Browsing by Author "Bártolo, Paulo J."
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- 3D printing of new biobased unsaturated polyesters by microstereo-thermal-lithographyPublication . Gonçalves, Filipa A. M. M.; Costa, Cátia S. M. F.; Fabela, Inês G. P.; Farinha, Dina; Faneca, Henrique; Simões, Pedro N.; Serra, Arménio C.; Bártolo, Paulo J.; Coelho, Jorge F. J.New micro three-dimensional (3D) scaffolds using biobased unsaturated polyesters (UPs) were prepared by microstereo-thermal-lithography (μSTLG). This advanced processing technique offers indubitable advantages over traditional printing methods. The accuracy and roughness of the 3D structures were evaluated by scanning electron microscopy and infinite focus microscopy, revealing a suitable roughness for cell attachment. UPs were synthesized by bulk polycondensation between biobased aliphatic diacids (succinic, adipic and sebacic acid) and two different glycols (propylene glycol and diethylene glycol) using fumaric acid as the source of double bonds. The chemical structures of the new oligomers were confirmed by proton nuclear magnetic resonance spectra, attenuated total reflectance Fourier transform infrared spectroscopy and matrix assisted laser desorption/ionization-time of flight mass spectrometry. The thermal and mechanical properties of the UPs were evaluated to determine the influence of the diacid/glycol ratio and the type of diacid in the polyester's properties. In addition an extensive thermal characterization of the polyesters is reported. The data presented in this work opens the possibility for the use of biobased polyesters in additive manufacturing technologies as a route to prepare biodegradable tailor made scaffolds that have potential applications in a tissue engineering area.
- Cell-instructive pectin hydrogels crosslinked via thiol-norbornene photo-click chemistry for skin tissue engineeringPublication . Pereira, Rúben; Barrias, Cristina C.; Bártolo, Paulo J.; Granja, Pedro L.Cell-instructive hydrogels are attractive for skin repair and regeneration, serving as interactive matrices to promote cell adhesion, cell-driven remodeling and de novo deposition of extracellular matrix compo nents. This paper describes the synthesis and photocrosslinking of cell-instructive pectin hydrogels using cell-degradable peptide crosslinkers and integrin-specific adhesive ligands. Protease-degradable hydro gels obtained by photoinitiated thiol-norbornene click chemistry are rapidly formed in the presence of dermal fibroblasts, exhibit tunable properties and are capable of modulating the behavior of embedded cells, including the cell spreading, hydrogel contraction and secretion of matrix metalloproteases. Keratinocytes seeded on top of fibroblast-loaded hydrogels are able to adhere and form a compact and dense layer of epidermis, mimicking the architecture of the native skin. Thiol-ene photocrosslinkable pec tin hydrogels support the in vitro formation of full-thickness skin and are thus a highly promising plat form for skin tissue engineering applications, including wound healing and in vitro testing mod
- Electrospun polycaprolactone (PCL) degradation: An in vitro and in vivo studyPublication . Dias, Juliana R.; Sousa, Aureliana; Augusto, Ana; Bártolo, Paulo J.; Granja, Pedro L.Polycaprolactone (PCL) is widely used in tissue engineering due to its interesting properties, namely biocompatibility, biodegradability, elastic nature, availability, cost efficacy, and the approval of health authorities such as the American Food and Drug Administration (FDA). The PCL degradation rate is not the most adequate for specific applications such as skin regeneration due to the hydrophobic nature of bulk PCL. However, PCL electrospun fiber meshes, due to their low diameters resulting in high surface area, are expected to exhibit a fast degradation rate. In this work, in vitro and in vivo degradation studies were performed over 90 days to evaluate the potential of electrospun PCL as a wound dressing. Enzymatic and hydrolytic degradation studies in vitro, performed in a static medium, demonstrated the influence of lipase, which promoted a rate of degradation of 97% for PCL meshes. In an in vivo scenario, the degradation was slower, although the samples were not rejected, and were well-integrated in the surrounding tissues inside the subcutaneous pockets specifically created.
