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To improve the capability of non-woven polypropylene-based fabric (NWF-PP) used for face mask production to retain active biomolecules such as polyphenols, the surface functionalization of NWF-PP-directly cut from face masks-was carried out by employing cold plasma with oxygen. The nature/structure of the functional groups, as well as the degree of functionalization, were evaluated by ATR-FTIR and XPS by varying the experimental conditions (generator power, treatment time, and oxygen flow). The effects of plasma activation on mechanical and morphological characteristics were evaluated by stress-strain measurements and SEM analysis. The ability of functionalized NWF-PP to firmly anchor polyphenols extracted from cloves was estimated by ATR-FTIR analysis, IR imaging, extractions in physiological solution, and OIT analysis (before and after extraction), as well as by SEM analysis. All the results obtained converge in showing that, although the plasma treatment causes changes-not only on the surface-with certain detriment to the mechanical performance of the NWF-PP, the incorporated functionalities are able to retain/anchor the active molecules extracted from the cloves, thus stabilizing the treated surfaces against thermo-oxidation even after prolonged extraction.
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Gases em Plasma , Polifenóis , Polipropilenos/química , OxigênioRESUMO
Polyhydroxyalkanoates (PHAs) are a family of biopolyesters synthesized by various microorganisms. Due to their biocompatibility and biodegradation, PHAs have been proposed for biomedical applications, including tissue engineering scaffolds. Olive leaf extract (OLE) can be obtained from agri-food biowaste and is a source of polyphenols with remarkable antioxidant properties. This study aimed at incorporating OLE inside poly(hydroxybutyrate-co-hydroxyvalerate) (PHBHV) fibers via electrospinning to obtain bioactive bio-based blends that are useful in wound healing. PHBHV/OLE electrospun fibers with a size of 1.29 ± 0.34 µm were obtained. Fourier transform infrared chemical analysis showed a uniform surface distribution of hydrophilic -OH groups, confirming the presence of OLE in the electrospun fibers. The main OLE phenols were released from the fibers within 6 days. The biodegradation of the scaffolds in phosphate buffered saline was investigated, demonstrating an adequate stability in the presence of metalloproteinase 9 (MMP-9), an enzyme produced in chronic wounds. The scaffolds were preliminarily tested in vitro with HFFF2 fibroblasts and HaCaT keratinocytes, suggesting adequate cytocompatibility. PHBHV/OLE fiber meshes hold promising features for wound healing, including the treatment of ulcers, due to the long period of durability in an inflamed tissue environment and adequate cytocompatibility.
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Poli-Hidroxialcanoatos , Antioxidantes/farmacologia , Hidroxibutiratos/farmacologia , Metaloproteinase 9 da Matriz , Olea , Ácidos Pentanoicos , Fosfatos , Extratos Vegetais , Poliésteres/química , Poli-Hidroxialcanoatos/química , Polifenóis , Estudos Prospectivos , Engenharia Tecidual , Alicerces Teciduais/química , CicatrizaçãoRESUMO
3D scaffolds used to repair damaged tissues should be able to mimic both composition and functions of natural extracellular matrix, which is mainly composed of polysaccharides and proteins. In our previous research new biomimetic sponges, based on blends of alginate with gelatin, were produced and characterized for myocardial tissue engineering applications. It was observed that these scaffolds can potentially function as a promising cardiac extracellular matrix substitute, but a reinforcement is required to improve their suturing properties. Aim of the present work was the development of a suturable biomimetic patch by the inclusion of a synthetic mesh within an alginate/gelatin scaffold. The mesh, produced by dry spinning, was made of eight superimposed layers of polycaprolactone microfibers, each one rotated of 45° with respect to the adjacent one. Reinforced scaffolds were obtained through the use of a mold, specially designed to place the fibrous mesh exactly in the center of the sponge. Both the reinforcement mesh and the reinforced scaffold were characterized. A perfect integration between the mesh and the sponge was observed. The fibrous mesh reduced the capacity of the sponge to absorb water, but the degree of hydrophilicity of the material was still comparable with that of natural cardiac tissue. The reinforced system showed a suitable stability in aqueous environment and it resulted much more resistant to suturing than not reinforced scaffold and even than human arteries. Polycaprolactone mesh was not cytotoxic and the reinforced scaffold was able to support cardiomyocytes adhesion and proliferation. Overall, the obtained results confirmed that the choice to modify the alginate/gelatin sponges through the insertion of an appropriate reinforcement system turned out to be correct in view of their potential use in myocardial tissue engineering.
