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1.
ACS Mater Au ; 3(6): 636-645, 2023 Nov 08.
Artículo en Inglés | MEDLINE | ID: mdl-38089667

RESUMEN

In recent years, fiber-based systems have been explored in the frame of tissue engineering due to their robustness in recapitulating the architecture and mechanical properties of native tissues. Such scaffolds offer anisotropic architecture capable of reproducing the native collagen fibers' orientation and distribution. Moreover, fibrous constructs might provide a biomimetic environment for cell encapsulation and proliferation as well as influence their orientation and distribution. In this work, we combine two fiber fabrication techniques, such as electrospinning and wet-spinning, in order to obtain novel cell-laden 3D fibrous layered scaffolds which can simultaneously provide: (i) mechanical support; (ii) suitable microenvironment for 3D cell encapsulation; and (iii) loading and sustained release of growth factors for promoting the differentiation of human bone marrow-derived mesenchymal stem cells (hB-MSCs). The constructs are formed from wet-spun hydrogel fibers loaded with hB-MSCs deposited on a fibrous composite electrospun matrix made of polycaprolactone, polyamide 6, and mesoporous silica nanoparticles enriched with bone morphogenetic protein-12 (BMP-12). Morphological and mechanical characterizations of the structures were carried out, and the growth factor release was assessed. The biological response in terms of cell viability, alignment, differentiation, and extracellular matrix production was investigated. Ex vivo testing of the layered structure was performed to prove the layers' integrity when subjected to mechanical stretching in the physiological range. The results reveal that 3D layered scaffolds can be proposed as valid candidates for tendon tissue engineering.

2.
ACS Omega ; 8(31): 28109-28121, 2023 Aug 08.
Artículo en Inglés | MEDLINE | ID: mdl-37576652

RESUMEN

In this study, two-layer poly(vinyl alcohol)/gelatin (PVA/GEL) nanofiber patches containing cinnamaldehyde (CA) in the first layer and gentamicin (GEN) in the second layer were produced by the electrospinning method. The morphology, chemical structures, and thermal temperatures of the produced pure (PVA/GEL), CA-loaded (PVA/GEL/CA), GEN-loaded (PVA/GEL/GEN), and combined drug-loaded (PVA/GEL/CA/GEN) nanofiber patches were determined by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy, and differential scanning calorimetry, respectively. Their mechanical properties, swelling and degradation behavior, and drug release kinetics were investigated. SEM images showed that both drug-free and drug-loaded nanofiber patches possess smooth and monodisperse structures, and nanofiber size increase occurred as the amount of drug increased. The tensile test results showed that the mechanical strength decreased as the drug was loaded. According to the drug release results, CA release ended at the 96th hour, while GEN release continued until the 264th hour. The antibacterial and antibiofilm activities of PVA/GEL, PVA/GEL/CA, PVA/GEL/GEN, and PVA/GEL/CA/GEN nanofiber patches against Pseudomonas aeruginosa and Staphylococcus aureus were evaluated. Results showed that PVA/GEL/GEN and PVA/GEL/CA/GEN nanofiber patches have excellent antibacterial and antibiofilm activities. Moreover, all materials were biocompatible, with no cytotoxic effects in the mammalian cell model for 8 days. PVA/GEL/GEN nanofiber patches were the most promising material for a high cell survival ratio, which was confirmed by SEM images. This research aims to develop an alternative method to stop and treat the rapid progression of bacterial keratitis.