- Extruded bioreactor perfusion culture supports the chondrogenic differentiation of human mesenchymal stem/stromal cells in 3D porous poly(ɛ-caprolactone) scaffoldsPublication . Silva, João C.; Moura, Carla; Borrecho, Gonçalo; Matos, António P. Alves de; Silva, Cláudia L. da; Cabral, Joaquim M. S.; Bártolo, Paulo J.; Linhardt, Robert J.; Ferreira, Frederico CasteloNovel bioengineering strategies for the ex vivo fabrication of native-like tissue-engineered cartilage are crucial for the translation of these approaches to clinically manage highly prevalent and debilitating joint diseases. Bioreactors that provide different biophysical stimuli have been used in tissue engineering approaches aimed at enhancing the quality of the cartilage tissue generated. However, such systems are often highly complex, expensive, and not very versatile. In the current study, a novel, cost-effective, and customizable perfusion bioreactor totally fabricated by additive manufacturing (AM) is proposed for the study of the effect of fluid flow on the chondrogenic differentiation of human bone-marrow mesenchymal stem/stromal cells (hBMSCs) in 3D porous poly(ɛ-caprolactone) (PCL) scaffolds. hBMSCs are first seeded and grown on PCL scaffolds and hBMSC–PCL constructs are then transferred to 3D-extruded bioreactors for continuous perfusion culture under chondrogenic inductive conditions. Perfused constructs show similar cell metabolic activity and significantly higher sulfated glycosaminoglycan production (≈1.8-fold) in comparison to their non-perfused counterparts. Importantly, perfusion bioreactor culture significantly promoted the expression of chondrogenic marker genes while downregulating hypertrophy. This work highlights the potential of customizable AM platforms for the development of novel personalized repair strategies and more reliable in vitro models with a wide range of applications.
- Numerical Calculations in Tissue EngineeringPublication . Almeida, Henrique de Amorim; Bártolo, Paulo J.The design of optimized scaffolds for tissue engineering is a key topic of research, as the complex macro- and micro- architectures required for a scaffold depends not only on the mechanical properties, but also on the physical and molecular queues of the surrounding tissue within the defect site. Thus, the prediction of optimal features for tissue engineering scaffolds is very important for its mechanical, vascular or topological properties. The relationship between high scaffold porosity and high mechanical properties is contradictory, as it becomes even more complex due to the scaffold degradation process. A scaffold design strategy was developed, based on the finite element method, to optimise the scaffold design regarding the mechanical and vascular properties as a function of porosity. Scaffolds can be considered as a LEGO structure formed by an association of small elementary units or blocks. In this research work, two types of family elementary scaffold units were considered: non-triple periodic minimal surfaces and triple periodic minimal surfaces that describe natural existing surfaces. The main objective of this work is to present the undergoing research based on numerical simulations for the evaluation and prediction of the scaffold's behaviour under structural and vascular loading, and its topological optimisation.
- Photocrosslinkable Materials for the Fabrication of Tissue-Engineered Constructs by StereolithographyPublication . Pereira, Rúben F.; Bártolo, Paulo J.Stereolithography is an additive technique that produces three-dimensional (3D) solid objects using a multi-layer procedure through the selective photoinitiated curing reaction of a liquid photosensitive material. Stereolithographic processes have been widely employed in Tissue Engineering for the fabrication of temporary constructs, using natural and synthetic polymers, and polymer-ceramic composites. These processes allow the fabrication of complex structures with a high accuracy and precision at physiological temperatures, incorporating cells and growth factors without significant damage or denaturation. Despite recent advances on the development of novel biomaterials and biocompatible crosslinking agents, the main limitation of these techniques are the lack number of available photocrosslinkable materials, exhibiting appropriate biocompatibility and biodegradability. This chapter gives an overview of the current state-of-art of materials and stereolithographic techniques to produce constructs for tissue regeneration, outlining challenges for future research.
- A single-component hydrogel bioink for bioprinting of bioengineered 3D constructs for dermal tissue engineeringPublication . Pereira, Rúben; Sousa, Aureliana; Barrias, Cristina C.; Bártolo, Paulo J.; Granja, Pedro L.Bioprinting is attractive to create cellularized constructs for skin repair. However, the vast majority of bioinks present limitations in the printing of chemically defined 3D constructs with controllable biophysical and biochemical properties. To address this challenge, a single-component hydrogel bioink with a controlled density of cell-adhesive ligands, tuneable mechanical properties and adjustable rheological behaviour is developed for extrusion bioprinting and applied for the biofabrication of 3D dermal constructs. A methacrylate modified pectin bioink is designed to allow the tethering of integrin-binding motifs and the formation of hydrogels by UV photopolymerization and ionic gelation. The rheological behaviour of a low polymer concentration (1.5 wt%) solution is adjusted by ionic crosslinking, yielding a printable bioink that holds the predesigned shape upon deposition for postprinting photocrosslinking. Printed constructs provide a suitablemicroenvironment that supports the deposition of endogenous extracellular matrix, rich in collagen and fibronectin, by entrapped dermal fibroblasts. This approach enables the design of chemically defined and cell-responsive bioinks for tissue engineering applications and particularly for the generation of biomimetic skin constructs.
