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BACKGROUND: Assisted Reproductive Technologies (ARTs) have been validated in human and animal to solve reproductive problems such as infertility, aging, genetic selection/amplification and diseases. The persistent gap in ART biomedical applications lies in recapitulating the early stage of ovarian folliculogenesis, thus providing protocols to drive the large reserve of immature follicles towards the gonadotropin-dependent phase. Tissue engineering is becoming a concrete solution to potentially recapitulate ovarian structure, mostly relying on the use of autologous early follicles on natural or synthetic scaffolds. Based on these premises, the present study has been designed to validate the use of the ovarian bioinspired patterned electrospun fibrous scaffolds fabricated with poly(ε-caprolactone) (PCL) for multiple preantral (PA) follicle development. METHODS: PA follicles isolated from lamb ovaries were cultured on PCL scaffold adopting a validated single-follicle protocol (Ctrl) or simulating a multiple-follicle condition by reproducing an artificial ovary engrafted with 5 or 10 PA (AO5PA and AO10PA). The incubations were protracted for 14 and 18 days before assessing scaffold-based microenvironment suitability to assist in vitro folliculogenesis (ivF) and oogenesis at morphological and functional level. RESULTS: The ivF outcomes demonstrated that PCL-scaffolds generate an appropriate biomimetic ovarian microenvironment supporting the transition of multiple PA follicles towards early antral (EA) stage by supporting follicle growth and steroidogenic activation. PCL-multiple bioengineering ivF (AO10PA) performed in long term generated, in addition, the greatest percentage of highly specialized gametes by enhancing meiotic competence, large chromatin remodeling and parthenogenetic developmental competence. CONCLUSIONS: The study showcased the proof of concept for a next-generation ART use of PCL-patterned scaffold aimed to generate transplantable artificial ovary engrafted with autologous early-stage follicles or to advance ivF technologies holding a 3D bioinspired matrix promoting a physiological long-term multiple PA follicle protocol.
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Folículo Ovárico , Poliésteres , Ingeniería de Tejidos , Andamios del Tejido , Femenino , Folículo Ovárico/crecimiento & desarrollo , Folículo Ovárico/citología , Andamios del Tejido/química , Animales , Poliésteres/química , Ingeniería de Tejidos/métodos , Ovinos , Ovario/crecimiento & desarrollo , Ovario/citología , Oogénesis/fisiología , Oogénesis/efectos de los fármacos , Bioingeniería/métodos , Técnicas Reproductivas Asistidas , Fertilización In Vitro/métodosRESUMEN
In vitro maturation (IVM) is not a routine assisted reproductive technology (ART) for oocytes collected from early antral (EA) follicles, a large source of potentially available gametes. Despite substantial improvements in IVM in the past decade, the outcomes remain low for EA-derived oocytes due to their reduced developmental competences. To optimize IVM for ovine EA-derived oocytes, a three-dimensional (3D) scaffold-mediated follicle-enclosed oocytes (FEO) system was compared with a validated cumulus-oocyte complex (COC) protocol. Gonadotropin stimulation (eCG and/or hCG) and/or somatic cell coculture (ovarian vs. extraovarian-cell source) were supplied to both systems. The maturation rate and parthenogenetic activation were significantly improved by combining hCG stimulation with ovarian surface epithelium (OSE) cells coculture exclusively on the FEO system. Based on the data, the paracrine factors released specifically from OSE enhanced the hCG-triggering of oocyte maturation mechanisms by acting through the mural compartment (positive effect on FEO and not on COC) by stimulating the EGFR signaling. Overall, the FEO system performed on a developed reproductive scaffold proved feasible and reliable in promoting a synergic cytoplasmatic and nuclear maturation, offering a novel cultural strategy to widen the availability of mature gametes for ART.
