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Injectable bone substitutes (IBSs) represent a compelling choice for bone tissue regeneration, as they can be exploited to optimally fill complex bone defects in a minimally invasive manner. In this context, in situ gelling methylcellulose (MC) hydrogels may be engineered to be free-flowing injectable solutions at room temperature and gels upon exposure to body temperature. Moreover, incorporating a suitable inorganic phase can further enhance the mechanical properties of MC hydrogels and promote mineralization, thus assisting early cell adhesion to the hydrogel and effectively guiding bone tissue regeneration. In this work, thermo-responsive IBSs were designed selecting MC as the organic matrix and calcium phosphate (CaP) or CaP modified with graphene oxide (CaPGO) as the inorganic component. The resulting biocomposites displayed a transition temperature around body temperature, preserved injectability even after loading with the inorganic components, and exhibited adequate retention on an ex vivo calf femoral bone defect model. The addition of CaP and CaPGO promoted the in vitro mineralization process already 14 days after immersion in simulated body fluid. Interestingly, combined X-ray diffraction and solid state nuclear magnetic resonance characterizations revealed that the formed biomimetic phase was constituted by crystalline hydroxyapatite and amorphous calcium phosphate. In vitro biological characterization revealed the beneficial impact of CaP and CaPGO, indicating their potential in promoting cell adhesion, proliferation and osteogenic differentiation. Remarkably, the addition of GO, which is very attractive for its bioactive properties, did not negatively affect the injectability of the hydrogel nor the mineralization process, but had a positive impact on cell growth and osteogenic differentiation on both pre-differentiated and undifferentiated cells. Overall, the proposed formulations represent potential candidates for use as IBSs for application in bone regeneration both under physiological and pathological conditions.
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Regeneración Ósea , Hidrogeles , Metilcelulosa , Hidrogeles/química , Hidrogeles/farmacología , Regeneración Ósea/efectos de los fármacos , Metilcelulosa/química , Animales , Inyecciones , Fosfatos de Calcio/química , Fosfatos de Calcio/farmacología , Grafito/química , Bovinos , Proliferación Celular/efectos de los fármacos , Osteogénesis/efectos de los fármacos , HumanosRESUMEN
This paper reports the synthesis, characterization and inâ vivo application of water-soluble supramolecular contrast agents (Mw: 5-5.6â kDa) for MRI obtained from ß-cyclodextrin functionalized with different kinds of nitroxide radicals, both with piperidine structure (CD2 and CD3) and with pyrrolidine structure (CD4 and CD5). As to the stability of the radicals in presence of ascorbic acid, CD4 and CD5 have low second order kinetic constants (≤0.05â M-1 s-1 ) compared to CD2 (3.5â M-1 s-1 ) and CD3 (0.73â M-1 s-1 ). Relaxivity (r1 ) measurements on compounds CD3-CD5 were carried out at different magnetic field strength (0.7, 3, 7 and 9.4 T). At 0.7â T, r1 values comprised between 1.5â mM-1 s-1 and 1.9â mM-1 s-1 were found while a significant reduction was observed at higher fields (r1 ≈0.6-0.9â mM-1 s-1 at 9.4 T). Tests inâ vitro on HEK293 human embryonic kidney cells, L929 mouse fibroblasts and U87 glioblastoma cells indicated that all compounds were non-cytotoxic at concentrations below 1â µmol mL-1 . MRI inâ vivo was carried out at 9.4â T on glioma-bearing rats using the compounds CD3-CD5. The experiments showed a good lowering of T1 relaxation in tumor with a retention of the contrast for at least 60 mins confirming improved stability also inâ vivo conditions.