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Alginatos/química , Materiais Biomiméticos/química , Gelatina/química , Alicerces Teciduais , Animais , Adesão Celular , Linhagem Celular , Sobrevivência Celular , Colorimetria , Humanos , Camundongos , Ratos , Engenharia Tecidual/métodosRESUMO
The objective of this study was the preparation and physico-chemical, mechanical, biological, and functional characterization of a multifunctional coating for an innovative, fully implantable device. The multifunctional coating was designed to have three fundamental properties: adhesion to device, close mechanical resemblance to human soft tissues, and control of the inflammatory response and tissue repair process. This aim was fulfilled by preparing a multilayered coating based on three components: a hydrophilic primer to allow device adhesion, a poly(vinyl alcohol) hydrogel layer to provide good mechanical compliance with the human tissue, and a layer of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) fibers. The use of biopolymer fibers offered the potential for a long-term interface able to modulate the release of an anti-inflammatory drug (dexamethasone), thus contrasting acute and chronic inflammation response following device implantation. Two copolymers, poly(vinyl acetate-acrylic acid) and poly(vinyl alcohol-acrylic acid), were synthetized and characterized using thermal analysis (DSC, TGA), Fourier transform infrared spectroscopy (FT-IR chemical imaging), in vitro cell viability, and an adhesion test. The resulting hydrogels were biocompatible, biostable, mechanically compatible with soft tissues, and able to incorporate and release the drug. Finally, the multifunctional coating showed a good adhesion to titanium substrate, no in vitro cytotoxicity, and a prolonged and controlled drug release.
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Materiais Revestidos Biocompatíveis/química , Próteses e Implantes , Fenômenos Químicos , Técnicas de Química Sintética , Humanos , Hidrogéis/química , Fenômenos Mecânicos , TermodinâmicaRESUMO
The poly(butylene succinate-co-adipate) (PBSA) is emerging as environmentally sustainable polyester for applications in marine environment. In this work the capacity of microbiome associated with marine plankton culture to degrade PBSA, was tested. A taxonomic and functional characterization of the microbiome associated with the copepod Acartia tonsa, reared in controlled conditions, was analysed by 16S rDNA metabarcoding, in newly-formed adult stages and after 7 d of incubation. A predictive functional metagenomic profile was inferred for hydrolytic activities involved in bioplastic degradation with a particular focus on PBSA. The copepod-microbiome was also characterized in newly-formed carcasses of A. tonsa, and after 7 and 33 d of incubation in the plankton culture medium. Copepod-microbiome showed hydrolytic activities at all developmental stages of the alive copepods and their carcasses, however, the evenness of the hydrolytic bacterial community significantly increased with the time of incubation in carcasses. Microbial genera, never described in association with copepods: Devosia, Kordia, Lentibacter, Methylotenera, Rheinheimera, Marinagarivorans, Paraglaciecola, Pseudophaeobacter, Gaiella, Streptomyces and Kribbella sps., were retrieved. Kribbella sp. showed carboxylesterase activity and Streptomyces sp. showed carboxylesterase, triacylglycerol lipase and cutinase activities, that might be involved in PBSA degradation. A culturomic approach, adopted to isolate bacterial specimen from carcasses, led to the isolation of the bacterial strain, Vibrio sp. 01 tested for the capacity to promote the hydrolysis of the ester bonds. Granules of PBSA, incubated 82 d at 20 °C with Vibrio sp. 01, were characterized by scanning electron microscopy, infrared spectroscopy, thermogravimetric analysis, and differential scanning calorimetry, showing fractures compared to the control sample, and hydrolysis of ester bonds. These preliminary results are encouraging for further investigation on the ability of the microbiome associated with plankton to biodegrade polyesters, such as PBSA, and increasing knowledge on microorganisms involved in bioplastic degradation in marine environment.