3.
Polymers (Basel) ; 14(24)2022 Dec 13.
Artículo en Inglés | MEDLINE | ID: mdl-36559835

RESUMEN

Fibrous shape memory scaffolds composed of thermoplastic polyurethane based on a mixture of polycaprolactone diols were fabricated. The effect of the fiber diameter and arrangement- random (rPU) or aligned (aPU), on crystallinity, mechanical properties, and shape memory was analyzed. The diameters of the fibers were controlled by changing the concentration of polyurethane (PU) solutions in the range of 5% to 16% and fibers alignment by utilization of different collectors. The chemical structure was confirmed by Fourier Transformed Infrared spectroscopy (FTIR), crystallinity was evaluated based on differential scanning calorimetry (DSC,) and mechanical properties were measured by the tensile test. Additionally, shape memory programming was performed using a dynamic mechanical analyzer (DMA), and shape recovery was evaluated in the air and in the water environment. DSC results showed that the electrospinning process did not change the crystallinity or melting temperature of synthesized thermoplastic polyurethanes. The melting temperature of the crystalline switching segments was around 26-27 °C, and the crystalline phase of hard segments was around 130 °C. Shape memory properties were analyzed in the contest of the fiber diameter and alignment of the fibers, while changes in the fibers' diameters from 360 nm to 1760 nm did not result in significant changes in shape recovery coefficient (Rr) especially evaluated in the air. The longitudinal fiber alignment enhanced mechanical and shape recovery to up to 96% for aPU, with the highest fiber diameter when evaluated in water.

4.
Tissue Cell ; 78: 101899, 2022 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-36030673

RESUMEN

BACKGROUND: The development of tissue-engineered scaffolds with electrical properties is the primary motivation of novel regenerative medicine. Electroconductive scaffolds are designed to mimic the injured tissue environment's electrical properties and regulate cellular behavior - growth, proliferation, and differentiation - that could stimulate the injured nerve's regeneration. METHODS: We fabricated dedicated electroconductive scaffolds and customized an appropriate device with an external current supply to expose cells on the scaffold to electrical stimulation (ES). Next, we isolated rat adipose-derived stem cells (ASCs) and performed in vitro experiments that combine cells, an electroconductive scaffold, NGF (nerve growth factor), and ES (90 mV/mm, constant, for four days). Finally, we checked cellular activity as proliferation, viability, morphology, the neurogenic differentiation potential of ASCs, cell alignment, and karyotype. RESULTS: We observed that the electrical stimulation did not change the viability and chromosome stability of rat ASCs, but altered slightly proliferation compared to non-stimulated cells. The combined effect of a scaffold, NGF, and ES caused morphology changes and enhancement of ASCs neuronal differentiation as indicated in ßIII-tubulin expression, actin organization, and upregulation of neurogenic gene expression. CONCLUSIONS: We developed an electroconductive scaffold and customized device for in vitro study with many experimental variants. Based on our results, we presumed that the established study scheme - including an electroconductive scaffold, NGF and ES - is biocompatible and could guide ASCs to differentiate in neurogenic lineage, thus may be potentially applied in nerve injury regeneration.


Asunto(s)
Células Madre Mesenquimatosas , Nanofibras , Actinas/metabolismo , Tejido Adiposo , Animales , Diferenciación Celular , Factor de Crecimiento Nervioso/metabolismo , Factor de Crecimiento Nervioso/farmacología , Ratas , Andamios del Tejido , Tubulina (Proteína)
5.
Polymers (Basel) ; 14(14)2022 Jul 16.
Artículo en Inglés | MEDLINE | ID: mdl-35890657

RESUMEN

Plastic materials are one of the significant components of construction materials omnipresent in all areas of the industry and everyday life. One of these plastics is polyethylene terephthalate (PET). Due to its processing properties, with a simultaneous low production cost, PET has been used in many industrial applications, including the production of various types of bottles. Moreover, the high consumption of PET bottles causes the accumulation of large amounts of their waste and necessitates finding an effective way to recycle them. Electrospinning is a well-known non-complicated method for the fabrication of nonwovens from polymers and composites, which can be utilized in many fields due to their outstanding properties. In addition, it might be a promising technique for the recycling of plastic materials. Therefore, in this study, the electrospinning approach for the recycling of two types of PET bottle wastes-bottles made of virgin PET and bottles made of recycled PET (PET bottles) has been utilized, and a comparison of the properties of the obtained materials have been performed. The fibers with diameters of 1.62 ± 0.22, 1.64 ± 0.18, and 1.89 ± 0.19 have been produced from solutions made of virgin PET granulate, PET bottles, and PET bottles made of recycled bottles, respectively. Obtained fibers underwent morphological observation using a scanning electron microscope. Physico-chemical properties using FTIR, gel chromatography, and differential scanning calorimetry have been evaluated, and mechanical properties of obtained mats have been investigated. Cytotoxicity tests using the L929 mouse fibroblast cell line revealed no cytotoxicity for all tested materials.