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Técnicas de Maduración In Vitro de los Oocitos , Ingeniería de Tejidos , Femenino , Ovinos , Animales , Humanos , Técnicas de Cocultivo , Técnicas de Maduración In Vitro de los Oocitos/métodos , Oocitos/metabolismo , Gonadotropina Coriónica/farmacología , Gonadotropina Coriónica/metabolismo , EpitelioRESUMEN
Mesoporous silica-based nanoparticles (MSNs) are considered promising drug carriers because of their ordered pore structure, which permits high drug loading and release capacity. The dissolution of Si and Ca from MSNs can trigger osteogenic differentiation of stem cells towards extracellular matrix calcification, while Mg and Sr constitute key elements of bone biology and metabolism. The aim of this study was the synthesis and characterization of sol-gel-derived MSNs co-doped with Ca, Mg and Sr. Their physico-chemical properties were investigated by X-ray diffraction (XRD), scanning electron microscopy with energy dispersive X-ray analysis (SEM/EDX), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), X-ray fluorescence spectroscopy (XRF), Brunauer Emmett Teller and Brunauer Joyner Halenda (BET/BJH), dynamic light scattering (DLS) and ζ-potential measurements. Moxifloxacin loading and release profiles were assessed with high performance liquid chromatography (HPLC) cell viability on human periodontal ligament fibroblasts and their hemolytic activity in contact with human red blood cells (RBCs) at various concentrations were also investigated. Doped MSNs generally retained their textural characteristics, while different compositions affected particle size, hemolytic activity and moxifloxacin loading/release profiles. All co-doped MSNs revealed the formation of hydroxycarbonate apatite on their surface after immersion in simulated body fluid (SBF) and promoted mitochondrial activity and cell proliferation.
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Sistemas de Liberación de Medicamentos , Moxifloxacino/farmacología , Nanopartículas/química , Ingeniería de Tejidos , Proliferación Celular/efectos de los fármacos , Dispersión Dinámica de Luz , Humanos , Magnesio/química , Microscopía Electrónica de Rastreo , Moxifloxacino/química , Osteogénesis/efectos de los fármacos , Porosidad , Dióxido de Silicio/química , Difracción de Rayos XRESUMEN
In a century when environmental pollution is a major issue, polymers issued from bio-based monomers have gained important interest, as they are expected to be environment-friendly, and biocompatible, with non-toxic degradation products. In parallel, hyperbranched polymers have emerged as an easily accessible alternative to dendrimers with numerous potential applications. Glycerol (Gly) is a natural, low-cost, trifunctional monomer, with a production expected to grow significantly, and thus an excellent candidate for the synthesis of hyperbranched polyesters for pharmaceutical and biomedical applications. In the present article, we review the synthesis, properties, and applications of glycerol polyesters of aliphatic dicarboxylic acids (from succinic to sebacic acids) as well as the copolymers of glycerol or hyperbranched polyglycerol with poly(lactic acid) and poly(ε-caprolactone). Emphasis was given to summarize the synthetic procedures (monomer molar ratio, used catalysts, temperatures, etc.,) and their effect on the molecular weight, solubility, and thermal and mechanical properties of the prepared hyperbranched polymers. Their applications in pharmaceutical technology as drug carries and in biomedical applications focusing on regenerative medicine are highlighted.
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Materiales Biocompatibles , Dendrímeros , Portadores de Fármacos , Glicerol , Poliésteres , Polímeros , Materiales Biocompatibles/síntesis química , Materiales Biocompatibles/química , Materiales Biocompatibles/uso terapéutico , Dendrímeros/síntesis química , Dendrímeros/química , Dendrímeros/uso terapéutico , Portadores de Fármacos/síntesis química , Portadores de Fármacos/química , Portadores de Fármacos/uso terapéutico , Glicerol/síntesis química , Glicerol/química , Glicerol/uso terapéutico , Humanos , Poliésteres/síntesis química , Poliésteres/química , Poliésteres/uso terapéutico , Polímeros/síntesis química , Polímeros/química , Polímeros/uso terapéutico , Medicina RegenerativaRESUMEN
Mesoporous silica nanoparticles were synthesized using a microemulsion-assisted sol-gel method, and calcium, gallium or a combination of both, were used as dopants. The influence of these metallic ions on the physicochemical properties of the nanoparticles was investigated by scanning and transmission electron microscopy, as well as N2 adsorption-desorption methods. The presence of calcium had a significant impact on the morphology and textural features of the nanoparticles. The addition of calcium increased the average diameter of the nanoparticles from 80 nm to 150 nm, while decreasing their specific surface area from 972 m2/g to 344 m2/g. The nanoparticles of all compositions were spheroidal, with a disordered mesoporous structure. An ion release study in cell culture medium demonstrated that gallium was released from the nanoparticles in a sustained manner. In direct contact with concentrations of up to 100 µg/mL of the nanoparticles, gallium-containing nanoparticles did not exhibit cytotoxicity towards pre-osteoblast MC3T3-E1 cells. Moreover, in vitro cell culture tests revealed that the addition of gallium to the nanoparticles enhanced osteogenic activity. Simultaneously, the nanoparticles disrupted the osteoclast differentiation of RAW 264.7 macrophage cells. These findings suggest that gallium-containing nanoparticles possess favorable physicochemical properties and biological characteristics, making them promising candidates for applications in bone tissue regeneration, particularly for unphysiological or pathological conditions such as osteoporosis.