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Biodegradação Ambiental , Copépodes , Microbiota , Animais , Bactérias/metabolismo , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Poliésteres/metabolismo , Adipatos/metabolismo , Polímeros/metabolismo , RNA Ribossômico 16S/genética , Plâncton/metabolismo , Butileno GlicóisRESUMO
This study covers the preparation of microspheres for the controlled and targeted release of paclitaxel, using novel degradable polymers as carrier materials. Paclitaxel-loaded microspheres were prepared by oil-in-water single-emulsion solvent extraction/evaporation technique by using a series of polyurethanes and a block copolymer; the physicochemical properties of these polymers were modulated by changing nature and composition of their structural units. The obtained microparticles showed a regular morphology and properties (diameter: 1-100 µm; resuspension index: 18.8-100%; encapsulation efficiency: 26.6-97.2%) depending on polymer hydrophilicity and emulsifier used. In vitro release curves showed in all cases almost zero-order kinetics after an initial low burst effect (from 1 to 8.4%), which is required to minimize the drug side effects. This work also proposes a novel strategy to combine a controlled and a targeted release through the functionalization of the polymer matrix with peptide sequences. An RGD-functionalized polyurethane was used to successfully prepare paclitaxel-loaded microparticles. Studies on the preparation of polymer microspheres are reported.
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Portadores de Fármacos/química , Microesferas , Oligopeptídeos/química , Paclitaxel/química , Poliuretanos/química , Preparações de Ação Retardada , Interações Hidrofóbicas e Hidrofílicas , Tamanho da PartículaRESUMO
Type-1 diabetes is one of the most prevalent metabolic disorders worldwide. It results in a significant lack of insulin production by the pancreas and the ensuing hyperglycemia, which needs to be regulated through a tailored administration of insulin throughout the day. Recent studies have shown great advancements in developing an implantable artificial pancreas. However, some improvements are still required, including the optimal biomaterials and technologies to produce the implantable insulin reservoir. Here, we discuss the employment of two types of cyclic olefin copolymers (Topas 5013L-10 and Topas 8007S-04) for an insulin reservoir fabrication. After a preliminary thermomechanical analysis, Topas 8007S-04 was selected as the best material to fabricate a 3D-printed insulin reservoir due to its higher strength and lower glass transition temperature (Tg). Fiber deposition modeling was used to manufacture a reservoir-like structure, which was employed to assess the ability of the material to prevent insulin aggregation. Although the surface texture presents a localized roughness, the ultraviolet analysis did not detect any significant insulin aggregation over a timeframe of 14 days. These interesting results make Topas 8007S-04 cyclic olefin copolymer a potential candidate biomaterial for fabricating structural components in an implantable artificial pancreas.
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Over the past decade, a large number of strategies and technologies have been developed to reduce heart failure progression. Among these, cardiac tissue engineering is one of the most promising. Aim of this study is to develop a 3D scaffold to treat cardiac failure. A new three-block copolymer, obtained from δ-valerolactone and polyoxyethylene, was synthesised under high vacuum without catalyst. Copolymer/gelatine blends were microfabricated to obtain a ECM-like geometry. Structures were studied under morphological, mechanical, degradation and biological aspects. To prevent left ventricular remodelling, constructs were biofunctionalises with molecularly imprinted nanoparticles towards the matrix metalloproteinase MMP-9. Results showed that materials are able to reproduce the ECM structure with high resolution, mechanical properties were in the order of MPa similar to those of the native myocardium and cell viability was verified. Nanoparticles showed the capability to rebind MMP-9 (specific rebinding 18.67) and to be permanently immobilised on the scaffold surface.