6.
Int J Mol Sci ; 22(24)2021 Dec 08.
Artículo en Inglés | MEDLINE | ID: mdl-34948005

RESUMEN

In this work, NiTi alloy parts were fabricated using laser powder bed fusion (LBPF) from pre-alloyed NiTi powder and in situ alloyed pure Ni and Ti powders. Comparative research on the corrosive and biological properties of both studied materials was performed. Electrochemical corrosion tests were carried out in phosphate buffered saline at 37 °C, and the degradation rate of the materials was described based on Ni ion release measurements. Cytotoxicity, bacterial growth, and adhesion to the surface of the fabricated coupons were evaluated using L929 cells and spherical Escherichia coli (E. coli) bacteria, respectively. The in situ alloyed NiTi parts exhibit slightly lower corrosion resistance in phosphate buffered saline solution than pre-alloyed NiTi. Moreover, the passive layer formed on in situ alloyed NiTi is weaker than the one formed on the NiTi fabricated from pre-alloyed NiTi powder. Furthermore, in situ alloyed NiTi and NiTi made from pre-alloyed powders have comparable cytotoxicity and biological properties. Overall, the research has shown that nitinol sintered using in situ alloyed pure Ni and Ti is potentially useful for biomedical applications.


Asunto(s)
Aleaciones/farmacología , Escherichia coli/fisiología , Níquel/química , Titanio/química , Aleaciones/química , Animales , Adhesión Bacteriana/efectos de los fármacos , Línea Celular , Corrosión , Escherichia coli/efectos de los fármacos , Ensayo de Materiales , Ratones , Fosfatos/química , Polvos , Propiedades de Superficie
7.
Biosensors (Basel) ; 11(7)2021 Jul 08.
Artículo en Inglés | MEDLINE | ID: mdl-34356698

RESUMEN

This research presents an electrochemical immunosensor for collagen I detection using a self-assembled monolayer (SAM) of gold nanoparticles (AuNPs) and covalently immobilized half-reduced monoclonal antibody as a receptor; this allowed for the validation of the collagen I concentration through two different independent methods: electrochemically by Electrochemical Impedance Spectroscopy (EIS), and optically by Surface Plasmon Resonance (SPR). The high unique advantage of the proposed sensor is based on the performance of the stable covalent immobilization of the AuNPs and enzymatically reduced half-IgG collagen I antibodies, which ensured their appropriate orientation onto the sensor's surface, good stability, and sensitivity properties. The detection of collagen type I was performed in a concentration range from 1 to 5 pg/mL. Moreover, SPR was utilized to confirm the immobilization of the monoclonal half-antibodies and sensing of collagen I versus time. Furthermore, EIS experiments revealed a limit of detection (LOD) of 0.38 pg/mL. The selectivity of the performed immunosensor was confirmed by negligible responses for BSA. The performed approach of the immunosensor is a novel, innovative attempt that enables the detection of collagen I with very high sensitivity in the range of pg/mL, which is significantly lower than the commonly used enzyme-linked immunosorbent assay (ELISA).


Asunto(s)
Espectroscopía Dieléctrica , Inmunoensayo , Resonancia por Plasmón de Superficie , Anticuerpos Inmovilizados , Anticuerpos Monoclonales , Técnicas Biosensibles , Colágeno Tipo I , Electrodos , Ensayo de Inmunoadsorción Enzimática , Oro/química , Límite de Detección , Nanopartículas del Metal
8.
Materials (Basel) ; 14(15)2021 Jul 23.
Artículo en Inglés | MEDLINE | ID: mdl-34361306

RESUMEN

Graphene and its derivatives have attracted scientists' interest due to their exceptional properties, making them alluring candidates for multiple applications. However, still little is known about the properties of as-obtained graphene derivatives during long-term storage. The aim of this study was to check whether or not 14 months of storage time impacts graphene oxide flakes' suspension purity. Complementary micro and nanoscale characterization techniques (SEM, AFM, EDS, FTIR, Raman spectroscopy, and elemental combustion analysis) were implemented for a detailed description of the topography and chemical properties of graphene oxide flakes. The final step was pH evaluation of as-obtained and aged samples. Our findings show that purified flakes sustained their purity over 14 months of storage.

