1,984 research outputs found
Adipose tissue derived stem cells secretome: soluble factors and their roles in regenerative medicine
Stem cells have been long looked at as possible therapeutic vehicles for different health related problems. Among the different
existing stem cell populations, Adipose derived Stem Cells (ASCs) have been gathering attention in the last 10 years. When compared to
other stem cells populations and sources, ASCs can be easily isolated while providing higher yields upon the processing of adipose tissue.
Similar to other stem cell populations, it was initially thought that the main potential of ASCs for regenerative medicine approaches was
intimately related to their differentiation capability. Although this is true, there has been an increasing body of literature describing the
trophic effects of ASCs on the protection, survival and differentiation of a variety of endogenous cells/tissues. Moreover, they have also
shown to possess an immunomodulatory character. This effect is closely related to the ASCsâ secretome and the soluble factors found
within it. Molecules such as hepatocyte growth factor (HGF), granulocyte and macrophage colony stimulating factors, interleukins (ILs)
6, 7, 8 and 11, tumor necrosis factor-α (TNF-α), vascular endothelial growth factor (VEGF), brain derived neurotrophic factor (BDNF),
nerve growth factor (NGF), adipokines and others have been identified within the ASCsâ secretome. Due to its importance regarding future
applications for the field of regenerative medicine, we aim, in the present review, to make a comprehensive analysis of the literature
relating to the ASCsâ secretome and its relevance to the immune and central nervous system, vascularization and cardiac regeneration.
The concluding section will highlight some of the major challenges that remain before ASCs can be used for future clinical applications
Encapsulation of human articular chondrocytes into 3D hydrogel : phenotype and genotype characterization
This chapter is intended to provide a summary of the current materials used in cell encapsulation technology
as well as methods for evaluating the performance of cells encapsulated in a polymeric matrix. In
particular, it describes the experimental procedure to prepare a hydrogel matrix based on natural polymers
for encapsulating and culturing human articular chondrocytes with the interest in cartilage regeneration.
Protocols to evaluate the viability, proliferation, differentiation, and matrix production of
embedded cells are also described and include standard protocols such as the MTT and [3H] Thymidine
assays, reverse transcription polymerase chain reaction (RT-PCR) technique, histology, and immunohistochemistry
analysis. The assessment of cell distribution within the 3D hydrogel construct is also described
using APoTome analysis.(undefined
The effect of insulin-loaded chitosan particle-aggregated scaffolds in chondrogenic differentiation
Osteochondral defect repair requires a tissue engineering approach that aims at mimicking the physiological
properties and structure of two different tissues (cartilage and bone) using a scaffoldâcell construct. One ideal
approach would be to engineer in vitro a hybrid material using a single-cell source. For that purpose, the scaffold
should be able to provide the adequate biochemical cues to promote the selective but simultaneous differentiation
of both tissues. In this work, attention was paid primarily to the chondrogenic differentiation by focusing
on the development of polymeric systems that provide biomolecules release to induce chondrogenic differentiation.
For that, different formulations of insulin-loaded chitosan particleâaggregated scaffolds were developed
as a potential model system for cartilage and osteochondral tissue engineering applications using insulin as a
potent bioactive substance known to induce chondrogenic differentiation. The insulin encapsulation efficiency
was shown to be high with values of 70.37!0.8%, 84.26!1.76%, and 87.23!1.58% for loadings of 0.05%, 0.5%,
and 5%, respectively. The in vitro release profiles were assessed in physiological conditions mimicking the cell
culture procedures and quantified by Micro-BCA! protein assay. Different release profiles were obtained that
showed to be dependent on the initial insulin-loading percentage. Further, the effect on prechondrogenic
ATDC5 cells was investigated for periods up to 4 weeks by studying the influence of these release systems on
cell morphology, DNA and glycosaminoglycan content, histology, and gene expression of collagen types I and II,
Sox-9, and aggrecan assessed by real-time polymerase chain reaction. When compared with control conditions
(unloaded scaffolds cultured with the standard chondrogenic-inducing medium), insulin-loaded scaffolds upregulated
the Sox-9 and aggrecan expression after 4 weeks of culture. From the overall results, it is reasonable to
conclude that the developed loaded scaffolds when seeded with ATDC5 can provide biochemical cues for
chondrogenic differentiation. Among the tested formulations, the higher insulin-loaded system (5%) was the