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Biomimética , Infarto do Miocárdio/patologia , Polímeros/química , Alicerces Teciduais , Remodelação Ventricular , Humanos , Metaloproteinase 9 da Matriz/metabolismo , Impressão Molecular , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
The aim of this work was the morphological, physicochemical, mechanical and biological characterization of a new composite system, based on gelatin, gellan and hydroxyapatite, and mimicking the composition of natural bone. Porous scaffolds were prepared by freeze-drying technique, under three different conditions of freezing. The morphological analysis showed a homogeneous porosity, with well interconnected pores, for the sample which underwent a more rapid freezing. The elastic modulus of the same sample was close to that of the natural bone. The presence of interactions among the components was demonstrated through the physicochemical investigation. In addition, the infrared chemical imaging analysis pointed out the similarity among the composite scaffold and the natural bone, in terms of chemical composition, homogeneity, molecular interactions and structural conformation. Preliminary biological characterization showed a good adhesion and proliferation of human mesenchymal stem cells.
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Desenvolvimento Ósseo , Substitutos Ósseos , Durapatita/química , Gelatina/química , Nanocompostos , Polissacarídeos Bacterianos/química , Varredura Diferencial de Calorimetria , Sequência de Carboidratos , Liofilização , Humanos , Microscopia Eletrônica de Varredura , Dados de Sequência Molecular , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
The present work aimed at the production and characterization of small caliber biomimetic and bioactive tubular scaffolds, which are able to favor the endothelialization process, and therefore potentially be suitable for vascular tissue engineering. The tubular scaffolds were produced using a specially designed mold, starting from a gelatin/gellan/elastin (GGE) blend, selected to mimic the composition of the extracellular matrix of native blood vessels. GGE scaffolds were obtained through freeze-drying and subsequent cross-linking. To obtain systems capable of promoting endothelization, the scaffolds were functionalized using two different bioactive peptides, Gly-Arg-Gly-Asp-Ser-Pro (GRGSDP) and Arg-Glu-Asp-Val (REDV). A complete physicochemical, mechanical, functional, and biological characterization of the developed scaffolds was performed. GGE scaffolds showed a good porosity, which could promote cell infiltration and proliferation and a dense external surface, which could avoid bleeding. Moreover, developed scaffolds showed good hydrophilicity, an elastic behavior similar to natural vessels, suitability for sterilization by an ISO accepted treatment, and an adequate suture retention strength. In vitro cell culture tests showed no cytotoxic activity against 3T3 fibroblasts. The functionalization with the REDV peptide favored the adhesion and growth of endothelial cells, while GRGDSP-modified scaffolds represented a better substrate for fibroblasts.
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The recent advances in nanotechnology are revolutionizing preventive and therapeutic approaches to treating cardiovascular diseases. Controlling the extracellular matrix metalloproteinase (MMP) activation and expression in the failing human left ventricular myocardium represents a significant therapeutic target for heart disease. In this study, we used molecularly imprinting polymers (MIPs) to restore the correct balance between MMPs and their tissue inhibitors (TIMPs), and explored the potential of this technique exhaustively through chemical synthesis, physicochemical and biological characterizations, and computational chemistry methods. By molecular dynamics simulations based on classical force fields, we simulated the early stages of the imprinting process in solution disclosing the pivotal interaction established between the monomers and the MMP9 protein template. The average interaction energies of methacrylic acid (MAA) and poly (ethylene glycol) ethyl ether methacrylate (PEG) units were in the ranges 17-22 and 30-37 kcal/mol, respectively. At low coverage, the PEG monomers seemed firmly anchored to the protein surface and were not displaced by water, while only about 20% of MAA was replaced by water. The synthesis of MIPs was successfully with a monomer conversion higher than 99% and the production of spherical particles with average diameter of 344 ± 33 nm. HPLC analysis showed a specific recognition factor of MMP9 on MIPs of about 1.3. FT-IR Chemical Imaging confirmed the mechanisms necessary to generate a "selective memory" of the MIPs towards the enzyme. HPLC results indicated that the rebound amount of both TIMP1 and MMP2 to MIPs is lower than that of the template, showing a selectivity factor of 2.1 and 2.3, respectively. Preliminary tests on the effect of MIPs on H9C2 cells revealed that this treatment has no cytotoxic effects.