9.
Int J Mol Sci ; 22(11)2021 May 25.
Artículo en Inglés | MEDLINE | ID: mdl-34070436

RESUMEN

The aim was to examine the efficiency of a scaffold made of poly (L-lactic acid)-co-poly(ϵ-caprolactone), collagen (COL), polyaniline (PANI), and enriched with adipose-derived stem cells (ASCs) as a nerve conduit in a rat model. P(LLA-CL)-COL-PANI scaffold was optimized and electrospun into a tubular-shaped structure. Adipose tissue from 10 Lewis rats was harvested for ASCs culture. A total of 28 inbred male Lewis rats underwent sciatic nerve transection and excision of a 10 mm nerve trunk fragment. In Group A, the nerve gap remained untouched; in Group B, an excised trunk was used as an autograft; in Group C, nerve stumps were secured with P(LLA-CL)-COL-PANI conduit; in Group D, P(LLA-CL)-COL-PANI conduit was enriched with ASCs. After 6 months of observation, rats were sacrificed. Gastrocnemius muscles and sciatic nerves were harvested for weight, histology analysis, and nerve fiber count analyses. Group A showed advanced atrophy of the muscle, and each intervention (B, C, D) prevented muscle mass decrease (p < 0.0001); however, ASCs addition decreased efficiency vs. autograft (p < 0.05). Nerve fiber count revealed a superior effect in the nerve fiber density observed in the groups with the use of conduit (D vs. B p < 0.0001, C vs. B p < 0.001). P(LLA-CL)-COL-PANI conduits with ASCs showed promising results in managing nerve gap by decreasing muscle atrophy.


Asunto(s)
Modelos Animales de Enfermedad , Células Madre Mesenquimatosas/metabolismo , Nanofibras/química , Regeneración Nerviosa , Neurogénesis , Traumatismos de los Nervios Periféricos/terapia , Nervio Ciático/metabolismo , Andamios del Tejido/química , Compuestos de Anilina/química , Animales , Caproatos/química , Células Cultivadas , Colágeno/química , Inmunohistoquímica , Lactonas/química , Masculino , Ensayo de Materiales , Células Madre Mesenquimatosas/citología , Microscopía Electrónica de Rastreo , Músculo Esquelético/citología , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Nanofibras/ultraestructura , Traumatismos de los Nervios Periféricos/metabolismo , Traumatismos de los Nervios Periféricos/patología , Poliésteres/química , Ratas , Ratas Endogámicas Lew , Nervio Ciático/citología , Nervio Ciático/patología , Trasplante Autólogo
10.
Adv Healthc Mater ; 10(7): e2001305, 2021 04.
Artículo en Inglés | MEDLINE | ID: mdl-33576158

RESUMEN

Tendon and ligament injuries caused by trauma and degenerative diseases are frequent and affect diverse groups of the population. Such injuries reduce musculoskeletal performance, limit joint mobility, and lower people's comfort. Currently, various treatment strategies and surgical procedures are used to heal, repair, and restore the native tissue function. However, these strategies are inadequate and, in some cases, fail to re-establish the lost functionality. Tissue engineering and regenerative medicine approaches aim to overcome these disadvantages by stimulating the regeneration and formation of neotissues. Design and fabrication of artificial scaffolds with tailored mechanical properties are crucial for restoring the mechanical function of tendons. In this review, the tendon and ligament structure, their physiology, and performance are presented. On the other hand, the requirements are focused for the development of an effective reconstruction device. The most common fiber-based scaffolding systems are also described for tendon and ligament tissue regeneration like strand fibers, woven, knitted, braided, and braid-twisted fibrous structures, as well as electrospun and wet-spun constructs, discussing critically the advantages and limitations of their utilization. Finally, the potential of multilayered systems as the most effective candidates for tendon and ligaments tissue engineering is pointed out.


