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Andrea E Sprio,
Federica Di Scipio,
Stefania Raimondo,
Paolina Salamone, Francesca Pagliari,
Stefania Pagliari,
Anna Folino,
Giancarlo Forte,
Stefano Geuna,
Paolo Di Nardo,
Giovanni N Berta
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ABSTRACT: The stemness state is characterized by self-renewal and differentiation properties. However, stem cells are not able to preserve these characteristics in long-term culture because of the intrinsic fragility of their phenotype easily undergoing senescence or neoplastic transformation. Furthermore, although isolated from the same original tissue using similar protocols, adult stem cells can display dissimilar phenotypes and important cell clone/species contamination. Finally, the lack of a clear standardization contributes to complicate the comprehension about the stemness condition. In this context, cell lines displaying a particularly stable phenotype must be identified to define one or multiple benchmarks against which other stem cell lines could be reliably assessed. The present paper demonstrates that it is possible to isolate from the rat dental pulp a stem cell line (MUR-1) that does not display neoplastic transformation in long-term culture. MUR-1 cells stably express a broad range of stemness markers and are able to differentiate into adipogenic, osteogenic, chondrogenic, neurogenic, and cardiomyogenic lineages independently of the culture passages. Moreover, serial in vitro passages have not changed their immunophenotype, proliferation capacity, or differentiation potential. The uniqueness of these characteristics candidates MUR-1 as a model to reliably improve the understanding of the mechanisms governing the stem cell fate in the same as well as in other stem cell populations.
Stem cells and development 05/2012; · 4.15 Impact Factor
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ABSTRACT: Cardiac progenitor cells (CPCs) are a promising autologous source of cells for cardiac regenerative medicine. However, CPC culture in vitro requires the presence of microenvironmental conditions (a complex array of bioactive substance concentration, mechanostructural factors, and physicochemical factors) closely mimicking the natural cell surrounding in vivo, including the capability to uphold reactive oxygen species (ROS) within physiological levels in vitro. Cerium oxide nanoparticles (nanoceria) are redox-active and could represent a potent tool to control the oxidative stress in isolated CPCs. Here, we report that 24 h exposure to 5, 10, and 50 μg/mL of nanoceria did not affect cell growth and function in cardiac progenitor cells, while being able to protect CPCs from H(2)O(2)-induced cytotoxicity for at least 7 days, indicating that nanoceria in an effective antioxidant. Therefore, these findings confirm the great potential of nanoceria for controlling ROS-induced cell damage.
ACS Nano 04/2012; 6(5):3767-75. · 10.77 Impact Factor
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Giancarlo Forte,
Stefano Pietronave,
Giorgia Nardone,
Andrea Zamperone,
Eugenio Magnani,
Stefania Pagliari, Francesca Pagliari,
Cristina Giacinti,
Carmine Nicoletti,
Antonio Musaró,
Mauro Rinaldi,
Marco Ribezzo,
Chiara Comoglio,
Enrico Traversa,
Teruo Okano,
Marilena Minieri,
Maria Prat,
Paolo Di Nardo
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ABSTRACT: Human heart harbors a population of resident progenitor cells that can be isolated by stem cell antigen-1 antibody and expanded in culture. These cells can differentiate into cardiomyocytes in vitro and contribute to cardiac regeneration in vivo. However, when directly injected as single cell suspension, less than 1%-5% survive and differentiate. Among the major causes of this failure are the distressing protocols used to culture in vitro and implant progenitor cells into damaged hearts. Human cardiac progenitors obtained from the auricles of patients were cultured as scaffoldless engineered tissues fabricated using temperature-responsive surfaces. In the engineered tissue, progenitor cells established proper three-dimensional intercellular relationships and were embedded in self-produced extracellular matrix preserving their phenotype and multipotency in the absence of significant apoptosis. After engineered tissues were leant on visceral pericardium, a number of cells migrated into the murine myocardium and in the vascular walls, where they integrated in the respective textures. The study demonstrates the suitability of such an approach to deliver stem cells to the myocardium. Interestingly, the successful delivery of cells in murine healthy hearts suggests that myocardium displays a continued cell cupidity that is strictly regulated by the limited release of progenitor cells by the adopted source. When an unregulated cell source is added to the system, cells are delivered to the myocardium. The exploitation of this novel concept may pave the way to the setup of new protocols in cardiac cell therapy.
