Cellen temmen met slimme klei

‘Vorm bot’. ‘Maak bloedvaten’. ‘Ga hechten’. Celbioloog prof. Clemens van Blitterswijk heeft met zijn Twentse onderzoeksgroep een manier gevonden om menselijke voorlopercellen opdrachten te geven. Vandaag ontvangt hij de Federa-prijs

Nano-apatite/polymer composites: mechanical and physicochemical characteristics

Hydrothermally synthesized acicular nano-apatite (Nap) was used as filler to make composites with a polyethylene glycol/poly(butylene terephthalate) (PEG/PBT) block copolymer (Polyactive™70:30). The Nap had a particle diameter of 9–25 nm and a length of 80–200 nm. The mechanical properties and the physiochemical characteristics of the composites, such as Young's modulus, swelling degree in water and the calcification behaviour, have been determined. It was found that Nap had a strong ability to promote the calcification of composites when incorporated into Polyactive 70:30, while poly(acrylic acid) (PAA) coating of Nap had an adverse effect on the calcification of composites, presumably due to the formation of complexes between PAA and PEG segments. Nap had a prominent stiffening effect for Polyactive 70:30 in the dry state, but had a poor stiffening effect for composites in an aqueous environment due to the hygroscopic nature and/or the formation of aggregates. PAA coating on Nap had almost no additional effect on the mechanical properties of composites either in the dry state or in an aqueous environment. To reinforce the polymer by Nap, achieving a more homogeneous dispersion of Nap in the polymer matrix and surface modifications to render the powders less hygroscopic appear to be necessary

Darcian permeability constant as indicator for shear stresses in regular scaffold systems for tissue engineering

The shear stresses in printed scaffold systems for tissue engineering depend on the flow properties and void volume in the scaffold. In this work, computational fluid dynamics (CFD) is used to simulate flow fields within porous scaffolds used for cell growth. From these models the shear stresses acting on the scaffold fibres are calculated. The results led to the conclusion that the Darcian (k 1) permeability constant is a good predictor for the shear stresses in scaffold systems for tissue engineering. This permeability constant is easy to calculate from the distance between and thickness of the fibres used in a 3D printed scaffold. As a consequence computational effort and specialists for CFD can be circumvented by using this permeability constant to predict the shear stresses. If the permeability constant is below a critical value, cell growth within the specific scaffold design may cause a significant increase in shear stress. Such a design should therefore be avoided when the shear stress experienced by the cells should remain in the same order of magnitud

Cell sources for articular cartilage repair strategies: shifting from mono-cultures to co-cultures

The repair of articular cartilage is challenging due to the sparse native cell population combined with the avascular and aneural nature of the tissue. In recent years cartilage tissue engineering has shown great promise. As with all tissue engineering strategies, the possible therapeutic outcome is intimately linked with the used combination of cells, growth factors and biomaterials. However, the optimal combination has remained a controversial topic and no consensus has been reached. In consequence, much effort has been dedicated to further design, investigate and optimize cartilage repair strategies. Specifically, various research groups have performed intensive investigations attempting to identify the single most optimal cell source for articular cartilage repair strategies. However, recent findings indicate that not the heavily investigated mono cell source, but the less studied combinations of cell sources in co-culture might be more attractive for cartilage repair strategies. This review will give a comprehensive overview on the cell sources that have been investigated for articular cartilage repair strategies. In particular, the advantages and disadvantages of investigated cell sources are comprehensively discussed with emphasis on the potential of co-cultures in which benefits are combined while the disadvantages of single cell sources for cartilage repair are mitigated

Flexible (Polyactive®) versus rigid (hydroxyapatite) dental implants

In a beagle dog study, the peri-implant bone changes around flexible (Polyactive®) and rigid hydroxyapatite (HA) implants were investigated radiographically by quantitative digital subtraction analysis and by assessment of marginal bone height, with the aid of a computerized method. A loss of approximately 1 mm of marginal bone height was observed for both the dense Polyactive and the HA implants, after 6 months of loading. This value appeared to be stable from 12 weeks of loading onward. Along the total length of the implant during the first 6 weeks of loading, both the flexible (dense Polyactive) and the rigid (HA) implants showed a decrease in density. However, after this 6-week period, the bone density around the implants increased, and after 18 weeks the original bone density was reached. The flexible Polyactive implants provoked less decrease in density than the rigid HA implants, although not to a statistically significant level. This finding sustains the hypothesis that flexible implant materials may transfer stresses to the surrounding bone more favorably