Asunto(s)
Tendones , Andamios del Tejido , Humanos , Ligamentos , Medicina Regenerativa , Ingeniería de Tejidos
11.
Int J Biol Macromol ; 165(Pt B): 2049-2059, 2020 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-33086111

RESUMEN

Composite polycaprolactone-chitosan material was produced by an electrospinning method and used as a support for immobilization of tyrosinase by mixed ionic interactions and hydrogen bonds formation. The morphology of the fibers and enzyme deposition were confirmed by SEM images. Further, multivariate polynomial regression was used to model the experimental data and to determine optimal conditions for immobilization process, which were found to be pH 7, temperature 25 °C and 16 h process duration. Under these conditions, novel type of biocatalytic system was produced with immobilization yield of 93% and expressed activity of 95%. Furthermore, as prepared system was applied in batch experiments related to biodegradation of bisphenol A under various remediation conditions. It was found that over 80% of the pollutant was removed after 120 min of the process, in the temperature range 15-45 °C and pH 6-9, using solutions at concentration up to 3 mg/L. Experimental data collected proved that the stability and reusability of the tyrosinase were significantly improved upon immobilization: the immobilized biomolecule retained around 90% of its initial activity after 30 days of storage, and was still capable to remove over 80% of bisphenol A even after 10 repeated uses. By contrast, free enzyme was able to remove over 80% of bisphenol A at pH 7-8 and temperature range 15-35 °C, and retained less than 60% of its initial activity after 30 days of storage.


Asunto(s)
Compuestos de Bencidrilo/aislamiento & purificación , Quitosano/química , Enzimas Inmovilizadas/metabolismo , Monofenol Monooxigenasa/metabolismo , Fenoles/aislamiento & purificación , Poliésteres/química , Agaricales/enzimología , Biodegradación Ambiental , Enzimas Inmovilizadas/ultraestructura , Concentración de Iones de Hidrógeno , Monofenol Monooxigenasa/ultraestructura , Espectroscopía Infrarroja por Transformada de Fourier
12.
Mater Sci Eng C Mater Biol Appl ; 110: 110675, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32204102

RESUMEN

Integration of multiple features including shape memory, biodegradation, and sustained drug delivery in a single material offers the opportunity to significantly improve the abilities of implantable devices for cardiovascular system regeneration. Two types of shape memory polyurethanes (SMPUs): PU-PLGA and PU-PLLA/PEG differing in soft segments composition that comprising blends of various biodegradable polyols, i.e. D,l-lactide-co-glycolide diol (o-PLGA), poly(e-caprolactone) diols (o-PCL) with various molecular weights, poly-l-lactide diol (o-PLLA), polyethylene glycol (o-PEG) were synthesized and further utilized to electrospun nanofibrous - rapamycin (Rap) delivery system. Structure characterization by Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DCS) and hydrophilicity measurements were performed to gain more insights on the influence of the particular units of the softs segments on the transition temperature (Ttrans), shape recovery, degradation profile, and drug release kinetics. In vitro study in PBS solution revealed that incorporation of o-PLGA segments to SMPUs is favorable over o-PEG as increased shape memory performance was observed. Moreover, presence of PLGA in PU-PLGA gave more predictable degradation profile in comparison to PU-PLLA/PEG system. Human Cardiac Fibroblasts (HCF) viability tests in vitro confirmed that the amount of Rap released from evaluated PU-PLLA/PEG/Rap and PU-PLGA/Rap drug delivery systems was sufficient to inhibit cells growth on the surface of the tested materials.


Asunto(s)
Sistemas de Liberación de Medicamentos , Poliuretanos/química , Materiales Inteligentes/química , Ingeniería de Tejidos , Materiales Biocompatibles/química , Rastreo Diferencial de Calorimetría , Supervivencia Celular , Cristalización , Liberación de Fármacos , Fibroblastos/citología , Humanos , Cinética , Lactatos , Peso Molecular , Poliésteres , Polietilenglicoles , Espectroscopía Infrarroja por Transformada de Fourier , Temperatura , Resistencia a la Tracción
13.
Environ Res ; 184: 109332, 2020 05.
Artículo en Inglés | MEDLINE | ID: mdl-32151845