Stem Cells 12/2011; 29(12):2051-61. · 7.78 Impact Factor
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ABSTRACT: Cardiovascular diseases represent the main cause of morbidity and mortality worldwide. Millions of people are affected by such diseases in the industrialized countries, with hundreds of thousands new cases diagnosed every year. Among cardiac diseases, heart failure is the most common end-stage pathology, leading to impaired cardiac output and cardiac performance as a result of the irreversible loss of contractile cardiomyocytes. Tissue engineering holds the promise to provide personalized solutions to the problem of cardiac muscle repair. Indeed, the identification of little reservoirs of stem and progenitor cells within every body district opened new perspectives to the setup of patient-specific protocols for cardiac diseases. Nonetheless, the results of the first pre-clinical and clinical trials in which adult stem/progenitor cells were adopted pointed at the route of delivery to the injured organ as well as at the cell source as the main issues for cardiac tissue engineers. In fact, when adult stem cells were directly injected into the myocardium or delivered through bloodstream to the heart, no or few cells could be found engrafted within host tissue few days after the administration. Renewed enthusiasm was generated by the techniques set up to enrich cardiomyocytes obtained by embryonic stem cells and by the recent disclosure of the protocols to obtain reprogrammed pluripotent cells or reprogrammed cardiomyocytes out of patients' own somatic cells. In this context, additional efforts to setup efficient systems to deliver stem cells to the injured site are required. The application of forefront technologies to fabricate synthetic and hybrid scaffolds to be employed as cell delivery systems and the acknowledgement that surface physical, mechanical, chemical properties can exert specific effects on stem cells per se prompted new enthusiasm in the field. In this respect, a cardiac-specific scaffold should be able to comply with cardiac muscle architecture, be deformable as to indulge and possibly sustain cardiac contraction. As expected, such a scaffold should favor stem cell electromechanical coupling with host tissue, while promoting the vascularization of the newly-formed tissue.
Stem cell reviews 10/2011; · 5.08 Impact Factor
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08/2011; , ISBN: 978-953-307-688-1
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Stefania Pagliari,
Ana Cristina Vilela-Silva,
Giancarlo Forte, Francesca Pagliari,
Corrado Mandoli,
Giovanni Vozzi,
Stefano Pietronave,
Maria Prat,
Silvia Licoccia,
Arti Ahluwalia,
Enrico Traversa,
Marilena Minieri,
Paolo Di Nardo
Advanced Materials 01/2011; 23(4):514-8. · 13.88 Impact Factor
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Advanced Functional Materials 04/2010; 20(10):1617 - 1624. · 10.18 Impact Factor
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Corrado Mandoli,
Barbara Mecheri,
Giancarlo Forte, Francesca Pagliari,
Stefania Pagliari,
Felicia Carotenuto,
Roberta Fiaccavento,
Antonio Rinaldi,
Paolo Di Nardo,
Silvia Licoccia,
Enrico Traversa
Macromolecular Bioscience 02/2010; 10(2). · 3.89 Impact Factor
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Corrado Mandoli,
Barbara Mecheri,
Giancarlo Forte, Francesca Pagliari,
Stefania Pagliari,
Felicia Carotenuto,
Roberta Fiaccavento,
Antonio Rinaldi,
Paolo Di Nardo,
Silvia Licoccia,
Enrico Traversa
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ABSTRACT: The lack of a vascular network and poor perfusion is what mostly prevents three-dimensional (3D) scaffolds from being used in organ repair when reconstruction of thick tissues is needed. Highly-porous scaffolds made of poly(L-lactic acid) (PLLA) are prepared by directional thermally induced phase separation (dTIPS) starting from 1,4-dioxane/PLLA solutions. The influence of polymer concentration and temperature gradient, in terms of imposed intensity and direction, on pore size and distribution is studied by comparison with scaffolds prepared by isotropic TIPS. The processing parameters are optimized to achieve an overall porosity for the 3D scaffolds of about 93% with a degree of interconnectivity of 91%. The resulting pore network is characterized by the ordered repetition of closely packed dendrite-like cavities, each one showing stacks of 20 microm large side lamellar branches departing from 70 microm diameter vertical backbones, strongly resembling the vascular patterns. The in vitro biological responses after 1 and 2 weeks are evaluated from mesenchymal (bone marrow stromal) cells (MSC) static culturing. A novel vacuum-based deep-seeding method is set up to improve uniform cell penetration down to scaffold thicknesses of over 1 mm. Biological screenings show significant 3D scaffold colonization even after 18 h, while cellular retention is observed up to 14 d in vitro (DIV). Pore architecture-driven cellular growth is accompanied by cell tendency to preserve their multi-potency towards differentiation. Confluent tissues as thick as 1 mm were reconstructed taking advantage of the large perfusion enhanced by the highly porous microstructure of the engineered scaffolds, which could successfully serve for applications aimed at vascular nets and angiogenesis.