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Medios de Contraste , Ciclodextrinas , Ratones , Ratas , Humanos , Animales , Medios de Contraste/toxicidad , Medios de Contraste/química , Células HEK293 , Imagen por Resonancia Magnética/métodos , Oxidación-ReducciónRESUMEN
According to statistics and future predictions, meat consumption will increase in the coming years. Considering both the environmental impact of intensive livestock farming and the importance of protecting animal welfare, the necessity of finding alternative strategies to satisfy the growing meat demand is compelling. Biotechnologies are responding to this demand by developing new strategies for producing meat in vitro. The manufacturing of cultured meat has faced criticism concerning, above all, the practical issues of culturing together different cell types typical of meat that are partly responsible for meat's organoleptic characteristics. Indeed, the existence of a cross talk between adipose and muscle cells has critical effects on the outcome of the co-culture, leading to a general inhibition of myogenesis in favor of adipogenic differentiation. This review aims to clarify the main mechanisms and the key molecules involved in this cross talk and provide an overview of the most recent and successful meat culture 3D strategies for overcoming this challenge, focusing on the approaches based on farm-animal-derived cells.
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Ganado , Carne , Animales , Técnicas de Cocultivo , Fenómenos Fisiológicos Celulares , Células MuscularesRESUMEN
Mucoadhesive buccal patches are dosage forms promising for successful drug delivery. They show the distinctive advantages of long residence time on the oral mucosa and increased in situ drug bioavailability. In this context, electrophoretic deposition (EPD) of chitosan (CS) has been demonstrated as a simple and easily tunable technique to produce mucoadhesive buccal patches. However, CS-based buccal patches may suffer from weak mucoadhesion, which can impair their therapeutic effect. In this work, methylcellulose (MC), a widely investigated biopolymer in the biomedical area, was exploited to increase the mucoadhesive characteristic of pristine CS patches. CS-MC patches were obtained in a one-pot process via EPD, and the possibility of incorporating gentamicin sulfate (GS) as a model of a broad-spectrum antibiotic in the so-obtained patches was investigated. The resulting CS-MC patches showed high stability in a water environment and superior mucoadhesive characteristic (σadh = 0.85 ± 0.26 kPa, Wadh = 1192.28 ± 602.36 Pa mm) when compared with the CS control samples (σadh = 0.42 ± 0.22 kPa, Wadh = 343.13 ± 268.89 Pa mm), due to both the control of the patch porosity and the bioadhesive nature of MC. Furthermore, GS-loaded patches showed no in vitro cytotoxic effects by challenging L929 cells with material extracts and noteworthy antibacterial activity on both Gram-positive and Gram-negative bacterial strains.
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Quitosano , Metilcelulosa , Sistemas de Liberación de Medicamentos/métodos , Disponibilidad Biológica , BocaRESUMEN
The development of 3D scaffold-based models would represent a great step forward in cancer research, offering the possibility of predicting the potential in vivo response to targeted anticancer or anti-angiogenic therapies. As regards, 3D in vitro models require proper materials, which faithfully recapitulated extracellular matrix (ECM) properties, adequate cell lines, and an efficient vascular network. The aim of this work is to investigate the possible realization of an in vitro 3D scaffold-based model of adipose tissue, by incorporating decellularized 3D plant structures within the scaffold. In particular, in order to obtain an adipose matrix capable of mimicking the composition of the adipose tissue, methacrylated gelatin (GelMA), UV photo-crosslinkable, was selected. Decellularized fennel, wild fennel and, dill leaves have been incorporated into the GelMA hydrogel before crosslinking, to mimic a 3D channel network. All leaves showed a loss of pigmentation after the decellularization with channel dimensions ranging from 100 to 500 µm up to 3 µm, comparable with those of human microcirculation (5-10 µm). The photo-crosslinking process was not affected by the embedded plant structures in GelMA hydrogels. In fact, the weight variation test, performed on hydrogels with or without decellularized leaves showed a weight loss in the first 96 h, followed by a stability plateau up to 5 weeks. No cytotoxic effects were detected comparing the three prepared GelMA/D-leaf structures; moreover, the ability of the samples to stimulate differentiation of 3T3-L1 preadipocytes in mature adipocytes was investigated, and cells were able to grow and proliferate in the structure, colonizing the entire microenvironment and starting to differentiate. The developed GelMA hydrogels mimicked adipose tissue together with the incorporated plant structures seem to be an adequate solution to ensure an efficient vascular system for a 3D in vitro model. The obtained results showed the potentiality of the innovative proposed approach to mimic the tumoral microenvironment in 3D scaffold-based models.