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Galio , Nanopartículas , Osteoclastos , Osteogénesis , Galio/química , Galio/farmacología , Animales , Ratones , Osteoclastos/efectos de los fármacos , Nanopartículas/química , Osteogénesis/efectos de los fármacos , Dióxido de Silicio/química , Dióxido de Silicio/farmacología , Células RAW 264.7 , Porosidad , Diferenciación Celular/efectos de los fármacosRESUMEN
In this work, composite electrospun fibers containing innovative bioactive glass nanoparticles were produced and characterized. Poly(ε-caprolactone), benign solvents, and sol-gel B- and Cu-doped bioactive glass powders were used to fabricate fibrous scaffolds. The retention of bioactive glass nanoparticles in the polymer matrix, the electrospinnability of this novel solution and the obtained electrospun composites were extensively characterized. As a result, composite electrospun fibers characterized by biocompatibility, bioactivity, and exhibiting overall properties adequate for both hard and soft tissue engineering applications, have been produced. The addition of these bioactive glass nanoparticles was, indeed, able to impart bioactive properties to the fibers. Cell culture studies show promising results, demonstrating proliferation and growth of cells on the composite fibers. Wettability, degradation rate, and mechanical performance were also tested and are in line with previous results.
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Materiales Biocompatibles , Ingeniería de Tejidos , Ingeniería de Tejidos/métodos , Poliésteres , Polímeros , Vidrio , Andamios del TejidoRESUMEN
Human Adipose-Derived Mesenchymal Stem/Stromal Cells (hAD-MSCs) have great potential for tissue regeneration. Since transplanted hAD-MSCs are likely to be placed in a hypoxic environment, culturing the cells under hypoxic conditions might improve their post-transplantation survival and regenerative performance. The combination of hAD-MSCs and PCL-nHA nanofibers synergically improves the contribution of both components for osteoblast differentiation. In this work, we hypothesized that this biomaterial constitutes a hypoxic environment for hAD-MSCs. We studied the cellular re-arrangement and the subcellular ultrastructure by Transmission Electron Microscopy (TEM) of hAD-MSCs grown into PCL-nHA nanofibers, and we compared them with the same cells grown in two-dimensional cultures, over tissue culture-treated plastic, or glass coverslips. Among the most evident changes, PCL-nHA grown cells showed enlarged mitochondria, and accumulation of glycogen granules, consistent with a hypoxic environment. We observed a 3.5 upregulation (p = 0.0379) of Hypoxia Inducible Factor (HIF)-1A gene expression in PCL-nHA grown cells. This work evidences for the first time intra-cellular changes in three-dimensional compared to two-dimensional cultures, which are adaptive responses of the cells to an environment more closely resembling that of the in vivo niche after transplantation, thus PCL-nHA nanofibers are adequate for hAD-MSCs pre-conditioning.
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Células Madre Mesenquimatosas , Nanofibras , Humanos , Andamios del Tejido/química , Durapatita/química , Durapatita/metabolismo , Poliésteres/química , Materiales Biocompatibles/química , Diferenciación Celular , Nanofibras/química , Ingeniería de Tejidos/métodosRESUMEN
We report the successful preparation and characterization of chitosan-Zn complex (ChiZn) in the form of films, intended to enhance the biological performance of chitosan by the presence of Zn as antibacterial agent and biologically active ion. The influence of Zn chelation on morphology and structure of chitosan was assessed by scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy and infrared spectroscopy. The biodegradability study of ChiZn showed a sustained release of Zn up to 2 mg/mL. No toxic response was observed toward stromal cell line ST-2 in indirect contact with the ChiZn films. The dissolution product of ChiZn showed improved wound closure (88% closure) compared to the positive control group (70% closure). Moreover, ChiZn exhibited antibacterial activity against S. aureus together with a slight increase (~30%) in the secretion of VEGF and moderate decrease in nitric oxide evolution. Our findings indicate that ChiZn could be used as a safe and effective wound healing agent.