RESUMEN

Novel electrospun poly(methyl methacrylate)/polyaniline electrospun fibres were produced, characterised, modified, and used as a support for laccase immobilisation by two methods: adsorption and covalent binding. Effective deposition of laccase by both methods was confirmed by FTIR and CLSM results. Nevertheless, the main objective of the study was to select the most favourable immobilisation conditions and prepare heterogeneous biocatalysts with the best possible catalytic properties. The highest relative activity of enzymes immobilised by adsorption and covalent binding were obtained after 1 h of immobilisation using laccase solution at a concentration of 1 mg/mL, at pH 5 and 25 °C. It was found that the immobilised enzymes, which were present in amounts of 110 mg/g and 185 mg/g for systems with adsorbed and covalently bonded laccase respectively, exhibited slightly lower substrate affinity, and in consequence also a lower maximum reaction rate, than the free enzyme. The stability of laccase improved significantly upon immobilisation: both heterogeneous biocatalysts retained over 80% relative activity even after 10 repeated catalytic cycles and 30 days of storage. The obtained systems were used for decolourisation of Remazol Brilliant Blue R dye from a model aqueous solution, resulting in removal efficiencies of 87% and 58% using adsorbed and covalently bonded laccase, respectively. The described approach to the removal of textile dye from model solution is significant for the sustainable and environmentally friendly decolourisation of various compounds from wastewater.


Asunto(s)
Colorantes , Lacasa , Polimetil Metacrilato , Purificación del Agua , Adsorción , Compuestos de Anilina
14.
Polymers (Basel) ; 12(2)2020 Feb 02.
Artículo en Inglés | MEDLINE | ID: mdl-32024273

RESUMEN

Poly(2-oxazoline) (POx) matrices in the form of non-woven fibrous mats and three-dimensional moulds were obtained by electrospinning and fused deposition modelling (FDM), respectively. To obtain these materials, poly(2-isopropyl-2-oxazoline) (PiPrOx) and gradient copolymers of 2-isopropyl- with 2-n-propyl-2-oxazoline (P(iPrOx-nPrOx)), with relatively low molar masses and low dispersity values, were processed. The conditions for the electrospinning of POx were optimised for both water and the organic solvent. Also, the FDM conditions for the fabrication of POx multi-layer moulds of cylindrical or cubical shape were optimised. The properties of the POx after electrospinning and extrusion from melt were determined. The molar mass of all (co)poly(2-oxazoline)s did not change after electrospinning. Also, FDM did not influence the molar masses of the (co)polymers; however, the long processing of the material caused degradation and an increase in molar mass dispersity. The thermal properties changed significantly after processing of POx what was monitored by increase in enthalpy of exo- and endothermic peaks in differential scanning calorimetry (DSC) curve. The influence of the processing conditions on the structure and properties of the final material were evaluated having in a mind their potential application as scaffolds.

15.
J Mech Behav Biomed Mater ; 101: 103437, 2020 01.
Artículo en Inglés | MEDLINE | ID: mdl-31557662

RESUMEN

Biodegradable electrospun nanofibrous scaffolds for bone tissue engineering applications have been extensively studied as they can provide attractive open-worked architecture resembling natural extracellular matrix, with tunable physical and mechanical properties enhancing positive cellular response. For this purpose, electrospun mats were tested in terms of morphology, mechanical and physical properties, degradation kinetics and related phenomena occurring in micro- and nanoscale. However, detailed description of internal nanostructures of electrospun mats and their changes related to in vitro degradation is still missing. In this manuscript, we report qualitative and quantitative evaluation of internal lamellar nanostructure of electrospun fibrous scaffolds made of pristine polycaprolactone and composite with polymeric matrix and nanoceramic (hydroxyapatite) filler during in vitro degradation. Morphological and mechanical studies performed with an atomic force microscope were followed by scanning electron microscope imaging and X-Ray diffraction. The results suggest degradation-dependent alteration of both organization and thickness of nano-scaled lamellas recorded with atomic force microscope. Moreover, changes of the material's internal structure were followed by enhanced stiffness and higher crystallinity of electrospun fibers.


