) and systemic drug application. The requirements for such a delivery system include small size, appropriate flexibility, and biodegradability. We have developed biodegradable PLGA-based implants for controlled intracochlear drug release that can also be used in combination with cochlear implants (CIs), which are implantable neurosensory prosthesis for hearing rehabilitation. The drug carrier system was tested for implantation in the human inner ear in 11 human temporal bones. In five of the temporal bones, CI arrays from different manufacturers were implanted before insertion of the biodegradable PLGA implants. The drug carrier system and CI arrays were implanted into the scala tympani through the round window. Implanted temporal bones were evaluated by ultra-high-resolution computed tomography (µ-CT) to illustrate the position of implanted electrode carriers and the drug carrier system. The µ-CT measurements revealed the feasibility of implanting the PLGA implants into the scala tympani of the human inner ear and co-administration of the biodegradable PLGA implant with a CI array.
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Implantación Coclear , Implantes Cocleares , Oído Interno , Humanos , Preparaciones Farmacéuticas , Hueso Temporal/diagnóstico por imagen , Hueso Temporal/cirugíaRESUMEN
The stratum corneum (SC) is the largest physical barrier of the human body. It protects against physical, chemical and biological damages, and avoids evaporation of water from the deepest skin layers. For its correct functioning, the homeostasis of the SC lipid matrix is fundamental. An alteration of the lipid matrix composition and in particular of its ceramide (CER) fraction can lead to the development of pathologies such as atopic dermatitis and psoriasis. Different studies showed that the direct replenishment of SC lipids on damaged skin had positive effects on the recovery of its barrier properties. In this work, cerosomes, i.e. liposomes composed of SC lipids, have been successfully prepared in order to investigate the mechanism of interaction with a model SC lipid matrix. The cerosomes contain CER[NP], D-CER[AP], stearic acid and cholesterol. In addition, hydrogenated soybean phospholipids have been added to one of the formulations leading to an increased stability at neutral pH. For the mode of action studies, monolayer models at the air-water interface and on solid support have been deployed. The results indicated that a strong interaction occurred between SC monolayers and the cerosomes. Since both systems were negatively charged, the driving force for the interaction must be based on the ability of CERs head groups to establish intermolecular hydrogen bonding networks that energetically prevailed against the electrostatic repulsion. This work proved for the first time the mode of action by which cerosomes exploit their function as skin barrier repairing agents on the SC.
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Polyelectrolyte multilayers (PEMs) consisting of the polysaccharides hyaluronic acid (HA) as the polyanion and chitosan (Chi) as the polycation were prepared with layer-by-layer technique (LbL). The [Chi/HA]5 multilayers were exposed to solutions of metal ions (Ca2+, Co2+, Cu2+ and Fe3+). Binding of metal ions to [Chi/HA]5 multilayers by the formation of complexes with functional groups of polysaccharides modulates their physical properties and the bioactivity of PEMs with regard to the adhesion and function of multipotent murine C3H10T1/2 embryonic fibroblasts. Characterization of multilayer formation and surface properties using different analytical methods demonstrates changes in the wetting, surface potential and mechanical properties of multilayers depending on the concentration and type of metal ion. Most interestingly, it is observed that Fe3+ metal ions greatly promote adhesion and spreading of C3H10T1/2 cells on the low adhesive [Chi/HA]5 PEM system. The application of intermediate concentrations of Cu2+, Ca2+ and Co2+ as well as low concentrations of Fe3+ to PEMs also results in increased cell spreading. Moreover, it can be shown that complex formation of PEMs with Cu2+ and Fe3+ ions leads to increased metabolic activity in cells after 24 h and induces cell differentiation towards adipocytes in the absence of any additional adipogenic media supplements. Overall, complex formation of [Chi/HA]5 PEM with metal ions like Cu2+ and Fe3+ represents an interesting and cheap alternative to the use of growth factors for making cell-adhesive coatings and guiding stem cell differentiation on implants and scaffolds to regenerate connective-type of tissues.
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Quitosano , Ácido Hialurónico , Animales , Adhesión Celular , Diferenciación Celular , Fibroblastos , Iones , Ratones , Propiedades de SuperficieRESUMEN
Endothelin-1 (ET-1) is a member of the endothelin family of peptide hormones first discovered as endothelium-derived mediators regulating vascular tone. ET-1 also regulates the proliferation and differentiation of bone cells that synthesize fibroblast growth factor 23 (FGF23). FGF23 is a hormone controlling renal phosphate and vitamin D metabolism. Here, we studied the role of ET-1 and endothelin receptor B (ETB) for FGF23 production. Fgf23 gene expression was studied in IDG-SW3 bone cells by quantitative RT-PCR. ETB-expressing (etb+/+ ) and rescued ETB-deficient mice (etb-/- ) were studied in metabolic cages. Their serum FGF23, PTH, and 1,25(OH)2 D3 concentrations were determined by ELISA, serum and urinary phosphate and Ca2+ by photometric methods. ET-1 and ETB agonist sarafotoxin 6c suppressed Fgf23 mRNA in IDG-SW3 cells. Serum C-terminal and intact FGF23 as well as bone Fgf23 mRNA levels were significantly higher in etb-/- mice than in etb+/+ mice. Renal phosphate excretion was significantly higher in etb-/- mice despite lower phosphate levels. In addition, etb-/- animals exhibited calciuria and a significantly higher serum 1,25(OH)2 D3 concentration compared to etb+/+ mice. In conclusion, ETB-dependent ET-1 signaling is a potent suppressor of FGF23 formation. This effect is likely to be of clinical relevance given the use of endothelin receptor antagonists in various diseases.