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Cellulose represents a low cost, abundant, and renewable polysaccharide with great versatility; it has a hierarchical structure composed of nanofibers with high aspect ratio (3-4 nm wide, hundreds of µm long). TEMPO-mediated oxidation represents one of the most diffused methods to obtain cellulose nanofibers (CNFs): It is possible to obtain physically crosslinked hydrogels by means of divalent cation addition. The presence of inorganic components, such as calcium phosphates (CaP), can improve not only their mechanical properties but also the bioactivity of the gels. The aim of this work is to design and characterize a TEMPO-oxidized cellulose nanofibers (TOCNFs) injectable hydrogel embedded with inorganic particles, CaP and CaP-GO, for bone tissue regeneration. Inorganic particles act as physical crosslinkers, as proven by rheological characterization, which reported an increase in mechanical properties. The average load value registered in injection tests was in the range of 1.5-4.4 N, far below 30 N, considered a reasonable injection force upper limit. Samples were stable for up to 28 days and both CaP and CaP-GO accelerate mineralization as suggested by SEM and XRD analysis. No cytotoxic effects were shown on SAOS-2 cells cultured with eluates. This work demonstrated that the physicochemical properties of TOCNFs-based dispersions could be enhanced and modulated through the addition of the inorganic phases, maintaining the injectability and bioactivity of the hydrogels.
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Decellularized tissues are a valid alternative as tissue engineering scaffolds, thanks to the three-dimensional structure that mimics native tissues to be regenerated and the biomimetic microenvironment for cells and tissues growth. Despite decellularized animal tissues have long been used, plant tissue decellularized scaffolds might overcome availability issues, high costs and ethical concerns related to the use of animal sources. The wide range of features covered by different plants offers a unique opportunity for the development of tissue-specific scaffolds, depending on the morphological, physical and mechanical peculiarities of each plant. Herein, three different plant tissues (i.e., apple, carrot, and celery) were decellularized and, according to their peculiar properties (i.e., porosity, mechanical properties), addressed to regeneration of adipose tissue, bone tissue and tendons, respectively. Decellularized apple, carrot and celery maintained their porous structure, with pores ranging from 70 to 420 µm, depending on the plant source, and were stable in PBS at 37°C up to 7 weeks. Different mechanical properties (i.e., Eapple = 4 kPa, Ecarrot = 43 kPa, Ecelery = 590 kPa) were measured and no indirect cytotoxic effects were demonstrated in vitro after plants decellularization. After coating with poly-L-lysine, apples supported 3T3-L1 preadipocytes adhesion, proliferation and adipogenic differentiation; carrots supported MC3T3-E1 pre-osteoblasts adhesion, proliferation and osteogenic differentiation; celery supported L929 cells adhesion, proliferation and guided anisotropic cells orientation. The versatile features of decellularized plant tissues and their potential for the regeneration of different tissues are proved in this work.
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Bioactive glasses (BGs), due to their ability to influence osteogenic cell functions, have become attractive materials to improve loaded and unloaded bone regeneration. BG systems can be easily doped with several metallic ions (e.g., Ag, Sr, Cu, Nb) in order to confer antibacterial properties. In particular, Nb, when compared with other metal ions, has been reported to be less cytotoxic and possess the ability to enhance mineralization process in human osteoblast populations. In this study, we co-deposited, through one-pot electrophoretic deposition (EPD), chitosan (CS), gelatin (GE) and a modified BG containing Nb to obtain substrates with antibacterial activity for unloaded bone regeneration. Self-standing composite scaffolds, with a defined porosity (15-90 µm) and homogeneous dispersion of BGs were obtained. TGA analysis revealed a BG loading of about 10% in the obtained scaffolds. The apatite formation ability of the scaffolds was evaluated in vitro in simulated body fluid (SBF). SEM observations, XRD and FT-IR spectra showed a slow (21-28 days) yet effective nucleation of CaP species on BGs. In particular, FT-IR peak around 603 cm-1 and XRD peak at 2θ = 32°, denoted the formation of a mineral phase after SBF immersion. In vitro biological investigation revealed that the release of Nb from composite scaffolds had no cytotoxic effects. Interestingly, BG-doped Nb scaffolds displayed antibacterial properties, reducing S. lutea and E. coli growth of ≈60% and ≈50%, respectively. Altogether, the obtained results disclose the produced composite scaffolds as promising materials with inherent antibacterial activity for bone tissue engineering applications.