Asunto(s)
Electricidad , Fenómenos Mecánicos , Microscopía de Fuerza Atómica , Nanopartículas/química , Poliésteres/química , Materiales Biocompatibles/química , Materiales Biocompatibles/metabolismo , Durapatita/química , Cinética , Nanoestructuras/química , Poliésteres/metabolismo
16.
Mater Sci Eng C Mater Biol Appl ; 103: 109789, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31349507

RESUMEN

Enzymatic biodegradation of pharmaceuticals, using enzymes such as laccase, is a green solution for the removal of toxic pollutants that has attracted growing interest over recent years. Moreover, the application of immobilized biocatalysts is relevant for industrial applications, due to the improved stability and reusability of the immobilized enzymes. Thus, in the present study, laccase was immobilized by adsorption and encapsulation using poly(l-lactic acid)-co-poly(ε-caprolactone) (PLCL) electrospun nanofibers as a tailor-made support. The produced biocatalytic systems were applied in the biodegradation of two commonly used anti-inflammatories, naproxen and diclofenac, which are present in wastewaters at environmentally relevant concentrations. The results showed that under optimal process conditions (temperature 25 °C, pH 5 and 3 for naproxen and diclofenac respectively), even from a solution at a concentration of 1 mg L-1, over 90% of both pharmaceuticals was removed by encapsulated laccase in batch mode. Both immobilized enzymes also exhibited high reusability: after five reaction cycles approximately 60% and 40% of naproxen and diclofenac were removed by encapsulated and adsorbed laccase respectively. In addition, a thorough analysis was made of the products of biodegradation of the two studied pollutants. Furthermore, toxicity study of the mixture after biodegradation of the pharmaceuticals showed that the solutions obtained after the process were approximately 65% less toxic than the initial naproxen and diclofenac solutions.


Asunto(s)
Biodegradación Ambiental , Diclofenaco/metabolismo , Lacasa/metabolismo , Nanofibras/química , Naproxeno/metabolismo , Animales , Artemia/efectos de los fármacos , Biocatálisis , Diclofenaco/química , Estabilidad de Enzimas , Enzimas Inmovilizadas/metabolismo , Equipo Reutilizado , Concentración de Iones de Hidrógeno , Naproxeno/química , Poliésteres/química , Temperatura , Contaminantes Químicos del Agua/química , Contaminantes Químicos del Agua/metabolismo , Contaminantes Químicos del Agua/toxicidad
17.
Nanomaterials (Basel) ; 9(3)2019 Mar 05.
Artículo en Inglés | MEDLINE | ID: mdl-30841516

RESUMEN

The detection and monitoring of circulating tumor cells (CTCs) in blood is an important strategy for early cancer evidence, analysis, monitoring of therapeutic response, and optimization of cancer therapy treatments. In this work, tailor-made membranes (MBSP) for surface-enhanced Raman spectroscopy (SERS)-based analysis, which permitted the separation and enrichment of CTCs from blood samples, were developed. A thin layer of SERS-active metals deposited on polymer mat enhanced the Raman signals of CTCs and provided further insight into CTCs molecular and biochemical composition. The SERS spectra of all studied cells-prostate cancer (PC3), cervical carcinoma (HeLa), and leucocytes as an example of healthy (normal) cell-revealed significant differences in both the band positions and/or their relative intensities. The multivariate statistical technique based on principal component analysis (PCA) was applied to identify the most significant differences (marker bands) in SERS data among the analyzed cells and to perform quantitative analysis of SERS data. Based on a developed PCA algorithm, the studied cell types were classified with an accuracy of 95% in 2D PCA to 98% in 3D PCA. These results clearly indicate the diagnostic efficiency for the discrimination between cancer and normal cells. In our approach, we exploited the one-step technology that exceeds most of the multi-stage CTCs analysis methods used and enables simultaneous filtration, enrichment, and identification of the tumor cells from blood specimens.