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The interactions of Type I acid soluble collagen (Col) with both carbonate-free hydroxyapatite (HA(1100)) and carbonate-rich one (CHA) were investigated. The aim was to ascertain whether the increase of bone CO(3) (2-) with ageing could relate to the disease known as osteoporosis. HA(1100)-Col and CHA-Col composites with various ratios were prepared and examined. Scanning electron microscopy and differential scanning calorimetry showed a stronger adhesion of the Col matrix to the granules of HA(1100) than to those of CHA. FT-IR spectroscopy showed that with HA(1100) both multiple hydrogen bonds of Col peptide -NH groups with HA PO(4) (3-), and electrochemical interactions between Col peptide -C=O groups and HA Ca(2+) were present. In the presence of CO(3) (2-), the interactions between -NH and phosphate were diminished, and Ca(2+) interacted more strongly with CO(3) (2-) than with peptide -C=O, so causing a separation between the two components of the bone extra-cellular matrix. The results obtained strengthen the hypothesis that the substitution of PO(4) (3-) ions by CO(3) (2-) ions in the HA lattice might be a significant component of osteoporosis, although further investigation is needed.
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Envelhecimento , Osso e Ossos/patologia , Durapatita/química , Osteoporose/metabolismo , Varredura Diferencial de Calorimetria/métodos , Físico-Química/métodos , Matriz Extracelular/patologia , Humanos , Ligação de Hidrogênio , Íons , Microscopia Eletrônica de Varredura/métodos , Modelos Químicos , Espectrofotometria Infravermelho/métodos , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Estresse Mecânico , TemperaturaRESUMO
Chemotherapeutics represent the standard treatment for a wide range of cancers. However, these agents also affect healthy cells, thus leading to severe off-target effects. Given the non-selectivity of the commonly used drugs, any increase in the selective tumor tissue uptake would represent a significant improvement in cancer therapy. Recently, the use of gene therapy to completely remove the lesion and avoid the toxicity of chemotherapeutics has become a tendency in oncotherapy. Ideally, the genetic material must be safely transferred from the site of administration to the target cells, without involving healthy tissues. This can be achieved by encapsulating genes into non-viral carriers and modifying their surface with ligands with high selectivity and affinity for a relevant receptor on the target cells. Hence, in this work we evaluate the use of terpolymer-based nanocapsules for the targeted delivery of DNA toward cancer cells. The surface of the nanocapsules is decorated with folic acid to actively target the folate receptors overexpressed on a variety of cancer cells. The nanocapsules demonstrate a good ability of encapsulating and releasing DNA. Moreover, the presence of the targeting moieties on the surface of the nanocapsules favors cell uptake, opening up the possibility of more effective therapies.
