371 research outputs found

    In Vitro Bone Formation Associated with Apatite Coated Polylactide

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    Bone formation onto poly(L-lactide), which was plasma-spray coated with various quantities of hydroxyapatite (0%, 15%, 36% and 100% coverage), was investigated in an in vitro assay. Rat bone marrow cells were grown on the different coatings and the cellular response and elaborated extracellular matrix was examined at the light and electron microscopical level after 1, 2 , 4 and 8 weeks of culture. Proliferation of cells into multilayers was seen on the 0% , 36% and 100% , but not on the 15 % coatings. Coinciding with this was the sparse formation of extracellular matrix on the latter, and its abundant appearance on the former three coatings. Scanning and transmission electron microscopy revealed a mineralized extracellular matrix on the 100% and 36% coatings after 2 and 4 weeks , respectively, and on the 15 % coating after 8 weeks. Mineralization was not observed on uncoated poly(L-lactide). At the interface between hydroxyapatite and the mineralized extracellular matrix, one or more electron dense layers were frequently observed , which showed morphological similarities with structures between these two entities in vivo. The results of this in vitro study show that, in the model used, hydroxyapatite is required to obtain the elaboration of mineralized extracellular matrix on poly(L-lactide)

    Influence of PCL molecular weight on mesenchymal stromal cell differentiation

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    Regenerating or replacing bone, chondral and osteochondral defects, is an active field in tissue engineering. A general strategy is to use a temporary scaffold in which cells are seeded onto the scaffold prior to implantation or attracted into the scaffold from surrounding tissues in the implantation site to form the desired tissue. Several biomaterials have been used for the fabrication of scaffolds, including polycaprolactone (PCL) which is often used for musculoskeletal tissue engineering. The effect of the PCL scaffold architecture on the cell behavior has been investigated. However, the mechanical properties of the bulk material were not taken into account in these studies. PCL is available in a range of molecular weights, resulting in a range of bulk mechanical properties. Since bulk material stiffness is able to direct cell differentiation, it is likely that the molecular weight of PCL may influence cell behavior. Here, we investigated the bulk material properties of both low and high molecular weight PCL scaffolds fabricated through additive manufacturing. The low molecular weight PCL showed a lower bulk material stiffness. During in vitro cell culture, this resulted in a stronger tendency for hypertrophic chondrogenic differentiation compared to the high molecular weight PCL. This study shows that apart from the polymer chemistry and scaffold architecture, the bulk mechanical properties of the polymer used is an important parameter in scaffold fabrication. This is an important finding for the optimization of osteochondral tissue engineering

    Triphasic scaffolds for the regeneration of the bone-ligament interface

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    A triphasic scaffold (TPS) for the regeneration of the bone-ligament interface was fabricated combining a 3D fiber deposited polycaprolactone structure and a polylactic co-glycolic acid electrospun. The scaffold presented a gradient of physical and mechanical properties which elicited different biological responses from human mesenchymal stem cells. Biological test were performed on the whole TPS and on scaffolds comprised of each single part of the TPS, considered as the controls. The TPS showed an increase of the metabolic activity with culturing time that seemed to be an average of the controls at each time point. The importance of differentiation media for bone and ligament regeneration was further investigated. Metabolic activity analysis on the different areas of the TPS showed a similar trend after 7 days in both differentiation media. Total alkaline phosphatase (ALP) activity analysis showed a statistically higher activity of the TPS in mineralization medium compared to the controls. A different glycosaminoglycans amount between the TPS and its controls was detected, displaying a similar trend with respect to ALP activity. Results clearly indicated that the integration of electrospinning and additive manufacturing represents a promising approach for the fabrication of scaffolds for the regeneration of tissue interfaces, such as the bone-to-ligament one, because it allows mimicking the structural environment combining different biomaterials at different scales

    Hydrogels that listen to cells:a review of cell-responsive strategies in biomaterial design for tissue regeneration

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    The past decade has seen a decided move from static and passive biomaterials to biodegradable, dynamic, and stimuli responsive materials in the laboratory and the clinic. Recent advances towards the rational design of synthetic cell-responsive hydrogels-biomaterials that respond locally to cells or tissues without the input of an artificial stimulus-have provided new strategies and insights on the use of artificial environments for tissue engineering and regenerative medicine. These materials can often approximate responsive functions of a cell's complex natural extracellular environment, and must respond to the small and specific stimuli provided within the vicinity of a cell or tissue. In the current literature, there are three main cell-based stimuli that can be harnessed to create responsive hydrogels: (1) enzymes (2) mechanical force and (3) metabolites/small molecules. Degradable bonds, dynamic covalent bonds, and non-covalent or supramolecular interactions are used to provide responsive architectures that enable features ranging from cell selective infiltration to control of stem-cell differentiation. The growing ability to spatiotemporally control the behavior of cells and tissue with rationally designed responsive materials has the ability to allow control and autonomy to future generations of materials for tissue regeneration, in addition to providing understanding and mimicry of the dynamic and complex cellular niche

    3D high throughput screening and profiling of embryoid bodies in thermoformed microwell plates

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    3D organoids using stem cells to study development and disease are now widespread. These models are powerful to mimic in vivo situations but are currently associated with high variability and low throughput. For biomedical research, platforms are thus necessary to increase reproducibility and allow high-throughput screens (HTS). Here, we introduce a microwell platform, integrated in standard culture plates, for functional HTS. Using micro-thermoforming, we form round-bottom microwell arrays from optically clear cyclic olefin polymer films, and assemble them with bottom-less 96-well plates. We show that embryonic stem cells aggregate faster and more reproducibly (centricity, circularity) as compared to a state-of-the-art microwell array. We then run a screen of a chemical library to direct differentiation into primitive endoderm (PrE) and, using on-chip high content imaging (HCI), we identify molecules, including regulators of the cAMP pathway, regulating tissue size, morphology and PrE gene activity. We propose that this platform will benefit to the systematic study of organogenesis in vitro
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