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Quitosano , Antibacterianos/química , Antiinflamatorios , Movimiento Celular , Quitosano/química , Quitosano/farmacología , Staphylococcus aureus , Cicatrización de Heridas , Zinc/farmacologíaRESUMEN
Electrospun fibers of shape memory triethoxysilane-terminated poly(epsilon-caprolactone) (PCL-TES) loaded with bioactive glasses (BG) are here presented. Unloaded PCL-TES, as well as PCL/BG nanocomposite fibers, are also considered for comparison. It is proposed that hydrolysis and condensation reactions take place between triethoxysilane groups of the polymer and the silanol groups at the BG particle surface, thus generating additional crosslinking points with respect to those present in the PCL-TES system. The as-spun PCL-TES/BG fibers display excellent shape memory properties, in terms of shape fixity and shape recovery ratios, without the need of a thermal crosslinking treatment. BG particles confer in vitro bioactivity to PCL-based nanocomposite fibers and favor the precipitation of hydroxycarbonate apatite on the fiber surface. Preliminary cytocompatibility tests demonstrate that the addition of BG particles to PCL-based polymer does not inhibit ST-2 cell viability. This novel approach of using bioactive glasses not only for their biological properties, but also for the enhancement of shape memory properties of PCL-based polymers, widens the versatility and suitability of the obtained composite fibers for a huge portfolio of biomedical applications.
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Current assisted reproduction technologies (ART) are insufficient to cover the slice of the population needing to restore fertility, as well as to amplify the reproductive performance of domestic animals or endangered species. The design of dedicated reproductive scaffolds has opened the possibility to better recapitulate the reproductive 3D ovarian environment, thus potentially innovating in vitro folliculogenesis (ivF) techniques. To this aim, the present research has been designed to compare ovine preantral follicles in vitro culture on poly(epsilon-caprolactone) (PCL)-based electrospun scaffolds designed with different topology (Random vs. Patterned fibers) with a previously validated system. The ivF performances were assessed after 14 days under 3D-oil, Two-Step (7 days in 3D-oil and on scaffold), or One-Step PCL protocols (14 days on PCL-scaffold) by assessing morphological and functional outcomes. The results show that Two- and One-Step PCL ivF protocols, when performed on patterned scaffolds, were both able to support follicle growth, antrum formation, and the upregulation of follicle marker genes leading to a greater oocyte meiotic competence than in the 3D-oil system. In conclusion, the One-Step approach could be proposed as a practical and valid strategy to support a synergic follicle-oocyte in vitro development, providing an innovative tool to enhance the availability of matured gametes on an individual basis for ART purposes.
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Caproatos , Andamios del Tejido , Animales , Lactonas , OvinosRESUMEN
Electrospun fibers based on biodegradable polyanionic or polycationic biopolymers are highly beneficial for biomedical applications. In this work, electrospun nanofibers made from poly(epsilon caprolactone) (PCL), chitosan (CS) andκ-carrageenan (κ-C) were successfully fabricated using several mixtures of benign solvents containing formic acid and acetic acid. The addition ofκ-C improved the preparation procedure for the production of PCL/CS fibers by electrospinning. Moreover, a polymer mixture was selected to be stored at -20 °C for one month with the purpose to study the properties of the resulting fiber mat. The results indicated that fiber characteristics were not seriously compromised compared to the ones of those fabricated with the original solution, which represents an important reduction in produced waste. Thus, the interactions that occur between positively and negatively charged hydrophilic polysaccharides might induce higher stability to the linear aliphatic polyester in the polymer mixture. All fiber mats were morphologically, physico-chemically and mechanically characterized, showing average fiber diameters in the nano scale. A direct cell viability assay using ST-2 cells demonstrated cell proliferation after seven days of incubation for all prepared fiber mats, confirming their suitability as potential candidates for bone tissue engineering and wound healing applications.