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Regeneración Ósea/fisiología , Cerámica/química , Quitosano/química , Vidrio/química , Niobio/química , Materiales Biocompatibles , Línea Celular Tumoral , Electroforesis , Gelatina , Humanos , Concentración de Iones de Hidrógeno , Ensayo de Materiales , Microscopía Electrónica de Rastreo , Osteosarcoma , Espectroscopía Infrarroja por Transformada de Fourier , Andamios del TejidoRESUMEN
Stable hydrogels with tunable rheological properties were prepared by adding Ca2+ ions to aqueous dispersions of 2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO)-oxidized and ultra-sonicated cellulose nanofibers (TOUS-CNFs). The gelation occurred by interaction among polyvalent cations and the carboxylic units introduced on TOUS-CNFs during the oxidation process. Both dynamic viscosity values and pseudoplastic rheological behaviour increased by increasing the Ca2+ concentration, confirming the cross-linking action of the bivalent cation. The hydrogels were proved to be suitable controlled release systems by measuring the diffusion coefficient of a drug model (ibuprofen, IB) by high-resolution magic angle spinning (HR-MAS) nuclear magnetic resonance (NMR) spectroscopy. IB was used both as free molecule and as a 1:1 pre-formed complex with ß-cyclodextrin (IB/ß-CD), showing in this latter case a lower diffusion coefficient. Finally, the cytocompatibility of the TOUS-CNFs/Ca2+ hydrogels was demonstrated in vitro by indirect and direct tests conducted on a L929 murine fibroblast cell line, achieving a percentage number of viable cells after 7 days higher than 70%.
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Clobetasol propionate (CP) is a high-potency corticosteroid, representing the standard of care for the symptomatic treatment of different skin disorders as well as oral mucosal diseases. Several topical delivery systems are available for treating oral lesions, but the ideal one is still lacking. In this work, we propose a novel class of chitosan (CS) patches, loaded with CP, for the topical treatment of inflammatory chronic oral diseases. Chitosan patches have been fabricated via electrophoretic deposition (EPD), by using a one-pot approach in order to load controlled quantity of CP. Optimized structures showed a water uptake in the range of 200-360% and mechanical properties that allow the design of flexible patches in wet state (E = 0.6 MPa and σbr = 0.55 MPa). Ultraviolet-visible (UV-Vis) spectroscopy was used for the evaluation of both loading and release profile of CP in CS patches. The CP loading has been tuned by adjusting CP concentration in deposition bath-in the range 0.002-0.12 mg cm-2-while releasing curves show an in vitro CP burst of about 80% in the first two hours. Overall, the obtained properties paved the way for the application of this new class of patches for the local oral release of CP.