- Sustainability in extrusion-based additive manufacturing technologiesPublication . Freitas, Dino; Henrique A. Almeida; Bartolo, Helena; Bártolo, Paulo J.Growing environmental and social concerns about human society’s impact on the natural environment have been pushing sustainable development issues into the front line of public policies. The degradation of natural resources has been aggravated by several other factors: the actual lifestyle of consumer societies, the rapid growth of emerging countries, rising inequalities among regions, and the reduction of the life cycle time for each product requiring a high consumption of limited resources. Sustainable industrial practices can contribute to the development of more sustainable products and processes. It is critical to apply eco-design principles and develop greener products and production processes, reducing impacts associated with its production and consumption. An environmental impact assessment should be taken into account from the early design and production phases, through all the stages of a product’s life until disposal. As public interest in additive manufacturing grows, its increasing usage for the production of final parts can support a drive towards more sustainable manufacturing processes. To introduce sustainability awareness during extrusion-based additive manufacturing productions, a study of the production of a part in terms its building orientation and internal filling, regarding the production time, energy consumption and end-of-life scenarios is considered. By combining the additive processes with eco-design, a higher awareness is possible, which results in added value to the produced parts while maintaining the part’s objective performance.
- Traditional Therapies for Skin Wound HealingPublication . Pereira, Rúben F.; Bártolo, Paulo J.Significance: The regeneration of healthy and functional skin remains a huge challenge due to its multilayer structure and the presence of different cell types within the extracellular matrix in an organized way. Despite recent advances in wound care products, traditional therapies based on natural origin compounds, such as plant extracts, honey, and larvae, are interesting alternatives. These therapies offer new possibilities for the treatment of skin diseases, enhancing the access to the healthcare, and allowing overcoming some limitations associated to the modern products and therapies, such as the high costs, the long manufacturing times, and the increase in the bacterial resistance. This article gives a general overview about the recent advances in traditional therapies for skin wound healing, focusing on the therapeutic activity, action mechanisms, and clinical trials of the most commonly used natural compounds. New insights in the combination of traditional products with modern treatments and future challenges in the field are also highlighted. Recent Advances: Natural compounds have been used in skin wound care for many years due to their therapeutic activities, including anti-inflammatory, antimicrobial, and cell-stimulating properties. The clinical efficacy of these compounds has been investigated through in vitro and in vivo trials using both animal models and humans. Besides the important progress regarding the development of novel extraction methods, purification procedures, quality control assessment, and treatment protocols, the exact mechanisms of action, side effects, and safety of these compounds need further research. Critical Issues: The repair of skin lesions is one of the most complex biological processes in humans, occurring throughout an orchestrated cascade of overlapping biochemical and cellular events. To stimulate the regeneration process and prevent the wound to fail the healing, traditional therapies and natural products have been used with promising results. Although these products are in general less expensive than the modern treatments, they can be sensitive to the geographic location and season, and exhibit batch-to-batch variation, which can lead to unexpected allergic reactions, side effects, and contradictory clinical results. Future Directions: The scientific evidence for the use of traditional therapies in wound healing indicates beneficial effects in the treatment of different lesions. However, specific challenges remain unsolved. To extend the efficacy and the usage of natural substances inwound care,multidisciplinary efforts are necessary to prove the safety of these products, investigate their side effects, and develop standard controlled trials. The development of good manufacturing practices and regulatory legislation also assume a pivotal role in order to improve the use of traditional therapies by the clinicians and to promote their integration into the national health system. Current trends move to the development of innovative wound care treatments, combining the use of traditional healing agents and modern products/practices, such as nanofibers containing silver nanoparticles,Aloe veraloaded into alginate hydrogels, propolis into dressing films, and hydrogel sheets containing honey.