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Quitosano , Nanofibras , Carragenina , Quitosano/química , Nanofibras/química , Poliésteres/química , Polímeros/química , Ingeniería de Tejidos/métodos , Andamios del Tejido/químicaRESUMEN
BACKGROUND: Aliphatic polyesters are widely used for biomedical, pharmaceutical and environmental applications due to their high biodegradability and cost-effective production. Recently, star and hyperbranched polyesters based on glycerol and ω-carboxy fatty diacids have gained considerable interest. Succinic acid and bio-based diacids similar to glycerol are regarded as safe materials according to the US Food and Drug Administration (FDA). Bioactive glass scaffolds utilized in bone tissue engineering are relatively brittle materials. However, their mechanical properties can be improved by using polymer coatings that can further control their degradation rate, tailor their biocompatibility and enhance their performance. The purpose of this study is to explore a new biopolyester poly(glycerol succinate) (PGSuc) reinforced with mesoporous bioactive nanoparticles (MSNs) as a novel coating material to produce hybrid scaffolds for bone tissue engineering. METHODS: Bioactive glass scaffolds were coated with neat PGSuc, PGSuc loaded with dexamethasone sodium phosphate (DexSP) and PGSuc loaded with DexSP-laden MSNs. The physicochemical, mechanical and biological properties of the scaffolds were also evaluated. RESULTS: Preliminary data are provided showing that polymer coatings with and without MSNs improved the physicochemical properties of the 1393 bioactive glass scaffolds and increased the ALP activity and alizarin red staining, suggesting osteogenic differentiation potential when cultured with adipose-derived mesenchymal stem cells. CONCLUSIONS: PGSuc with incorporated MSNs coated onto 1393 bioactive glass scaffolds could be promising candidates in bone tissue engineering applications.
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Tendon injuries are at the frontier of innovative approaches to public health concerns and sectoral policy objectives. Indeed, these injuries remain difficult to manage due to tendon's poor healing ability ascribable to a hypo-cellularity and low vascularity, leading to the formation of a fibrotic tissue affecting its functionality. Tissue engineering represents a promising solution for the regeneration of damaged tendons with the aim to stimulate tissue regeneration or to produce functional implantable biomaterials. However, any technological advancement must take into consideration the role of the immune system in tissue regeneration and the potential of biomaterial scaffolds to control the immune signaling, creating a pro-regenerative environment. In this context, immunoengineering has emerged as a new discipline, developing innovative strategies for tendon injuries. It aims at designing scaffolds, in combination with engineered bioactive molecules and/or stem cells, able to modulate the interaction between the transplanted biomaterial-scaffold and the host tissue allowing a pro-regenerative immune response, therefore hindering fibrosis occurrence at the injury site and guiding tendon regeneration. Thus, this review is aimed at giving an overview on the role exerted from different tissue engineering actors in leading immunoregeneration by crosstalking with stem and immune cells to generate new paradigms in designing regenerative medicine approaches for tendon injuries.
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Inmunidad , Regeneración/fisiología , Tendones/inmunología , Tendones/fisiología , Ingeniería de Tejidos , Andamios del Tejido/química , Animales , Humanos , InmunomodulaciónRESUMEN
Wound dressings produced by electrospinning exhibit a fibrous structure close to the one of the extracellular matrix of the skin. In this article, electrospinning was used to fabricate fiber mats based on the well-known biopolymers poly(É-caprolactone) (PCL) and methylcellulose (MC) using benign solvents. The blend fiber mats were cross-linked using Manuka honey and additionally used as a biodegradable platform to deliver bioactive glass particles. It was hypothesized that a dual therapeutic effect can be achieved by combining Manuka honey and bioactive glass. Morphological and chemical examinations confirmed the successful production of submicrometric PCL-MC fiber mats containing Manuka honey and bioactive glass particles. The multifunctional fiber mats exhibited improved wettability and suitable mechanical properties (ultimate tensile strength of 3-5 MPa). By performing dissolution tests using simulated body fluid, the improved bioactivity of the fiber mats by the addition of bioactive glass was confirmed. Additionally, cell biology tests using human dermal fibroblasts and human keratinocytes-like HaCaT cells showed the potential of the fabricated composite fiber mats to be used as wound dressing, specially due to the ability to support wound closure influenced by the presence of bioactive glass. Moreover, based on the results of the antibacterial tests, it is apparent that an optimization of the electrospun fiber mats is required to develop suitable wound dressing for the treatment of infected wounds.