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Quitosano/química , Quitosano/farmacocinética , Clobetasol/administración & dosificación , Sistemas de Liberación de Medicamentos , Electroforesis , Parche Transdérmico , Administración Tópica , Clobetasol/farmacocinética , Preparaciones de Acción Retardada , Portadores de Fármacos , Composición de Medicamentos/métodos , Liberación de Fármacos , Electroforesis/métodos , Diseño de Equipo/métodos , Etanol/farmacocinética , Humanos , Piel/efectos de los fármacos , Piel/metabolismo , Enfermedades de la Piel/tratamiento farmacológico , Agua/metabolismo , HumectabilidadRESUMEN
One of the main challenges in the design of scaffolds for cortical bone regeneration is mimicking the highly oriented, hierarchical structure of the native tissue in an efficient, simple, and consistent way. As a possible solution to this challenge, positive replica based on electrophoretic deposition (EPD) was here evaluated as a technique to produce organic/inorganic scaffolds with oriented micro-porosities mimicking Haversian canals diameter and spacing. Two different sizes of 45S5 bioactive glass (BG) powders were chosen as inclusions and loaded in a chitosan matrix via EPD on micro-patterned cathodes. Self-standing chitosan scaffolds, with a homogeneous dispersion of BG particles and regularly-oriented micro-channels (Ï = 380 ± 50 µm, inter-channel spacing = 600 ± 40 µm), were obtained. In vitro analysis in simulated body fluid (SBF) revealed the ability to induce a deposition of a homogenous layer of hydroxyapatite (HA), with Ca/P nucleation reactions appearing kinetically favored by smaller BG particles. Cell interaction with hybrid scaffolds was evaluated in vitro with bone osteosarcoma cells (SAOS-2). The osteoconductive potential of EPD structures was assessed by evaluating cells proliferation, viability and scaffold colonization. Results indicate that EPD is a simple yet extremely effective technique to prepare composite micro-patterned structures and can represent a platform for the development of a new generation of bone scaffolds. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2019.
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Cerámica/química , Quitosano/química , Vidrio/química , Andamios del Tejido/química , Líquidos Corporales/química , Línea Celular Tumoral , Supervivencia Celular , ADN/metabolismo , Electroforesis , Humanos , Termogravimetría , Difracción de Rayos XRESUMEN
Advances in regenerative medicine and in modern biomedical therapies are fast evolving and set goals causing an upheaval in the field of materials science. This review discusses recent developments involving the use of biopolymers as smart materials, in terms of material properties and stimulus-responsive behavior, in the presence of environmental physico-chemical changes. An overview on the transformations that can be triggered in natural-based polymeric systems (sol-gel transition, polymer relaxation, cross-linking, and swelling) is presented, with specific focus on the benefits these materials can provide in biomedical applications.
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Órganos Artificiales , Biopolímeros , Diseño de Equipo , Humanos , Medicina RegenerativaRESUMEN
Neo-vascularization is a key factor in tissue regeneration within porous scaffolds. Here, we tested the hypothesis that micro-patterned scaffolds, with precisely-designed, open micro-channels, might help endothelial cells to produce intra-scaffold vascular networks. Three series of micro-patterned scaffolds were produced via electrochemical replica-deposition of chitosan and cross-linking. All had regularly-oriented micro-channels (Ï 500 µm), which differed for the inter-channel spacing, at 600, 700, or 900 µm, respectively. Random-pore scaffolds, using the same technique, were taken as controls. Physical-mechanical characterization revealed high water uptake and favorable elastic mechanical behavior for all scaffolds, slightly reduced in the presence of cross-linking and enhanced with the 700 µm-spaced micro-pattern. At MTT assay, mouse endothelial cell viability was >90% at day 1, 3 and 7, confirmed by visual examination with scanning electron microscopy (SEM). Intra-scaffold cell density, at fluorescence analysis, was higher for the 600 µm-spaced and the 700 µm-spaced micro-patterns over the others. The 700 µm-spaced scaffold was selected for the in vivo testing, to be compared to the random-pore one. Neither type produced an inflammatory reaction; both showed excellent tissue ingrowth. Micro-patterned scaffolds enhanced neo-vascularization, demonstrated by immunofluorescent, semi-quantitative analyses. These findings support the use of micro-patterned porous scaffolds, with adequately spaced micro-channels, to promote neo-vascularization.