most effective in promoting chondrogenic differentiation.The authors would like to acknowledge the Portuguese Foundation for Science and Technology for the Ph. D. Grant to Patricia B. Malafaya (SFRH/BD/11155/2002). This work was partially supported and carried out under the scope of the European STREP Project HIPPOCRATES (NMP3-CT-2003-505758) and European NoE EXPERTISSUES (NMP3CT-2004-500283). The authors also like to acknowledge the Life and Health Sciences Research Institute (ICVS), University of Minho, for the use of their facilities, namely, to Luis Martins for histological sections slicing and H&E stain processing
Subcritical carbon dioxide foaming of polycaprolactone for bone tissue regeneration
Accepted manuscriptThe preparation of three-dimensional polycaprolactone scaffolds using dense CO2 as foaming agent, without supercritical conditions, was evaluated in this study towards future applications in bone repair. Herein, 3D foams were obtained at 5.0âŻMPa and 45âŻÂ°C. To induce bioactivity, ÎČ-tricalcium phosphate (ÎČ-TCP, 10âŻwt%) and dexamethasone (5 and 10âŻwt%) were dispersed in the scaffolds. Foams revealed a pore size range of 164â882âŻÎŒm, 73â99% porosity and 79â99% interconnectivity, assessed by micro-computed tomography, and a Young modulus of 1.76â2.92âŻMPa. Dexamethasone did not impair morphology of the matrices in comparison with PCL+ÎČ-TCP, which presented a water uptake of nearly 100% after 14 days. A sustained release of dexamethasone was achieved over 35 days in physiologic solution. This study reports the feasibility of using dense CO2 to produce in one-step a porous matrix loaded with active agents opening new possibilities towards injectable systems for in situ foamingEuropean Union Seventh Framework Programme (FP7/2007-2013) under grant agreement number REGPOT-CT2012-316331-POLARIS. It was also funded by the project âNovel smart and biomimetic materials for innovative regenerative medicine approachesâ (RL1-ABMR-NORTE-01-0124-FEDER-000016) co-financed by North Portugal Regional Operational Programme (ON.2 â O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF) and the project NORTEâ01â0145âFEDERâ000013, supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement. The authors would like to acknowledge the funding of the project Associate Laboratory ICVS/3Bâs, under grant agreement number POCI-01-0145-FEDER-007038 supported by FEDER, through the Competitiveness Factors Operational Programme (COMPETE), and by National funds, through the Foundation for Science and Technology (FCT)info:eu-repo/semantics/publishedVersio
Biomaterials as Tendon and Ligament Substitutes: Current Developments
Tendon and ligament have specialized dynamic microenvironment characterized by a complex hierarchical extracellular matrix essential for tissue functionality, and responsible to be an instructive niche for resident cells. Among musculoskeletal diseases, tendon/ligament injuries often result in pain, substantial tissue morbidity, and disability, affecting athletes, active working people and elder population. This represents not only a major healthcare problem but it implies considerable social and economic hurdles. Current treatments are based on the replacement and/or augmentation of the damaged tissue with severe associated limitations. Thus, it is evident the clinical challenge and emergent need to recreate native tissue featuresĂÂ and regenerate damaged tissues. In this context, the design and development of anisotropic bioengineered systems with potential to recapitulate the hierarchical architecture and organization of tendons and ligaments from nano to macro scale will be discussed in this chapter. Special attention will be given to the state-of-the-art fabrication techniques, namely spinning and electrochemical alignment techniques to address the demanding requirements for tendon substitutes, particularly concerning the importance of biomechanical and structural cues of these tissues. Moreover, the poor innate regeneration ability related to the low cellularity and vascularization of tendons and ligaments also anticipates the importance of cell based strategies, particularly on the stem cells role for the success of tissue engineered therapies. In summary, this chapter provides a general overview on tendon and ligaments physiology and current conventional treatments for injuries caused by trauma and/or disease. Moreover, this chapter presents tissue engineering approaches as an alternative to overcome the limitations of current therapies, focusing on the discussion about scaffolds design for tissue substitutes to meet the regenerative medicine challenges towards the functional restoration of damaged or degenerated tendon and ligament tissues.Portuguese Foundation for Science and Technology for the post-doctoral grant (SFRH/BPD/111729/2015) and for the projects Recognize (UTAP-ICDT/CTM-BIO/0023/2014) and POC
I-01-0145-FEDER-007
Mesenchymal stem cells in the umbilical cord: phenotypic characterization, secretome and applications in central nervous system regenerative medicine