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The use of injectable scaffolds to repair the infarcted heart is receiving great interest. Thermosensitive polymers, in situ polymerization, in situ cross-linking, and self-assembling peptides are the most investigated approaches to obtain injectability.Aim of the present work was the preparation and characterization of a novel bioactive scaffold, in form of injectable microspheres, for cardiac repair. Gellan/gelatin microspheres were prepared by a water-in-oil emulsion and loaded by adsorption with Insulin-like growth factor 1 to promote tissue regeneration. Obtained microspheres underwent morphological, physicochemical and biological characterization, including cell culture tests in static and dynamic conditions and in vivo tests. Morphological analysis of the microspheres showed a spherical shape, a microporous surface and an average diameter of 66 ± 17µm (under dry conditions) and 123 ± 24 µm (under wet conditions). Chemical Imaging analysis pointed out a homogeneous distribution of gellan, gelatin and Insulin-like growth factor-1 within the microsphere matrix. In vitro cell culture tests showed that the microspheres promoted rat cardiac progenitor cells adhesion, and cluster formation. After dynamic suspension culture within an impeller-free bioreactor, cells still adhered to microspheres, spreading their cytoplasm over microsphere surface. Intramyocardial administration of microspheres in a cryoinjury rat model attenuated chamber dilatation, myocardial damage and fibrosis and improved cell homing.Overall, the findings of this study confirm that the produced microspheres display morphological, physicochemical, functional and biological properties potentially adequate for future applications as injectable scaffold for cardiac tissue engineering.
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Coração/efeitos dos fármacos , Fator de Crescimento Insulin-Like I/administração & dosagem , Microesferas , Miocárdio/patologia , Alicerces Teciduais , Animais , Materiais Biocompatíveis , Reatores Biológicos , Adesão Celular , Meios de Cultura , Injeções , Insulina/metabolismo , Cinética , Masculino , Microfluídica , Microscopia Eletrônica de Varredura , Infarto do Miocárdio/terapia , Polímeros/química , Ratos , Ratos Wistar , Regeneração , Células-Tronco/citologia , Engenharia Tecidual/métodosRESUMO
In recent years, there has been an increasing interest toward the covalent binding of bioactive peptides from extracellular matrix proteins on scaffolds as a promising functionalization strategy in the development of biomimetic matrices for tissue engineering. A totally new approach for scaffold functionalization with peptides is based on Molecular Imprinting technology. In this work, imprinted particles with recognition properties toward laminin and fibronectin bioactive moieties were synthetized and used for the functionalization of biomimetic sponges, which were based on a blend of alginate, gelatin, and elastin. Functionalized sponges underwent a complete morphological, physicochemical, mechanical, functional, and biological characterization. Micrographs of functionalized sponges showed a highly porous structure and a quite homogeneous distribution of imprinted particles on their surface. Infrared and thermal analyses pointed out the presence of interactions between blend components. Biodegradation and mechanical properties appeared adequate for the aimed application. The results of recognition tests showed that the deposition on sponges did not alter the specific recognition and binding behavior of imprinted particles. In vitro biological characterization with cardiac progenitor cells showed that early cell adherence was promoted. In vivo analysis showed that developed scaffolds improved cardiac progenitor cell adhesion and differentiation toward myocardial phenotypes.
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Cystic fibrosis (CF) is a progressive genetic disease caused by mutations in the gene that produces the CF transmembrane conductance regulator (CFTR) protein. The malfunction of the CFTR protein causes a thick buildup of mucus in the lungs that clogs the airways and traps bacteria, thus leading to infections, extensive lung damage and respiratory failure. Micro-delivery systems are currently being investigated as an efficient way to cross the viscous and complex architecture of the CF mucus. In this study, we produced synthetic and natural microparticles (MPs) based on poly(dllactidecoglycolide) (PLGA) or gellan gum through tailored water/oil emulsion procedures. Morphological and physico-chemical characterizations were carried out on both classes of MPs showing particles having diameters within suitable ranges to reach the CF airways. In vitro biocompatibility tests were also performed on both MPs using a human lung cancer cell line (A549) demonstrating that treatment with MPs induces no cytotoxic effects. Both classes of MPs were loaded with a mucolytic agent (Nacetyl cysteine, NAC) and their release kinetics evaluated using high performance liquid chromatography (HPLC). The analysis pointed out that the amount of NAC released from MPs resulted in a dose-dependent increment, with a rapid release kinetic to satisfy the requirement for inducing an early mucus degradation. Finally, mucolytic action of NAC-loaded MPs was evaluated in an artificial sputum model through its rheological analysis obtaining the lowest viscosity profile after the addition of drug-loaded MPs. Taken together, gained results allowed us to select suitable MPs as potential drug targeting platforms having a mucolytic action for CF treatment.