18.
Adv Healthc Mater ; 8(7): e1801218, 2019 04.
Artículo en Inglés | MEDLINE | ID: mdl-30725521

RESUMEN

Fiber-based approaches hold great promise for tendon tissue engineering enabling the possibility of manufacturing aligned hydrogel filaments that can guide collagen fiber orientation, thereby providing a biomimetic micro-environment for cell attachment, orientation, migration, and proliferation. In this study, a 3D system composed of cell-laden, highly aligned hydrogel yarns is designed and obtained via wet spinning in order to reproduce the morphology and structure of tendon fascicles. A bioink composed of alginate and gelatin methacryloyl (GelMA) is optimized for spinning and loaded with human bone morrow mesenchymal stem cells (hBM-MSCs). The produced scaffolds are subjected to mechanical stretching to recapitulate the strains occurring in native tendon tissue. Stem cell differentiation is promoted by addition of bone morphogenetic protein 12 (BMP-12) in the culture medium. The aligned orientation of the fibers combined with mechanical stimulation results in highly preferential longitudinal cell orientation and demonstrates enhanced collagen type I and III expression. Additionally, the combination of biochemical and mechanical stimulations promotes the expression of specific tenogenic markers, signatures of efficient cell differentiation towards tendon. The obtained results suggest that the proposed 3D cell-laden aligned system can be used for engineering of scaffolds for tendon regeneration.


Asunto(s)
Hidrogeles/química , Estrés Mecánico , Tendones/citología , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Alginatos/química , Materiales Biocompatibles/química , Proteínas Morfogenéticas Óseas/química , Proteínas Morfogenéticas Óseas/farmacología , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Colágeno Tipo III/genética , Colágeno Tipo III/metabolismo , Gelatina/química , Humanos , Tinta , Dispositivos Laboratorio en un Chip , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Impresión Tridimensional , Tendones/metabolismo , Ingeniería de Tejidos/instrumentación
19.
Mater Sci Eng C Mater Biol Appl ; 94: 516-523, 2019 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-30423736

RESUMEN

Bioactive glass-based scaffolds are commonly used in bone tissue engineering due to their biocompatibility, mechanical strength and adequate porous structure. However, their hydrophobicity and brittleness limits their practical application. In this study, to improve nanomechanical properties of such scaffolds, pure bioactive hybrid glass and two bioactive hybrid glass-polymer coated composites were fabricated. A complementary micro and nanoscale characterization techniques (SEM, AFM, µCT, FTIR, compressive test, goniometer) were implemented for detailed description of architecture and physicochemical properties of hybrid bioactive glass-based scaffolds with emphasis on nano-mechanics. The final step was in-vitro evaluation of three dimensional macroporous structures. Our findings show that after polymer addition, architecture, topography and surface properties of the scaffolds were changed and promoted favoured behaviour of the cells.


Asunto(s)
Huesos/fisiología , Cerámica/química , Materiales Biocompatibles Revestidos/química , Polímeros/química , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Línea Celular Tumoral , Supervivencia Celular , Módulo de Elasticidad , Humanos , Nanopartículas/química , Porosidad , Espectroscopía Infrarroja por Transformada de Fourier , Estrés Mecánico , Microtomografía por Rayos X
20.
ACS Biomater Sci Eng ; 5(6): 2953-2964, 2019 Jun 10.
Artículo en Inglés | MEDLINE | ID: mdl-33405598

RESUMEN

Tendon injuries are frequent and occur in the elderly, young, and athletic populations. The inadequate number of donors combined with many challenges associated with autografts, allografts, xenografts, and prosthetic devices have added to the value of engineering biological substitutes, which can be implanted to repair the damaged tendons. Electrospun scaffolds have the potential to mimic the native tissue structure along with desired mechanical properties and, thus, have attracted noticeable attention. In order to improve the biological responses of these fibrous structures, we designed and fabricated 3D multilayered composite scaffolds, where an electrospun nanofibrous substrate was coated with a thin layer of cell-laden hydrogel. The whole construct composition was optimized to achieve adequate mechanical and physical properties as well as cell viability and proliferation. Mesenchymal stem cells (MSCs) were differentiated by the addition of bone morphogenetic protein 12 (BMP-12). To mimic the natural function of tendons, the cell-laden scaffolds were mechanically stimulated using a custom-built bioreactor. The synergistic effect of mechanical and biochemical stimulation was observed in terms of enhanced cell viability, proliferation, alignment, and tenogenic differentiation. The results suggested that the proposed constructs can be used for engineering functional tendons.

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