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Neovascularización Fisiológica , Andamios del Tejido/química , Animales , Materiales Biocompatibles/efectos adversos , Materiales Biocompatibles/química , Recuento de Células , Proliferación Celular , Supervivencia Celular , Quitosano/efectos adversos , Quitosano/química , Quitosano/inmunología , Reactivos de Enlaces Cruzados , Técnicas Electroquímicas/métodos , Células Endoteliales/citología , Células Endoteliales/fisiología , Femenino , Masculino , Ensayo de Materiales , Ratones , Ratones Endogámicos C57BL , Microscopía Electrónica de Rastreo , Microtecnología/métodos , Porosidad , Regeneración/fisiología , Andamios del Tejido/efectos adversosRESUMEN
Methylcellulose (MC), a water-soluble polymer derived from cellulose, was investigated as a possible temporary substrate having thermo-responsive properties favorable for cell culturing. MC-based hydrogels were prepared by a dispersion technique, mixing MC powder (2, 4, 6, 8, 10, 12 % w/v) with selected salts (sodium sulphate, Na2SO4), sodium phosphate, calcium chloride, or phosphate buffered saline, to evaluate the influence of different compositions on the thermo-responsive behavior. The inversion test was used to determine the gelation temperatures of the different hydrogel compositions; thermo-mechanical properties and thermo-reversibility of the MC hydrogels were investigated by rheological analysis. Gelation temperatures and rheological behavior depended on the MC concentration and type and concentration of salt used in hydrogel preparation. In vitro cytotoxicity tests, performed using L929 mouse fibroblasts, showed no toxic release from all the tested hydrogels. Among the investigated compositions, the hydrogel composed of 8 % w/v MC with 0.05 M Na2SO4 had a thermo-reversibility temperature at 37 °C. For that reason, this formulation was thus considered to verify the possibility of inducing in vitro spontaneous detachment of cells previously seeded on the hydrogel surface. A continuous cell layer (cell sheet) was allowed to grow and then detached from the hydrogel surface without the use of enzymes, thanks to the thermo-responsive behavior of the MC hydrogel. Immunofluorescence observation confirmed that the detached cell sheet was composed of closely interacting cells.
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Proliferación Celular/fisiología , Fibroblastos/fisiología , Hidrogeles/química , Metilcelulosa/química , Animales , Materiales Biocompatibles , Técnicas de Cultivo de Célula , Ratones , Células 3T3 NIH , Reología , TemperaturaRESUMEN
Large bone defects are challenging to heal, and often require an osteoconductive and stable support to help the repair of damaged tissue. Bioglass-based scaffolds are particularly promising for this purpose due to their ability to stimulate bone regeneration. However, processing technologies adopted so far do not allow for the synthesis of scaffolds with suitable mechanical properties. Also, conventional sintering processes result in glass de-vitrification, which generates concerns about bioactivity. In this work, we studied the bioactivity and the mechanical properties of Bioglass(®) based scaffolds, produced via a powder technology inspired process. The scaffolds showed compressive strengths in the range of 5-40 MPa, i.e. in the upper range of values reported so far for these materials, had tunable porosity, in the range between 55 and 77%, and pore sizes that are optimal for bone tissue regeneration (100-500 µm). We immersed the scaffolds in simulated body fluid (SBF) for 28 d and analyzed the evolution of the scaffold mechanical properties and microstructure. Even if, after sintering, partial de-vitrification occurred, immersion in SBF caused ion release and the formation of a Ca-P coating within 2 d, which reached a thickness of 10-15 µm after 28 d. This coating contained both hydroxyapatite and an amorphous background, indicating microstructural amorphization of the base material. Scaffolds retained a good compressive strength and structural integrity also after 28 d of immersion (6 MPa compressive strength). The decrease in mechanical properties was mainly related to the increase in porosity, caused by its dissolution, rather than to the amorphization process and the formation of a Ca-P coating. These results suggest that Bioglass(®) based scaffolds produced via powder metallurgy-inspired technique are excellent candidates for bone regeneration applications.
