Percorrer por autor "Bártolo, P. J."
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- Adhesion, proliferation and distribution of human mesenchymal stem/stromal cells (MSCs) in Poly(ɛ -caprolactone) (PCL) scaffolds with different pore sizesPublication . Moura, C. S.; Biscaia, S.; Viana, T.; Bártolo, P. J.; Silva, C. L. da; Cabral, J. S.; Ferreira, F. C.Tissue engineering, combining the use of biomaterials, mesenchymal stem/stromal cells (MSCs) and optimized culture medium formulations, is a very promising research field for tissue regeneration. This work is focused on the application of Poly(ε-caprolactone) scaffolds designed to have a spacing gradient between orthogonal fibres with diameters of 0.3mm with pore sizes ranging from 190 to 390 μm. Human bone marrow (BM)-derived MSCs, were used to evaluate cell adhesion, proliferation and distribution within the scaffolds. Specifically, we hypothesized if seeding MSC in scaffold regions with different pore sizes would influence cell proliferation. Therefore, MSC were seeded at the centre of scaffolds with either larger (390 μm) or smaller pores (190 μm). Results obtained showed similar levels of adhesion and proliferation in both configurations, with fold increase in total cell number of 5 after 13 days of cultivation; which indicates that no limitation for cell proliferation within the range of pore sizes studied is observed.
- Analysis of manufacturing parameters on the shear strength of aluminium adhesive single-lap jointsPublication . Pereira, A. M.; Ferreira, J. M.; Antunes, F. V.; Bártolo, P. J.An experimental and numerical investigation into the shear strength behaviour of aluminium alloy adhesive lap joints was carried out in order to understand the effect of geometrical and manufacturing parameters on the strength of adhesive bonding joints, with the aim of optimizing shear strength. The adherend material used for the experimental tests was an aluminium alloy in the form of thin sheets, and the adhesive used was a high strength epoxy. Five surface treatments were studied. The surface treatments process using sodium dichromate-sulphuric acid etch (CSA) and abrasive polishing (AP) resulted in improved joint shear strength when compared to acetone cleaning (SW), caustic etch (CE), and Tucker's reagent etch (TR). The decrease in surface roughness was found to increase the shear strength of single-lap joints. An increase in adherend thickness and overlap length was found to increase shear strength which means that an increase in joint rigidity increases its strength. A numerical analysis was developed to explain the effect of the geometrical parameters on rotation angle, stress and strain fields, and failure load. An increase in adherend thickness and overlap length decreases the joint rotation angle, reducing the plastic strain peak and therefore increasing the failure load.
- Biomechanical performance of hybrid electrospun structures for skin regenerationPublication . Dias, J.R.; Baptista-Silva, S.; Sousa, A.; Oliveira, A. L.; Bártolo, P. J.; Granja, P. L.Wound dressings made by electrospun nanofibers have been demonstrating great potential to regenerate skin tissue as compared to the conventional membrane products available in the market. Until today most of the developed dressings have only demonstrated the capability to regenerate the dermis or epidermis. In this study we propose new hybrid electrospun meshes combining polycaprolactone and gelatin. Several approaches, multilayer, coating and blend were stablished to investigate the most appropriate hybrid structure with potential to promote skin regeneration in its full thickness. The structures were evaluated in terms of physico-chemical properties (porosity, water vapor permeability, contact angle and swelling degree) and according to its mechanical and biological performance. Multilayer and blend structures demonstrated to fit most of native skin requirements. However, looking to all the performed characterization we considered multilayer as the most promising hybrid structures, due its high porosity which contributed to an ideal water vapor permeability rate and good mechanical and biological properties. Based on this multilayer structure is a promisor wound dressing.