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Vendajes , Vidrio/química , Miel , Queratinocitos/metabolismo , Ensayo de Materiales , Metilcelulosa/química , Poliésteres/química , Antibacterianos/química , Línea Celular , HumanosRESUMEN
The aim of this study was to obtain hierarchical scaffolds combining 3D printing and two electrofluidodynamic methods. The multi-layered scaffold is composed by 3D printed struts, electrospun fibers obtained from poly(ϵ-caprolactone) and electrosprayed spheres produced from hydrophobically modified chitosan, namely chitosan grafted with linoleic acid (CHLA). Since CHLA has been used for the first time in the electrospraying (electro dynamic spraying, EDS) process, the formation of spheres needed an optimization process. The EDS process was strongly affected by the solvent mixture composition, concentration of acid used for CHLA dissolution and solution flow rate. By using the optimized electrospraying conditions, uniformly distributed spheres have been obtained, decorating struts and nanofibers. Preliminary biological tests with mouse preosteoblasts (MC3T3-E1) were performed to investigate the effect of the hierarchical scaffold on cell seeding efficacy. Results showed that the hierarchical structure enhances cell seeding efficacy, respect to the 3D printed struts alone, preventing that the cells passed through the struts during the seeding. Moreover, the addition of the electrosprayed nanoparticles does not affect the cell seeding efficiency. The versatility of the proposed structure, with the added value of CHLA nanoparticles decoration could be suitable for several applications in tissue engineering, mainly related to drug delivery systems.
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Quitosano , Nanofibras , Animales , Ratones , Nanofibras/química , Impresión Tridimensional , Ingeniería de Tejidos/métodos , Andamios del Tejido/químicaRESUMEN
In recent years, mesoporous bioactive glass nanoparticles (MBGNPs) have generated great attention in biomedical applications. In this study, cerium and gallium doped MBGNPs were prepared by microemulsion assisted sol-gel method in the binary SiO2-CaO system. MBGNPs with spheroidal and pineal shaped morphology were obtained. Nitrogen sorption analysis elucidated the mesoporous structure of synthesized nanoparticles with high specific surface area. X-ray diffraction analysis confirmed the amorphous nature of the nanoparticles. The chemical compositions of all samples were determined by inductively coupled plasma-optical emission spectrometry (ICP-OES), which revealed that the contents of cerium and gallium could be tailored by adjusting the concentrations of the precursors used for the synthesis. All MBGNPs exhibited in vitro bioactivity when immersed in simulated body fluid, except the particles doped with higher amounts than 1 mol% of cerium. MBGNPs showed antibacterial activity against S. aureus and E. coli without exhibiting cytotoxicity towards MG-63 osteoblast-like cells. Mentioned features of the obtained Ce and Ga-doped MBGNPs make them useful for multifunctional applications such as drug delivery carriers or bioactive fillers for bone tissue engineering applications.
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Cerio , Galio , Nanopartículas , Antibacterianos/farmacología , Regeneración Ósea , Escherichia coli , Galio/farmacología , Vidrio , Dióxido de Silicio , Staphylococcus aureusRESUMEN
Silicone resins, filled with phosphates and other oxide fillers, yield upon firing in air at 1100 °C, a product resembling Biosilicate® glass-ceramics, one of the most promising systems for tissue engineering applications. The process requires no preliminary synthesis of parent glass, and the polymer route enables the application of direct ink writing (DIW) of silicone-based mixtures, for the manufacturing of reticulated scaffolds at room temperature. The thermal treatment is later applied for the conversion into ceramic scaffolds. The present paper further elucidates the flexibility of the approach. Changes in the reference silicone and firing atmosphere (from air to nitrogen) were studied to obtain functional composite biomaterials featuring a carbon phase embedded in a Biosilicate®-like matrix. The microstructure was further modified either through a controlled gas release at a low temperature, or by the revision of the adopted additive manufacturing technology (from DIW to digital light processing).