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Sustitutos de Huesos/síntesis química , Cerámica/química , Vidrio/química , Andamios del Tejido , Fuerza Compresiva , Módulo de Elasticidad , Diseño de Equipo , Análisis de Falla de Equipo , Dureza , Calefacción/métodos , Ensayo de Materiales , Metalurgia/métodos , Polvos , Estrés Mecánico , Resistencia a la TracciónRESUMEN
Cathodic electrodeposition is a bottom up process that is emerging as a simple yet efficient strategy to engineer thin polymeric films with well-defined physicochemical properties. In particular, this technique offers the distinctive advantage of an easy control over composition, thickness, and morphology of the films by simply adjusting treatment parameters. In this work, cathodic electrodeposition was exploited to engender blends composed by chitosan (CH) and poly-ethylene-oxide (PEO) with different weight ratios. The physicochemical and nanomechanical properties of the resulting films were successively characterized by integrating Raman and Fourier-transform infrared (FT-IR) spectroscopy with Atomic Force Microscopy (AFM). Our findings demonstrate that electro-deposition is an effective technique for the co-deposition of CH:PEO blends. Moreover, spectroscopic and AFM analyses correlated the physicochemical (i.e. structural organization, bond formation and cross-linking) and nanomechanical properties of the blends to the PEO content, ultimately unveiling the molecular interactions and mechanisms involved in the cathodic deposition of CH:PEO films.
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INTRODUCTION AND HYPOTHESIS: To test in vitro and in vivo the capability of mesh materials to act as scaffolds for rat-derived mesenchymal stem cells (rMSCs) and to compare inflammatory response and collagen characteristics of implant materials, either seeded or not with rMSCs. METHODS: rMSCs isolated from rat bone marrow were seeded and cultured in vitro on four different implant materials. Implants showing the best rMSC proliferation rate were selected for the in vivo experiment. Forty-eight adult female Sprague-Dawley rats were randomly divided into two treatment groups. The implant of interest-either seeded or not with rMSCs-was laid and fixed over the muscular abdominal wall. Main outcome measures were: in vitro, proliferation of rMSCs on selected materials; in vivo, the occurrence of topical complications, the evaluation of systemic and local inflammatory response and examination of the biomechanical properties of explants. RESULTS: Surgisis and Pelvitex displayed the best cell growth in vitro. At 90 days in the rat model, rMSCs were related to a lower count of neutrophil cells for Pelvitex and a greater organisation and collagen amount for Surgisis. At 7 days Surgisis samples seeded with rMSCs displayed higher breaking force and stiffness. CONCLUSIONS: The presence of rMSCs reduced the systemic inflammatory response on synthetic implants and improved collagen characteristics at the interface between biological grafts and native tissues. rMSCs enhanced the stripping force on biological explants.
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Células Madre Mesenquimatosas/fisiología , Mallas Quirúrgicas , Andamios del Tejido , Dermis Acelular/efectos adversos , Animales , Materiales Biocompatibles/efectos adversos , Proliferación Celular , Células Cultivadas , Colágeno/efectos adversos , Colágeno/metabolismo , Colágeno/ultraestructura , Elasticidad , Femenino , Inflamación/etiología , Recuento de Leucocitos , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/citología , Neutrófilos , Polipropilenos/efectos adversos , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Mallas Quirúrgicas/efectos adversos , Resistencia a la Tracción , Andamios del Tejido/efectos adversosRESUMEN
Chitosan (CS) is a versatile biopolymer whose morphological and chemico-physical properties can be designed for a variety of biomedical applications. Taking advantage of its electrolytic nature, cathodic polarization allows CS deposition on electrically conductive substrates, resulting in thin porous structures with tunable morphology. Here we propose an easy method to obtain CS membranes with highly oriented micro-channels for tissue engineering applications, relying on simple control of process parameters and cathodic substrate geometry. Cathodic deposition was performed on two different aluminum grids in galvanostatic conditions at 6.25 mA cm(-2) from CS solution [1g L(-1)] in acetic acid (pH 3.5). Self-standing thin scaffolds were cross linked either with genipin or epichlorohydrin, weighted, and observed by optical and electron microscopy. Swelling properties at pH 5 and pH 7.4 have been also investigated and tensile tests performed on swollen samples at room temperature. Finally, direct and indirect assays have been performed to evaluate the cytotoxicity at 24 and 72 h. Thin scaffolds with two different oriented porosities (1000 µm and 500 µm) have been successfully fabricated by electrochemical techniques. Both cross-linking agents did not affected the mechanical properties and cytocompatibility of the resulting structures. Depending on the pH, these structures show interesting swelling properties that can be exploited for drug delivery systems. Moreover, thanks to the possibility of controlling the porosity and the micro-channel orientation, they should be used for the regeneration of tissues requiring a preferential cells orientation, e.g., cardiac patches or ligament regeneration.