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Factores de Crecimiento de Fibroblastos/metabolismo , Regulación de la Expresión Génica , Receptor de Endotelina B/fisiología , Animales , Huesos/metabolismo , Calcio/metabolismo , Femenino , Factor-23 de Crecimiento de Fibroblastos , Factores de Crecimiento de Fibroblastos/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Fosfatos/metabolismoRESUMEN
BACKGROUND: Prevention of inadvertent hypothermia is recommended for procedures >30 minutes because hypothermia increases the risk of myocardial ischemia, intraoperative blood loss, transfusion and wound complications. Therefore, short warming interruptions between pre-warming and intraoperative warming might result in lower hypothermia rates. The aim of this retrospective investigation was to determine whether the incidence of inadvertent intraoperative hypothermia was affected by the warming interruption. METHODS: The lowest intraoperative body core temperature value and the warming interruption time were taken from anaesthesia records. Body core temperature was recorded continuously, and a patient was classified to be hypothermic if the lowest recorded temperature value was <36°C. Hypothermia rates and the correlation between warming interruption times and intraoperative hypothermia rates were calculated. RESULTS: Five thousand eighty-four patients were analysed. The intraoperative hypothermia rate was 15.3%. Nineteen patients (0.4%) had a recorded temperature of <35.0°C. An increase in forced-air warming interruption time was significantly associated with an increase in intraoperative hypothermia rates (P < .0001). Patients with interruptions in forced-air warming >20 minutes showed significantly higher hypothermia rates than those with interruptions of ≤20 minutes (P < .0001). CONCLUSION: Intraoperative hypothermia rates increased significantly with longer forced-air warming interruptions between pre-warming and intraoperative warming. Short warming interruptions can preserve the effect of pre-warming and are associated with low intraoperative hypothermia rates.
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Hipotermia/prevención & control , Cuidados Intraoperatorios/métodos , Complicaciones Intraoperatorias/prevención & control , Recalentamiento/métodos , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , TiempoRESUMEN
To avoid pathogen-associated molecular pattern recognition, the hemibiotrophic maize pathogen Colletotrichum graminicola secretes proteins mediating the establishment of biotrophy. Targeted deletion of 26 individual candidate genes and seven gene clusters comprising 32 genes of C. graminicola identified a pathogenicity cluster (CLU5) of five co-linear genes, all of which, with the exception of CLU5b, encode secreted proteins. Targeted deletion of all genes of CLU5 revealed that CLU5a and CLU5d are required for full appressorial penetration competence, with virulence deficiencies independent of the host genotype and organ inoculated. Cytorrhysis experiments and microscopy showed that Δclu5a mutants form pressurized appressoria, but they are hampered in forming penetration pores and fail to differentiate a penetration peg. Whereas Δclu5d mutants elicited WT-like papillae, albeit at increased frequencies, papillae induced by Δclu5a mutants were much smaller than those elicited by the WT. Synteny of CLU5 is not only conserved in Colletotrichum spp. but also in additional species of Sordariomycetes including insect pathogens and saprophytes suggesting importance of CLU5 for fungal biology. Since CLU5a and CLU5d also occur in non-pathogenic fungi and since they are expressed prior to plant invasion and even in vegetative hyphae, the encoded proteins probably do not act primarily as effectors.
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Colletotrichum/metabolismo , Proteínas Fúngicas/metabolismo , Enfermedades de las Plantas/microbiología , Zea mays/microbiología , Colletotrichum/genética , Colletotrichum/patogenicidad , Proteínas Fúngicas/genética , Hifa/genética , Hifa/metabolismo , Hifa/patogenicidad , Familia de Multigenes , Eliminación de Secuencia , VirulenciaRESUMEN
In this study, the morphology of an immiscible polymer blend system at various regions of interests was analyzed using different microanalytical methods with varying surface sensitivities. As a model immiscible polymer blend, a HDPE/PP (80/20â¯wt%) polymer film was used. The blend film was subjected to polarized light microscopy (PLM), scanning electron microscopy (SEM), atomic force microscopy (AFM), transmission electron microscopy (TEM) and time of flight secondary ion mass spectrometry (ToF-SIMS). The obtained results were compared regarding the sensitivities, informational values and overall applicability of the analytical methods. It was evaluated which methods can be applied for a fast analysis of the morphology (surface and bulk) of the immiscible polymer blend with low preparation efforts, which is especially important for the analysis of new materials, for example materials manufactured via recycling. It was demonstrated that PLM, as well as SEM on wet-etched material, provide sufficient information to evaluate the bulk morphology. Additionally, the presented study shows the advantage of applying ToF-SIMS imaging for the characterization of the surface of immiscible polymer blend. As expected, the domain distribution of HDPE and PP varied between the bulk and the surface of the films. The proposed procedures can be taken as a guideline for other investigations concerning the morphology of heterogeneous polyolefin systems.
