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Cardiovascular diseases remain a leading cause of mortality globally, with atherosclerosis representing a significant pathological means, often leading to myocardial infarction. Coronary artery bypass surgery, a common procedure used to treat coronary artery disease, presents challenges due to the limited autologous tissue availability or the shortcomings of synthetic grafts. Consequently, there is a growing interest in tissue engineering approaches to develop vascular substitutes. This review offers an updated picture of the state of the art in vascular tissue engineering, emphasising the design of scaffolds and dynamic culture conditions following a biomimetic approach. By emulating native vessel properties and, in particular, by mimicking the three-layer structure of the vascular wall, tissue-engineered grafts can improve long-term patency and clinical outcomes. Furthermore, ongoing research focuses on enhancing biomimicry through innovative scaffold materials, surface functionalisation strategies, and the use of bioreactors mimicking the physiological microenvironment. Through a multidisciplinary lens, this review provides insight into the latest advancements and future directions of vascular tissue engineering, with particular reference to employing biomimicry to create systems capable of reproducing the structure-function relationships present in the arterial wall. Despite the existence of a gap between benchtop innovation and clinical translation, it appears that the biomimetic technologies developed to date demonstrate promising results in preventing vascular occlusion due to blood clotting under laboratory conditions and in preclinical studies. Therefore, a multifaceted biomimetic approach could represent a winning strategy to ensure the translation of vascular tissue engineering into clinical practice.
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Background: Abdominal hernia repair surgeries involve the fixation of a surgical mesh to the abdominal wall with different means such as suture, tacks, and glues. Currently, the most effective mesh fixation system is still debated. This review compares outcomes of mesh fixation in different surgical procedures, aiding surgeons in identifying the optimal technique. Methods: A meta-analysis was conducted according to PRISMA guidelines. Articles published between January 2003 and January 2023 were searched in electronic databases. Randomized controlled trials (RCTs) comparing mesh fixation with cyanoacrylate-based or fibrin glues with classical fixation techniques (sutures, tacks) in open and laparoscopic procedures were included. Results: 17 RCTs were identified; the cumulative study population included 3919 patients and a total of 3976 inguinal hernias. Cyanoacrylate-based and fibrin glues were used in 1639 different defects, suture and tacks in 1912 defects, self-gripping mesh in 404 cases, and no mesh fixation in 21 defects. Glue fixation resulted in lower early postoperative pain, and chronic pain occurred less frequently. The incidence of hematoma was lower with glue fixation than with mechanical fixation. Recurrence rate, seroma formation, operative and hospitalization time showed no significant differences; but significantly, a higher number of people in the glue group returned to work by 15- and 30-days after surgery when compared to the tacker and suture groups in the same time frame. Conclusion: Cyanoacrylate and fibrin glue may be effective in reducing early and chronic pain and hematoma incidence without increasing the recurrence rate, the seroma formation, or the operative and hospitalization time.
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A multitude of sight-threatening retinal diseases, affecting hundreds of millions around the globe, lack effective pharmacological treatments due to ocular barriers and common drug delivery limitations. Polymeric nanoparticles (PNPs) are versatile drug carriers with sustained drug release profiles and tunable physicochemical properties which have been explored for ocular drug delivery to both anterior and posterior ocular tissues. PNPs can incorporate a wide range of drugs and overcome the challenges of conventional retinal drug delivery. Moreover, PNPs can be engineered to respond to specific stimuli such as ultraviolet, visible, or near-infrared light, and allow precise spatiotemporal control of the drug release, enabling tailored treatment regimens and reducing the number of required administrations. The objective of this study is to emphasize the therapeutic potential of light-triggered drug-loaded polymeric nanoparticles to treat retinal diseases through an exploration of ocular pathologies, challenges in drug delivery, current production methodologies and recent applications. Despite challenges, light-responsive PNPs hold the promise of substantially enhancing the treatment landscape for ocular diseases, aiming for an improved quality of life for patients.