- Computational technologies in tissue engineeringPublication . Almeida, H. A.; Bártolo, P. J.In last decades, many advances have been made in order to aid the medical community. Numerous computational technologies have been developed and improved the efficiency in diagnostic and treatment of many diseases. Many of the technologies were developed with the main goal of aiding in the research of genetic and viral diseases. Tissue engineering is a multidisciplinary field that requires the combined effort of cell biologists, engineers, material scientists, mathematicians, geneticists, and clinicians toward the development of biological substitutes that restore, maintain, or improve tissue function. The success of this emerging medical domain relies on the current technological advances. This paper presents an overview of the existing computational technologies that have been implemented in tissue engineering and the design of scaffolds for tissue engineering applications. These computational technologies contemplate medical imaging processing, numerical calculations (structural, vascular and topological) and biofabrication techniques necessary for the scaffolds optimum design and production.
- Effect of process parameters on the strength of resistance spot welds in 6082-T6 aluminium alloyPublication . Pereira, A. M.; Ferreira, J. M.; Loureiro, A.; Costa, J. D. M.; Bártolo, P. J.In this study the microstructural and mechanical behaviour of resistance spot welds (RSW) done on aluminium alloy 6082-T6 sheets, welded at different welding parameters, is examined. Microstructural examinations and hardness evaluations were carried out in order to determine the influence of welding parameters on the quality of the welds. The welded joints were subjected to static tensile-shear tests in order to determine their strength and failure mode. The increase in weld current and duration increased the nugget size and the weld strength. Beyond a critical nugget diameter the failure mode changed from interfacial to pullout. Taking into consideration the sheet thickness and the mechanical properties of the weld, a simple model is proposed to predict the critical nugget diameter required to produce pull-out failure mode in undermatched welds in heat-treatable aluminium alloys.
- Improved osteoblast cell affinity on plasma-modified 3-D extruded PCL scaffoldsPublication . Domingos, M.; Intranuovo, F.; Gloria, A.; Gristina, R.; Ambrosio, L.; Bártolo, P. J.; Favia, P.Cellular adhesion and proliferation inside three-dimensional synthetic scaffolds represent a major challenge in tissue engineering. Besides the surface chemistry of the polymers, it is well recognized that scaffold internal architecture, namely pore size/shape and interconnectivity, has a strong effect on the biological response of cells. This study reports for the first time how polycaprolactone (PCL) scaffolds with controlled micro-architecture can be effectively produced via bioextrusion and used to enhance the penetration of plasma deposited species. Low-pressure nitrogen-based coatings were employed to augment cell adhesion and proliferation without altering the mechanical properties of the structures. X-ray photoelectron spectroscopy carried out on different sections of the scaffolds indicates a uniform distribution of nitrogen-containing groups throughout the entire porous structure. In vitro biological assays confirm that plasma deposition sensitively promotes the activity of Saos-2 osteoblast cells, leading to a homogeneous colonization of the PCL scaffolds.
- In situ crosslinked electrospun gelatin nanofibers for skin regenerationPublication . Dias, J. R.; Baptista-Silva, S.; Oliveira, C. M. T. de; Sousa, A.; Oliveira, A. L.; Bártolo, P. J.; Granja, P. L.Due to its intrinsic similarity to the extracellular matrix, gelatin electrospun nanofibrous meshes are promising scaffold structures for wound dressings and tissue engineering applications. However, gelatin is water soluble and presents poor mechanical properties, which generally constitute relevant limitations to its applicability. In this work, gelatin was in situ crosslinked with 1,4-butanediol diglycidyl ether (BDDGE) at different concentrations (2, 4 and 6 wt%) and incubation time-points (24, 48 and 72 h) at 37 °C. The physico-chemical and biological properties of BDDGE-crosslinked electrospun gelatin meshes were investigated. Results show that by changing the BDDGE concentration it is possible to produce nanofibers crosslinked in situ with well-defined morphology and modulate fiber size and mechanical properties. Crosslinked gelatin meshes show no toxicity towards fibroblasts, stimulating their adhesion, proliferation and synthesis of new extracellular matrix, thereby indicating the potential of this strategy for skin tissue engineering.