Mesenchymal Stem Cells (MSCs), have been defined and characterized by: 1) their ability to adhere to plastic culture flasks; 2) the positive expression of CD105, CD73, CD90 membrane antigens, and the lack of expression of others (e.g CD45 and CD34) and 3) the ability of differentiation under adequate conditions along the osteogenic, chondrogenic and adipogenic lineages. In recent years, cells with these characteristics have been isolated from the Whartonâs jelly of the Umbilical Cord (UC). Similarly to bone marrow MSCs, they have shown multilineage differentiation potential and to be able to provide trophic support to neighboring cells. According to the literature, there are two main populations of cells with a mesenchymal character within the human UC: Wharton's jelly Mesenchymal Stem Cells (WJ-MSCs) and Human Umbilical Cord Perivascular Cells (HUCPVCs). In the present work our aim is to make a comprehensive review on MSC populations of the UC and how these cell populations may be used for future applications in CNS regenerative medicine. Following a brief insight on the general characteristics of MSC like cells, we will discuss the possible sources of stem cells within the WJ and the cord itself (apart UC blood), as well as their phenotypic character. As it has already been shown that these cells hold a strong trophic support to neighbouring cell populations, we will then focus on their secretome, namely which molecules have already been identified within it and their role in phenomena such as immunomodulation. The possible applications of these cell populations to CNS regenerative medicine will be addressed by critically reviewing the work that has been performed so far in this field. Finally, a brief insight will be made on what in the authorsâ opinion are the major challenges in the field for the future application of these cell populations in CNS regenerative medicine.funds attributed by
Fundação Calouste de Gulbenkian to A.J. Salgado under the scope of the The Gulbenkian Programme to Support research in Life Sciences; Portuguese Foundation for Science and Technology (FCT) (PhD scholarship to M.M. Carvalho - SFRH / BD / 51061 / 2010)
Chitosan scaffolds incorporating lysozyme into CaP coatings produced by a biomimetic route : a novel concept for tissue engineering combining a self-regulated degradation system with in situ pore formation
This study describes an innovative self-regulated degrading material with gradual in situ pore formation ability for bone tissue engineering applications. This approach is based on the incorporation of the lysozyme enzyme into calcium phosphate (CaP) coatings, prepared on the surface of chitosan scaffolds by means of a biomimetic coating technique with the aim of controlling their degradation rate and subsequent formation of pores. However, because lysozyme has antibacterial properties, these coatings may act as a carrier for its sustained release, preventing infection upon implantation. In order to prove the concept of in situ pore formation, the coated scaffolds (with and without lysozyme) were incubated in two different solutions at different pH to simulate normal physiological conditions (pH 7.4) and inflammatory response (pH 5). The weight loss and morphology of the scaffolds was monitored over time. At pH 7.4, the scaffolds remained more stable than at pH 5. The scaffolds incubated at pH 5 showed a rapid decrease in their initial weight, and scanning electron microscopy imaging revealed the formation of a highly porous structure. Furthermore, evaluation of the activity of the incorporated lysozyme revealed that the enzyme was able to hydrolyse the peptidoglycan of the bacteria cell walls (as detected by the decrease in optical density with time), indicating that the enzyme remained active after being incorporated into the CaP coating.This work was supported by the European NoE EXPERTISSUES (NMP3-CT-2004-500283), the European STREP HIPPOCRATES (NMP3-CT-2003-505758), the Portuguese Foundation for Science and Technology (FCT) through POCTI and/or FEDER programmes
Fabrication of anisotropically aligned nanofibrous scaffolds based on natural/synthetic polymer blends reinforced with cellulose nanocrystals for tendon tissue engineering
[Excerpt] Introduction: Tendon disorders and injuries are among the most common musculoskeletal problems and their regeneration after injury remains a significant challenge. Given the prevalent fibrous nature of tendonsâ ECM, which exhibits an aligned and hierarchical organization in structures from the nano to the macro scale, uniaxial aligned electrospun nanofibers produced from natural/synthetic polymer blends are among the most successful tendon scaffolds in tissue engineering (TE) strategies[1]. These biomaterials can provide the topographical cues to direct cell adhesion and proliferation, as well as positively affecting cellâs differentiation, phenotype maintenance and matrix deposition. However, the limited mechanical properties of electrospun biomaterials restrict their potential application in this field. In the present study, we propose the use of cellulose nanocrystals (CNCs), the ânature carbon nanotubesâ, as a strategy for the reinforcement of electrospun polyÂÉÂcaprolactoneÂchitosan (PCLÂC) nanofiber scaffolds without compromising their biological performance and thus expand their potential range of application in tendon TE strategies. (...
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