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Materiales Biocompatibles/química , Quitosano/química , Técnicas Electroquímicas/métodos , Andamios del Tejido/química , Animales , Fenómenos Químicos , Células Endoteliales/efectos de los fármacos , Epiclorhidrina/química , Concentración de Iones de Hidrógeno , Iridoides/química , Ratones , Microscopía Electrónica de Rastreo , Porosidad , Regeneración , Ingeniería de Tejidos/métodosRESUMEN
In cementless fixation of metallic prostheses, bony ingrowth onto the implant surface is often promoted by osteoconductive plasma-sprayed hydroxyapatite coatings. The present work explores the use of the innovative High Velocity Suspension Flame Spraying (HVSFS) process to coat Ti substrates with thin homogeneous hydroxyapatite coatings. The HVSFS hydroxyapatite coatings studied were dense, 27-37µm thick, with some transverse microcracks. Lamellae were sintered together and nearly unidentifiable, unlike conventional plasma-sprayed hydroxyapatite. Crystallinities of 10%-70% were obtained, depending on the deposition parameters and the use of a TiO2 bond coat. The average hardness of layers with low (<24%) and high (70%) crystallinity was ≈3.5GPa and ≈4.5GPa respectively. The distributions of hardness values, all characterised by Weibull modulus in the 5-7 range, were narrower than that of conventional plasma-sprayed hydroxyapatite, with a Weibull modulus of ≈3.3. During soaking in simulated body fluid, glassy coatings were progressively resorbed and replaced by a new, precipitated hydroxyapatite layer, whereas coatings with 70% crystallinity were stable up to 14days of immersion. The interpretation of the precipitation behaviour was also assisted by surface charge assessments, performed through Z-potential measurements. During in vitro tests, HA coatings showed no cytotoxicity towards the SAOS-2 osteoblast cell line, and surface cell proliferation was comparable with proliferation on reference polystyrene culture plates.
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Durapatita/química , Ensayo de Materiales/métodos , Temperatura , Líquidos Corporales/química , Comunicación Celular/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Materiales Biocompatibles Revestidos/química , Materiales Biocompatibles Revestidos/farmacología , Cristalización , Análisis Diferencial Térmico , Durapatita/farmacología , Módulo de Elasticidad/efectos de los fármacos , Dureza/efectos de los fármacos , Humanos , Microscopía Electrónica de Rastreo , Gases em Plasma/química , Polvos , Espectrometría Raman , Electricidad Estática , Suspensiones , Resistencia a la Tracción/efectos de los fármacos , Termogravimetría , Difracción de Rayos XRESUMEN
Industrial manufacturing of prosthesis components could take significant advantage by the introduction of new, cost-effective manufacturing technologies with near net-shape capabilities, which have been developed during the last years to fulfill the needs of different technological sectors. Among them, metal injection molding (MIM) appears particularly promising for the production of orthopedic arthroplasty components with significant cost saving. These new manufacturing technologies, which have been developed, however, strongly affect the chemicophysical structure of processed materials and their resulting properties. In order to investigate this relationship, here we evaluated the effects on electrochemical properties, ion release, and in vitro response of medical grade CoCrMo alloy processed via MIM compared to conventional processes. MIM of the CoCrMo alloy resulted in coarser polygonal grains, with largely varying sizes; however, these microstructural differences between MIM and forged/cast CoCrMo alloys showed a negligible effect on electrochemical properties. Passive current densities values observed were 0.49 µA cm(-2) for MIM specimens and 0.51 µA cm(-2) for forged CoCrMo specimens, with slightly lower transpassive potential in the MIM case; open circuit potential and Rp stationary values showed no significant differences. Moreover, in vitro biocompatibility tests resulted in cell viability levels not significantly different for MIM and conventionally processed alloys. Although preliminary, these results support the potential of MIM technology for the production of CoCrMo components of implantable devices.