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Advanced technologies for controlled cell adhesion and detachment in novel biointerface designs profit from stimuli-responsive systems that are able to react to their environment. Here, a multilayer system made of thiolated chitosan and thiolated chondroitin sulfate was constructed, with the potential of switchable inter- and intramolecular thiol/disulfide interactions representing a redox-sensitive nanoplatform. Owing to the formation and cleavage of inherent disulfide bonds by oxidation and reduction, surface properties of the multilayer can be controlled toward protein adsorption/desorption and cell adhesion in a reversible manner. Oxidation of thiols by chloramine-T promotes fibronectin (FN) adsorption and fibroblast cell adhesion, whereas the reduction by tris(2-carboxyethyl)phosphine reverses these effects, leading to low FN adsorption and little cell adhesion and spreading. These effects on the biological systems are related to significant changes of wetting properties, zeta potential, and mechanical properties of these multilayer films. The system presented may be useful for biomedical applications as responsive and obedient surfaces in medical implants and support tissue regeneration.
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Adhesión Celular/efectos de los fármacos , Quitosano/química , Sulfatos de Condroitina/química , Fibroblastos/efectos de los fármacos , Adsorción , Células Cultivadas , Cloraminas/metabolismo , Fibroblastos/metabolismo , Fibronectinas/metabolismo , Humanos , Oxidación-Reducción , Compuestos de Sulfhidrilo/química , Propiedades de Superficie , Compuestos de Tosilo/metabolismoRESUMEN
BACKGROUND: Cells possess a specialized machinery through which they can sense physical as well as chemical alterations in their surrounding microenvironment that affect their cellular behavior. AIM: In this study, we aim to establish a polyelectrolyte multilayer system of 24 layers of poly-l-lysine and hyaluronic acid to control stem cell response after chemical cross-linking. METHODS AND RESULTS: The multilayer build-up process is monitored using different methods, which show that the studied polyelectrolyte multilayer system grows exponentially following the islands and islets theory. Successful chemical cross-linking is monitored by an increased zeta potential toward negative magnitude and an extraordinary growth in thickness. Human adipose-derived stem cells are used here and a relationship between cross-linking degree and cell spreading is shown as cells seeded on higher cross-linked polyelectrolyte multilayer show enhanced spreading. Furthermore, cells that fail to establish focal adhesions on native and low cross-linked polyelectrolyte multilayer films do not proliferate to a high extent in comparison to cells seeded on highly cross-linked polyelectrolyte multilayer, which also show an increased metabolic activity. Moreover, this study shows the relation between cross-linking degree and human adipose-derived stem cell lineage commitment. Histological staining reveals that highly cross-linked polyelectrolyte multilayers support osteogenic differentiation, whereas less cross-linked and native polyelectrolyte multilayers support adipogenic differentiation in the absence of any specific inducers. CONCLUSION: Owing to the precise control of polyelectrolyte multilayer properties such as potential, wettability, and viscoelasticity, the system presented here offers great potential for guided stem cell differentiation in regenerative medicine, especially in combination with materials exhibiting a defined surface topography.
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Reactivos de Enlaces Cruzados/química , Ácido Hialurónico/química , Células Madre Mesenquimatosas/citología , Polilisina/química , Diferenciación Celular , Reactivos de Enlaces Cruzados/farmacología , HumanosRESUMEN
Control of the biomaterial properties through stimuli-responsive polymeric platforms has become an essential technique in recent biomedical applications. A multilayer system of thiolated chitosan (t-Chi) and thiolated chondroitin sulfate (t-CS), consisting of five double layers ([t-Chi/t-CS]5), was fabricated here by applying a layer-by-layer coating strategy. To represent a novel class of chemically tunable nanostructures, the ability to cross-link pendant thiol groups was tested by a rise from pH 4 during layer formation to pH 9.3 and a more powerful chemical stimulus by using chloramine-T (ChT). Following both treatments, the resulting multilayers showed stimuli-dependent behavior, as demonstrated by their content of free thiols, wettability, surface charge, elastic modulus, roughness, topography, thickness, and binding of fibronectin. Studies with human dermal fibroblasts further demonstrated the favorable potential of the ChT-responsive multilayers as a cell-adhesive surface compared to pH-induced cross-linking. Because the [t-Chi/t-CS]5 multilayer system is responsive to stimuli such as the pH and redox environment, multilayer systems with disulfide bond formation may help to tailor their interaction with cells, film degradation, and controlled release of bioactive substances like growth factors in a stimuli-responsive manner useful in future wound healing and tissue engineering applications.