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Myocardial infarction is one of the major causes of mortality as well as morbidity around the world. Currently available treatment options face a number of drawbacks, hence cardiac tissue engineering, which aims to bioengineer functional cardiac tissue, for application in tissue repair, patient specific drug screening and disease modeling, is being explored as a viable alternative. To achieve this, an appropriate combination of cells, biomimetic scaffolds mimicking the structure and function of the native tissue, and signals, is necessary. Among scaffold fabrication techniques, three-dimensional printing, which is an additive manufacturing technique that enables to translate computer-aided designs into 3D objects, has emerged as a promising technique to develop cardiac patches with a highly defined architecture. As a further step toward the replication of complex tissues, such as cardiac tissue, more recently 3D bioprinting has emerged as a cutting-edge technology to print not only biomaterials, but also multiple cell types simultaneously. In terms of bioinks, biomaterials isolated from natural sources are advantageous, as they can provide exceptional biocompatibility and bioactivity, thus promoting desired cell responses. An ideal biomimetic cardiac patch should incorporate additional functional properties, which can be achieved by means of appropriate functionalization strategies. These are essential to replicate the native tissue, such as the release of biochemical signals, immunomodulatory properties, conductivity, enhanced vascularization and shape memory effects. The aim of the review is to present an overview of the current state of the art regarding the development of biomimetic 3D printed natural biomaterial-based cardiac patches, describing the 3D printing fabrication methods, the natural-biomaterial based bioinks, the functionalization strategies, as well as the in vitro and in vivo applications.
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BACKGROUND: Agrifood waste products are often considered rich sources of bioactive compounds that can be conveniently recovered. Due to these peculiar characteristics, the study of these waste products is attracting great interest in nutraceutical research. Olive mill wastewaters (OMWWs) are generated by extra virgin olive oil (EVOO) production, and they pose environmental challenges due to their disposal. This study aimed to characterize the polyphenolic profile and to evaluate the nutraceutical properties of OMWW extracts from two Tuscan olive cultivars, Leccino (CL) and Frantoio (CF), collected during different time points in EVOO production. METHOD: After a liquid-liquid extraction, the HPLC and LC-MS/MS analysis of OMWW extracts confirmed the presence of 18 polyphenolic compounds. RESULTS: The polyphenol composition varied between the cultivars and during maturation stages. Notably, oleacein was detected at remarkably high levels in CL1 and CF1 extracts (314.628 ± 19.535 and 227.273 ± 3.974 µg/mg, respectively). All samples demonstrated scavenging effects on free radicals (DPPH and ABTS assays) and an anti-inflammatory potential by inhibiting cyclooxygenase (COX) enzymes. CONCLUSIONS: This study highlights the nutraceutical potential of OMWW extracts, emphasizing their antioxidant, antiradical, and anti-inflammatory activities. The results demonstrate the influence of olive cultivar, maturation stage, and extraction process on the polyphenolic composition and the bioactivity of OMWW extracts. These findings support a more profitable reuse of OMWW as an innovative, renewable, and low-cost source of dietary polyphenols with potential applications as functional ingredients in the development of dietary supplements, as well as in the pharmaceutical and cosmetics industries.
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Olea , Águas Residuárias , Polifenóis , Cromatografia Líquida , Espectrometria de Massas em Tandem , Suplementos Nutricionais , Resíduos , Extratos Vegetais/farmacologiaRESUMO
Natural polymers, thanks to their intrinsic biocompatibility and biomimicry, have been largely investigated as scaffold materials for tissue engineering applications. Traditional scaffold fabrication methods present several limitations, such as the use of organic solvents, the obtainment of a non-homogeneous structure, the variability in pore size and the lack of pore interconnectivity. These drawbacks can be overcome using innovative and more advanced production techniques based on the use of microfluidic platforms. Droplet microfluidics and microfluidic spinning techniques have recently found applications in the field of tissue engineering to produce microparticles and microfibers that can be used as scaffolds or as building blocks for three-dimensional structures. Compared to standard fabrication technologies, microfluidics-based ones offer several advantages, such as the possibility of obtaining particles and fibers with uniform dimensions. Thus, scaffolds with extremely precise geometry, pore distribution, pore interconnectivity and a uniform pores size can be obtained. Microfluidics can also represent a cheaper manufacturing technique. In this review, the microfluidic fabrication of microparticles, microfibers and three-dimensional scaffolds based on natural polymers will be illustrated. An overview of their applications in different tissue engineering fields will also be provided.