Macromolecular Bioscience 11/2009; 10(2):127-38. · 3.89 Impact Factor
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Sherif Soliman,
Stefania Pagliari,
Antonio Rinaldi,
Giancarlo Forte,
Roberta Fiaccavento, Francesca Pagliari,
Ornella Franzese,
Marilena Minieri,
Paolo Di Nardo,
Silvia Licoccia,
Enrico Traversa
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ABSTRACT: A novel (scalable) electrospinning process was developed to fabricate bio-inspired multiscale three-dimensional scaffolds endowed with a controlled multimodal distribution of fiber diameters and geared towards soft tissue engineering. The resulting materials finely mingle nano- and microscale fibers together, rather than simply juxtaposing them, as is commonly found in the literature. A detailed proof of concept study was conducted on a simpler bimodal poly(epsilon-caprolactone) (PCL) scaffold with modes of fiber distribution at 600 nm and 3.3 microm. Three conventional unimodal scaffolds with mean diameters of 300 nm and 2.6 and 5.2 microm, respectively, were used as controls to evaluate the new materials. Characterization of the microstructure (i.e. porosity, fiber distribution and pore structure) and mechanical properties (i.e. stiffness, strength and failure mode) indicated that the multimodal scaffold had superior mechanical properties (Young's modulus approximately 40MPa and strength approximately 1MPa) in comparison with the controls, despite the large porosity ( approximately 90% on average). A biological assessment was conducted with bone marrow stromal cell type (mesenchymal stem cells, mTERT-MSCs). While the new material compared favorably with the controls with respect to cell viability (on the outer surface), it outperformed them in terms of cell colonization within the scaffold. The latter result, which could neither be practically achieved in the controls nor expected based on current models of pore size distribution, demonstrated the greater openness of the pore structure of the bimodal material, which remarkably did not come at the expense of its mechanical properties. Furthermore, nanofibers were seen to form a nanoweb bridging across neighboring microfibers, which boosted cell motility and survival. Lastly, standard adipogenic and osteogenic differentiation tests served to demonstrate that the new scaffold did not hinder the multilineage potential of stem cells.
Acta biomaterialia 10/2009; 6(4):1227-37. · 3.98 Impact Factor
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Giancarlo Forte,
Felicia Carotenuto, Francesca Pagliari,
Stefania Pagliari,
Paolo Cossa,
Roberta Fiaccavento,
Arti Ahluwalia,
Giovanni Vozzi,
Bruna Vinci,
Annalucia Serafino,
Antonio Rinaldi,
Enrico Traversa,
Luciana Carosella,
Marilena Minieri,
Paolo Di Nardo
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ABSTRACT: The replacement of injured cardiac contractile cells with stem cell-derived functionally efficient cardiomyocytes has been envisaged as the resolutive treatment for degenerative heart diseases. Nevertheless, many technical issues concerning the optimal procedures to differentiate and engraft stem cells remain to be answered before heart cell therapy could be routinely used in clinical practice. So far, most studies have been focused on evaluating the differentiative potential of different growth factors without considering that only the synergistic cooperation of biochemical, topographic, chemical, and physical factors could induce stem cells to adopt the desired phenotype. The present study demonstrates that the differentiation of cardiac progenitor cells to cardiomyocytes does not occur when cells are challenged with soluble growth factors alone, but requires strictly controlled procedures for the isolation of a progenitor cell population and the artifactual recreation of a microenvironment critically featured by a fine-tuned combination of specific biological and physical factors. Indeed, the scaffold geometry and stiffness are crucial in enhancing growth factor differentiative effects on progenitor cells. The exploitation of this concept could be essential in setting up suitable procedures to fabricate functionally efficient engineered tissues. Disclosure of potential conflicts of interest is found at the end of this article.
Stem Cells 06/2008; 26(8):2093-103. · 7.78 Impact Factor
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Giancarlo Forte,
Felicia Carotenuto, Francesca Pagliari,
Stefania Pagliari,
Paolo Cossa,
Roberta Fiaccavento,
Arti Ahluwalia,
Giovanni Vozzi,
Bruna Vinci,
Annalucia Serafino,
Antonio Rinaldi,
Enrico Traversa,
Luciana Carosella,
Marilena Minieri,
Paolo Di Nardo M.D
[show abstract]
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ABSTRACT: The replacement of injured cardiac contractile cells with stem cell-derived functionally efficient cardiomyocytes has been envisaged as the resolutive treatment for degenerative heart diseases. Nevertheless, many technical issues concerning the optimal procedures to differentiate and engraft stem cells remain to be answered before heart cell therapy could be routinely used in clinical practice. So far, most studies have been focused on evaluating the differentiative potential of different growth factors without considering that only the synergistic cooperation of biochemical, topographic, chemical, and physical factors could induce stem cells to adopt the desired phenotype. The present study demonstrates that the differentiation of cardiac progenitor cells to cardiomyocytes does not occur when cells are challenged with soluble growth factors alone, but requires strictly controlled procedures for the isolation of a progenitor cell population and the artifactual recreation of a microenvironment critically featured by a fine-tuned combination of specific biological and physical factors. Indeed, the scaffold geometry and stiffness are crucial in enhancing growth factor differentiative effects on progenitor cells. The exploitation of this concept could be essential in setting up suitable procedures to fabricate functionally efficient engineered tissues.Disclosure of potential conflicts of interest is found at the end of this article.
Stem Cells 05/2008; 26(8):2093 - 2103. · 7.78 Impact Factor