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Materiais Biocompatíveis/metabolismo , Fibrose Cística/metabolismo , Muco/metabolismo , Células A549 , Adulto , Proliferação de Células , Cromatografia Líquida de Alta Pressão , Sistemas de Liberação de Medicamentos/métodos , Humanos , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/metabolismo , Escarro/metabolismoRESUMO
The protection from ischaemia-reperfusion-associated myocardial infarction worsening remains a big challenge. We produced a bioartificial 3D cardiac patch with cardioinductive properties on stem cells. Its multilayer structure was functionalised with clinically relevant doses of adenosine. We report here the first study on the potential of these cardiac patches in the controlled delivery of adenosine into the in vivo ischaemic-reperfused pig heart. A Fourier transform infrared chemical imaging approach allowed us to perform a characterisation, complementary to the histological and biochemical analyses on myocardial samples after in vivo patch implantation, increasing the number of investigations and results on the restricted number of pigs (n = 4) employed in this feasibility step. In vitro tests suggested that adenosine was completely released by a functionalised patch, a data that was confirmed in vivo after 24 hr from patch implantation. Moreover, the adenosine-loaded patch enabled a targeted delivery of the drug to the ischaemic-reperfused area of the heart, as highlighted by the activation of the pro-survival signalling reperfusion injury salvage kinases pathway. At 3 months, though limited to one animal, the used methods provided a picture of a tissue in dynamic conditions, associated to the biosynthesis of new collagen and to a non-fibrotic outcome of the healing process underway. The synergistic effect between the functionalised 3D cardiac patch and adenosine cardioprotection might represent a promising innovation in the treatment of reperfusion injury. As this is a feasibility study, the clinical implications of our findings will require further in vivo investigation on larger numbers of ischaemic-reperfused pig hearts.
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Adenosina , Gelatina , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Miocárdio , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Adenosina/química , Adenosina/farmacologia , Animais , Modelos Animais de Doenças , Implantes de Medicamento/química , Implantes de Medicamento/farmacologia , Feminino , Gelatina/química , Gelatina/farmacologia , Traumatismo por Reperfusão Miocárdica/metabolismo , Traumatismo por Reperfusão Miocárdica/patologia , Miocárdio/metabolismo , Miocárdio/patologia , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/farmacologia , SuínosRESUMO
In the present work, a new combination of synthetic and natural biomaterials is proposed for bone tissue engineering (BTE) applications. In order to mimic the inorganic and organic phases of bone extracellular matrix (ECM), a bioactive glass-ceramic deriving from a SiO2-P2O5-CaO-MgO-Na2O-K2O parent glass, acting as a substrate in form of a slice, was surface-functionalised with a type I collagen-based coating. In particular, the collagen was blended with a water soluble polyurethane (PUR), synthesised from poly(ethylene glycol), 1,6-hexamethylene diisocyanate and N-BOC-serinol. The PUR was designed to expose amino groups on the polymeric chain, which can be exploited for the blend stabilisation through crosslinking. The newly synthesised PUR demonstrated to be non-cytotoxic, as assessed by a biological test with MG-63 osteoblast-like cells. The collagen/PUR blend showed good biocompatibility as well. The polymeric coating on the glass-ceramic samples was produced by surface-silanisation, followed by further chemical grafting of the blend, using genipin as a crosslinker. The glass-ceramic surface was characterised at each functionalisation step, demonstrating that the procedure allowed obtaining a covalent link between the blend and the substrate. Finally, biological tests performed using human periosteal derived precursor cells demonstrated that the proposed polymer-coated material was a good substrate for bone cell adhesion and growth, and a good candidate to mimic the composite nature of the bone ECM.