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BACKGROUND: Yttria-stabilized zirconia nanoparticles can be applied as fillers to improve the mechanical and antibacterial properties of luting cement. The aim of this study was to synthesize yttria-stabilized zirconia nanoparticles by the sol-gel method and to investigate their composition, structure, morphology and biological properties. METHODS: Nanopowders of ZrO2 7 wt% Y2O3 (nY-ZrO) were synthesized by the sol-gel method and were sintered at three different temperatures: 800, 1000 and 1200 °C, and their composition, size and morphology were investigated. The biocompatibility was investigated with human gingival fibroblasts (hGFs), while reactive oxygen species (ROS) production was evaluated through fluorescence analysis. RESULTS: All synthesized materials were composed of tetragonal zirconia, while nanopowders sintered at 800 °C and 1000 °C additionally contained 5 and 20 wt% of the cubic phase. By increasing the calcination temperature, the crystalline size of the nanoparticles increased from 12.1 nm for nY-ZrO800 to 47.2 nm for nY-ZrO1200. Nano-sized particles with good dispersion and low agglomeration were received. Cell culture studies with human gingival fibroblasts verified the nanopowders' biocompatibility and their ROS scavenging activity. CONCLUSIONS: the obtained sol-gel derived nanopowders showed suitable properties to be potentially used as nanofillers for dental luting cement.
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Chitosan fibers blended with polyethylene oxide (CHIT_PEO) and crosslinked with genipin were fabricated by electrospinning technique. Subsequently, CHIT_PEO bioactive glass composite electrospun mats were fabricated with the aim to achieve flexible structures with adequate mechanical properties and improved biological performance respect to CHIT_PEO fibers, for potential applications in wound healing. Three different compositions of bioactive glasses (BG) were selected and investigated: 45S5 BG, a Sr and Mg containing bioactive glass (BGMS10) and a Zn-containing bioactive glass (BGMS_2Zn). Particulate BGs (particles size < 20 µm) were separately added to the starting CHIT_PEO solution before electrospinning. The two recently developed bioactive glasses (BGMS10 and BGMS_2Zn) showed very promising biological properties in terms of bioactivity and cellular viability; thus, such compositions were added for the first time to CHIT_PEO solution to fabricate composite electrospun mats. The incorporation of bioactive glass particles and their distribution into CHIT_PEO fibers were assessed by SEM and FTIR analyses. Furthermore, CHIT_PEO composite electrospun mats showed improved mechanical properties in terms of Young's Modulus compared to neat CHIT_PEO fibers; on the contrary, the values of tensile strain at break (%) were comparable. Biological performance in terms of cellular viability was investigated by means of WST-8 assay and CHIT_PEO composite electrospun mats showed cytocompatibility and the desired cellular viability.
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Inducing the formation of new blood vessels (angiogenesis) is an essential requirement for successful tissue engineering. Approaches have been proposed to enhance angiogenesis using growth factors and other biomolecules; however, this approaches present drawbacks in terms of high cost and patient safety. Copper is known to effectively regulate angiogenesis and can offer a more cost-effective alternative than the direct use of growth factors. With this study, a strategy to incorporate copper in electrospun fibrous scaffolds with pro-angiogenic properties is presented. Polycaprolactone (PCL) and copper(II)-chitosan are electrospun using benign solvents. The morphological and physicochemical properties of the fiber mats are investigated through scanning electron microscopy (SEM), static contact angle measurements, energy dispersive X-ray, and Fourier-transform infrared spectroscopies. Scaffold stability in phosphate buffered saline at 37 °C is monitored over 1 week. A bone marrow stromal cell line (ST-2) is cultured for 7 days and its behavior is evaluated using SEM, fluorescence microscopy and a tetrazolium salt-based colorimetric assay. Results confirm that PCL/copper(II)-chitosan is suitable for electrospinning. The fiber mats are biocompatible and favor cell colonization and infiltration. Most notably, the angiogenic potential of PCL/copper(II)-chitosan blends is confirmed by a three-fold increase in VEGF secretion by ST-2 cells in the presence of copper(II)-chitosan.