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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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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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The purpose of this work is to highlight the effects of ionizing radiation on the genetic material in higher plants by assessing both adaptive processes as well as the evolution of plant species. The effects that the ionizing radiation has on greenery following a nuclear accident, was examined by taking the Chernobyl Nuclear Power Plant disaster as a case study. The genetic and evolutionary effects that ionizing radiation had on plants after the Chernobyl accident were highlighted. The response of biota to Chernobyl irradiation was a complex interaction among radiation dose, dose rate, temporal and spatial variation, varying radiation sensitivities of the different plants' species, and indirect effects from other events. Ionizing radiation causes water radiolysis, generating highly reactive oxygen species (ROS). ROS induce the rapid activation of detoxifying enzymes. DeoxyriboNucleic Acid (DNA) is the object of an attack by both, the hydroxyl ions and the radiation itself, thus triggering a mechanism both direct and indirect. The effects on DNA are harmful to the organism and the long-term development of the species. Dose-dependent aberrations in chromosomes are often observed after irradiation. Although multiple DNA repair mechanisms exist, double-strand breaks (DSBs or DNA-DSBs) are often subject to errors. Plants DSBs repair mechanisms mainly involve homologous and non-homologous dependent systems, the latter especially causing a loss of genetic information. Repeated ionizing radiation (acute or chronic) ensures that plants adapt, demonstrating radioresistance. An adaptive response has been suggested for this phenomenon. As a result, ionizing radiation influences the genetic structure, especially during chronic irradiation, reducing genetic variability. This reduction may be associated with the fact that particular plant species are more subject to chronic stress, confirming the adaptive theory. Therefore, the genomic effects of ionizing radiation demonstrate their likely involvement in the evolution of plant species.
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Adaptação Fisiológica , Acidente Nuclear de Chernobyl , Desastres , Fenômenos Fisiológicos Vegetais , Monitoramento de Radiação , Radioatividade , Plantas , Radiação IonizanteAssuntos
Alginatos/química , Avidina/química , Materiais Biomiméticos/química , Biotina/química , Gelatina/química , Poríferos/química , Alicerces Teciduais/química , Animais , Materiais Biomiméticos/metabolismo , Adesão Celular , Proliferação de Células , Elasticidade , Humanos , Interações Hidrofóbicas e Hidrofílicas , Masculino , Miocárdio/citologia , Polidioxanona/química , Porosidade , Implantação de Prótese , Ratos , Somatomedinas/química , Somatomedinas/metabolismo , Células-Tronco , Propriedades de Superfície , Engenharia Tecidual , ViscosidadeRESUMO
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
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.
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Microesferas , Miocárdio/citologia , Miócitos Cardíacos/citologia , Células-Tronco/citologia , Animais , Materiais Biocompatíveis , Adesão Celular , Proliferação de Células , Células Cultivadas , Emulsões , Gelatina/química , Miócitos Cardíacos/fisiologia , Tamanho da Partícula , Polissacarídeos Bacterianos/química , Porosidade , Ratos , Células-Tronco/fisiologia , Engenharia Tecidual/métodos , Alicerces TeciduaisRESUMO
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
The development of biomaterials for cardiac tissue engineering (CTE) is challenging, primarily owing to the requirement of achieving a surface with favourable characteristics that enhances cell attachment and maturation. The biomaterial surface plays a crucial role as it forms the interface between the scaffold (or cardiac patch) and the cells. In the field of CTE, synthetic polymers (polyglycerol sebacate, polyethylene glycol, polyglycolic acid, poly-l-lactide, polyvinyl alcohol, polycaprolactone, polyurethanes and poly(N-isopropylacrylamide)) have been proven to exhibit suitable biodegradable and mechanical properties. Despite the fact that they show the required biocompatible behaviour, most synthetic polymers exhibit poor cell attachment capability. These synthetic polymers are mostly hydrophobic and lack cell recognition sites, limiting their application. Therefore, biofunctionalization of these biomaterials to enhance cell attachment and cell material interaction is being widely investigated. There are numerous approaches for functionalizing a material, which can be classified as mechanical, physical, chemical and biological. In this review, recent studies reported in the literature to functionalize scaffolds in the context of CTE, are discussed. Surface, morphological, chemical and biological modifications are introduced and the results of novel promising strategies and techniques are discussed.
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Materiais Biocompatíveis/química , Miocárdio , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Animais , Adesão Celular , HumanosRESUMO
Tissue engineering has the potential to supply constructs capable of restoring the normal function of native tissue following injury. Poly(L-lactic acid) (PLLA) scaffolds are amongst the most commonly used biodegradable polymers in tissue engineering and previous studies performed on ovine fibroblasts have showed that addition of gelatin creates a favorable hydrophilic microenvironment for the growth of these cells. The attractiveness of using mesenchymal stromal cells (MSCs) in tissue regeneration is that they are able to differentiate into several lines including osteoblasts. In this study, we investigated the ability of gelatin/PLLA sponges to support the adhesion, proliferation, and osteogenic differentiation of human MSCs isolated from the bone marrow of four donors. [Figure: see text].