- Mesenchymal Stem Cells and Biomaterials Systems – Perspectives for Skeletal Muscle Tissue Repair and RegenerationPublication . Caseiro, A.R.; Pereira, T.; Bártolo, P. J.; Santos, J.D.; Luís, A.L.; Maurício, A.C.Skeletal muscle is essential in voluntary movement and other major vital functions. Muscle injuries are important in clinical practice and, despite skeletal muscle's good regenerative ability, severe tissue loss impairs complete myofibre regeneration, limiting structural and functional recovery of the affected muscle, eventually leading to the development of non-contractile fibrous scar. The intrinsic healing mechanisms rely in great extent on the residing progenitor population but significant drawbacks to their practical application in regenerative strategies boosted the search for alternative cell sources, such as extra-fetal mesenchymal stem cells (MSCs). MSCs have demonstrated to positively influence the regeneration of different disease models. When severe volumetric muscle tissue loss occurs, the body is seldom capable of replacing the lost portions with fully functional tissue. A rational strategy to aid the healing of such situations is the application of biomaterial implants that provide a structural matrix for the ingrowth of regenerating muscle fibres. Both synthetic and natural biomaterials have been hypothesized for this purpose and some have reached as far clinical cases applications. Obvious improvements are observed in most cases, but reaction to some biomaterials and functional recovery are still a challenge. The addition of MSCs to the biomaterials seems to improve the systems' performance in the overall regenerative milieu. This strategies promote scaffold's vascularization and integration, as well as accelerated tissue ingrowth and reduces scar formation, resulting in improved recovery rates at both structural and functional levels.
- Permeability Evaluation of Flow Behaviors Within Perfusion BioreactorsPublication . Freitas, D.; Almeida, H.A.; Bártolo, P. J.Tissue engineering aims to produce artificial tissue in order to create or repair damaged tissue. It is evident that scaffolds are of extreme importance, because they will be the support structure of the new tissue. This new tissue is cultivated in vitro in a bioreactor in which is placed the scaffold. In order to control the cell culture process inside of a bioreactor, it is essential to know the fluid flow inside the scaffold for an adequate exchange of nutrients and metabolic waste. A novel multifunctional bioreactor with a perfusion system module comprised of three different inlet and outlet membranes is being developed. This research work will evaluate the permeability of the scaffold under the three different inlet and outlet diffusion membranes of the culture chamber.
- Promoting Nerve Regeneration in a Neurotmesis Rat Model Using Poly(DL-lactide-ε-caprolactone) Membranes and Mesenchymal Stem Cells from the Wharton’s Jelly: In Vitro and In Vivo AnalysisPublication . Pereira, T.; Gärtner, A.; Amorim, I.; Almeida, A.; Caseiro, A.R.; Armada-da-Silva, Paulo A. S.; Amado, Sandra; Fregnan, Federica; Varejão, A. S. P.; Santos, J. D.; Bártolo, P. J.; Geuna, S.; Luís, A. L.; Maurício, A. C.In peripheral nerves MSCs can modulate Wallerian degeneration and the overall regenerative response by acting through paracrine mechanisms directly on regenerating axons or upon the nerve-supporting Schwann cells. In the present study, the effect of human MSCs from Wharton’s jelly (HMSCs), differentiated into neuroglial-like cells associated to poly (DL-lactide-ε-caprolactone) membrane, on nerve regeneration, was evaluated in the neurotmesis injury rat sciatic nerve model. Resultsin vitroshowed successful differentiation of HMSCs into neuroglial-like cells, characterized by expression of specific neuroglial markers confirmed by immunocytochemistry and by RT-PCR and qPCR targeting specific genes expressed.In vivotesting evaluated during the healing period of 20 weeks, showed no evident positive effect of HMSCs or neuroglial-like cell enrichment at the sciatic nerve repair site on most of the functional and nerve morphometric predictors of nerve regeneration although the nociception function was almost normal. EPT on the other hand, recovered significantly better after HMSCs enriched membrane employment, to values of residual functional impairment compared to other treated groups. When the neurotmesis injury can be surgically reconstructed with an end-to-end suture or by grafting, the addition of a PLC membrane associated with HMSCs seems to bring significant advantage, especially concerning the motor function recovery.