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Osso e Ossos/metabolismo , Cerâmica/química , Materiais Revestidos Biocompatíveis/química , Colágeno/química , Osteoblastos/metabolismo , Poliuretanos/química , Engenharia Tecidual , Osso e Ossos/citologia , Linhagem Celular Tumoral , Humanos , Osteoblastos/citologiaRESUMO
Tissue engineering has emerged as a viable approach to treat disease or repair damage in tissues and organs. One of the key elements for the success of tissue engineering is the use of a scaffold serving as artificial extracellular matrix (ECM). The ECM hosts the cells and improves their survival, proliferation, and differentiation, enabling the formation of new tissue. Here, we propose the development of a class of protein/polysaccharide-based porous scaffolds for use as ECM substitutes in cardiac tissue engineering. Scaffolds based on blends of a protein component, collagen or gelatin, with a polysaccharide component, alginate, were produced by freeze-drying and subsequent ionic and chemical crosslinking. Their morphological, physicochemical, and mechanical properties were determined and compared with those of natural porcine myocardium. We demonstrated that our scaffolds possessed highly porous and interconnected structures, and the chemical homogeneity of the natural ECM was well reproduced in both types of scaffolds. Furthermore, the alginate/gelatin (AG) scaffolds better mimicked the native tissue in terms of interactions between components and protein secondary structure, and in terms of swelling behavior. The AG scaffolds also showed superior mechanical properties for the desired application and supported better adhesion, growth, and differentiation of myoblasts under static conditions. The AG scaffolds were subsequently used for culturing neonatal rat cardiomyocytes, where high viability of the resulting cardiac constructs was observed under dynamic flow culture in a microfluidic bioreactor. We therefore propose our protein/polysaccharide scaffolds as a viable ECM substitute for applications in cardiac tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 769-781, 2018.
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Materiais Biomiméticos/química , Matriz Extracelular/metabolismo , Coração/fisiologia , Polissacarídeos/química , Proteínas/química , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Animais , Reatores Biológicos , Bovinos , Linhagem Celular , Proliferação de Células , Forma Celular , Módulo de Elasticidade , Hidrólise , Cinética , Microfluídica , Mioblastos/citologia , Ratos , Espectroscopia de Infravermelho com Transformada de Fourier , SuínosRESUMO
INTRODUCTION: Injectable scaffolds are emerging as a promising strategy in the field of myocardial tissue engineering. Among injectable scaffolds, microparticles have been poorly investigated. The goal of this study was the development of novel gelatin/gellan microparticles that could be used as an injectable scaffold to repair the infarcted myocardium. In particular, the effect of particle size on cardiac progenitor cell response was investigated. METHODS: Particles were produced by a water-in-oil emulsion method. Phosphatidylcholine was used as a surfactant. Particles with different diameter ranges (125-300 µm and 350-450 µm) were fabricated using two different surfactant concentrations. Morphological, physicochemical, and functional characterizations were carried out. Cardiac progenitor cell adhesion and growth on microparticles were tested both in static and dynamic suspension culture conditions. RESULTS: Morphological analysis of the produced particles showed a spherical shape and porous surface. The hydrophilicity of particle matrix and the presence of intermolecular interactions between gellan and gelatin were pointed out by the physicochemical characterization. A weight loss of 75 ± 5 % after 90 days of hydrolytic degradation was observed. Injectability through a narrow needle (26 G) and persistence of the microparticles at the injection site were preliminarily verified by ex vivo test. In vitro cell culture tests showed a preservation of rat cardiac progenitor biologic properties and indicated a preferential cell adherence to microparticles with a smaller size. CONCLUSION: Overall, the obtained results indicate that the produced gelatin/gellan microparticles could be potentially employed as injectable scaffolds for myocardial regeneration.