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Gelatina , Ácido Láctico , Células-Tronco Mesenquimais/citologia , Osteogênese , Polímeros , Engenharia Tecidual/métodos , Diferenciação Celular , Proliferação de Células , Humanos , Poliésteres , Células Estromais/citologiaRESUMO
In this paper a device, based on urease-loaded microspheres, is presented. The first task of this work was the optimization of a procedure for the alginate microspheres realization, having a radius as close as possible to the optimal one necessary to achieve the maximum enzyme exploitation. This optimal radius was calculated theoretically through a mathematical model which describes the concentration of substrate (urea) inside the microspheres on the assumption of a diffusion-reaction mechanism. The enzyme-loaded microspheres were successfully tested in a prototypal device aimed at the depletion of urea from a circulating fluid simulating blood flow: the results showed that urea concentration in the circulating fluid drops down to less than 25% of the initial value after 5 h.
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Alginatos/química , Portadores de Fármacos/química , Microesferas , Ureia/metabolismo , Urease/uso terapêutico , Alginatos/uso terapêutico , Enzimas Imobilizadas/administração & dosagem , Enzimas Imobilizadas/metabolismo , Enzimas Imobilizadas/uso terapêutico , Ácido Glucurônico/química , Ácido Glucurônico/uso terapêutico , Ácidos Hexurônicos/química , Ácidos Hexurônicos/uso terapêutico , Humanos , Nefropatias/terapia , Tamanho da Partícula , Ureia/sangue , Urease/administração & dosagem , Urease/metabolismoRESUMO
Tissue engineering scaffolds should be able to reproduce optimal microenvironments in order to support cell attachment, three-dimensional growth, migration and, regarding fibroblasts, must also promote extracellular matrix production. Various bioactive molecules are employed in the preparation of spongy scaffolds to obtain biomimetic matrices by either surface-coating or introducing them into the bulk composition of the biomaterial. The biomimetic properties of a spongy matrix composed of PVA combined with the natural component gelatine were evaluated by culturing human gingival fibroblasts on the scaffold. Cell adhesion, morphology and distribution within the scaffold were assessed by histology and electron microscopy; viability and metabolic activity as well as extracellular matrix production were analyzed by MTT assay, cytochemistry and immunocytochemistry. Fibroblasts interacted positively with PVA/gelatine. They adhered to the PVA/gelatine matrix in which they had good spreading activity and active metabolism; fibroblasts were also able to produce extracellular matrix molecules (type I collagen, fibronectin and laminin) compared to bi-dimensionally grown cells. The in situ creation of a biological matrix by human fibroblasts together with the ability to produce growth factor TGF-beta1 and the intracellular signal transduction molecule RhoA, suggests that this kind of PVA/gelatine sponge may represent a suitable support for in vitro extracellular matrix production and connective tissue regeneration.
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Matriz Extracelular/metabolismo , Fibroblastos/fisiologia , Gelatina , Gengiva/citologia , Álcool de Polivinil , Engenharia Tecidual/métodos , Adesão Celular , Células Cultivadas , Fibroblastos/citologia , Fibroblastos/ultraestrutura , Humanos , Imuno-Histoquímica , Microscopia Eletrônica/métodos , Fator de Crescimento Transformador beta1/metabolismo , Proteína rhoA de Ligação ao GTP/metabolismoRESUMO
Biodegradable synthetic polymers such as poly(lactic acid) (PLA) are widely used to prepare scaffolds for cell transplantation and tissue growth, using different techniques set up for the purpose. However the poor hydrophilicity of these polymers represents the main limitation to their use as scaffolds because it causes a low affinity for the cells. An effective way to solve this problem could be represented by the addition of biopolymers that are in general highly hydrophilic. The present work concerns porous biodegradable sponge-like systems based on poly(L-lactic acid) (PLLA) and gelatine. Morphology and porosity characteristics of the sponges were studied by scanning electron microscopy and mercury intrusion porosimetry respectively. Blood compatibility was investigated by bovine plasma fibrinogen (BPF) adsorption test and platelet adhesion test (PAT). The cell culture method was used in order to evaluate the ability of the matrices to work as scaffolds for tissue regeneration. The obtained results indicate that the sponges have interesting porous characteristics, good blood compatibility and above all good ability to support cell adhesion and growth. In fact viable and metabolically active animal cells were found inside the sponges after 8 weeks in culture. On this basis the systems produced seem to be good candidates as scaffolds for tissue regeneration.