Factors of having influence on the rheological properties of Ti6A14V slurry

A highly porous Ti6Al4V could be produced with a porous polymeric sponge and Ti6Al4V slurry. However, the rheological properties of Ti6Al4V slurry appeared to be the key issue in the preparation of porous Ti6Al4V. In this study, factors having influence on the rheological properties of Ti6Al4V slurry were addressed in detail. Ti6Al4V powders, organic thickening agents (binders), dispersants, concentration of powder and pH values were optimised with regard to the rheological properties of Ti6Al4V slurry. The results show that Ti6Al4V powder with a mean diameter of 45 μm and spherical shape is beneficial for the preparation of Ti6Al4V slurry. Meanwhile binders with two ingredients, which decompose at different temperatures, have the advantage to keep the shape after debinding. The optimised procedure, based on the findings, made it possible to produce highly porous Ti6Al4V with reticulate porous structure. Porous Ti6Al4V produced by this way is expected to be a promising biomaterial for tissue engineering scaffolds and orthopaedic implant applications

A dual flow bioreactor with controlled mechanical stimulation for cartilage tissue engineering

In cartilage tissue engineering bioreactors can create a controlled environment to study chondrocyte behavior under mechanical stimulation or produce chondrogenic grafts of clinically relevant size. Here we present a novel bioreactor, which combines mechanical stimulation with a two compartment system through which nutrients can be supplied solely by diffusion from opposite sides of a tissue engineered construct. This design is based on the hypothesis that creating gradients of nutrients, growth factors and growth factor antagonists can aid in the generation of zonal tissue engineered cartilage. Computational modeling predicted that the design facilitates the creation of a biologically relevant glucose gradient. This was confirmed by quantitative glucose measurements in cartilage explants. In this system it is not only possible to create gradients of nutrients, but also of anabolic or catabolic factors. Therefore, the bioreactor design allows control over nutrient supply and mechanical stimulation useful for in vitro generation of cartilage constructs that can be used for the resurfacing of articulated joints or as a model for studying OA disease progression

Combining technologies to create bioactive hybrid scaffolds for bone tissue engineering

Combining technologies to engineer scaffolds that can offer physical and chemical cues to cells is an attractive approach in tissue engineering and regenerative medicine. In this study, we have fabricated polymer-ceramic hybrid scaffolds for bone regeneration by combining rapid prototyping (RP), electrospinning (ESP) and a biomimetic coating method in order to provide mechanical support and a physico-chemical environment mimicking both the organic and inorganic phases of bone extracellular matrix (ECM). Poly(ethylene oxide terephthalate)-poly(buthylene terephthalate) (PEOT/PBT) block copolymer was used to produce three dimensional scaffolds by combining 3D fiber (3DF) deposition, and ESP, and these constructs were then coated with a Ca-P layer in a simulated physiological solution. Scaffold morphology and composition were studied using scanning electron microscopy (SEM) coupled to energy dispersive X-ray analyzer (EDX) and Fourier Tranform Infrared Spectroscopy (FTIR). Bone marrow derived human mesenchymal stromal cells (hMSCs) were cultured on coated and uncoated 3DF and 3DF + ESP scaffolds for up to 21 d in basic and mineralization medium and cell attachment, proliferation, and expression of genes related to osteogenesis were assessed. Cells attached, proliferated and secreted ECM on all the scaffolds. There were no significant differences in metabolic activity among the different groups on days 7 and 21. Coated 3DF scaffolds showed a significantly higher DNA amount in basic medium at 21 d compared with the coated 3DF + ESP scaffolds, whereas in mineralization medium, the presence of coating in 3DF+ESP scaffolds led to a significant decrease in the amount of DNA. An effect of combining different scaffolding technologies and material types on expression of a number of osteogenic markers (cbfa1, BMP-2, OP, OC and ON) was observed, suggesting the potential use of this approach in bone tissue engineerin

A Platform of Porous Biomaterials as 3D Culture Systems for Cancer Biology

Background: The role of stem cells in tissue development and repair is beginning to be unravelled and will open remarkable opportunities to improve current medical treatments. Yet, the cascade of events that enable us to distinguish between abnormal and functional tissue morphogenesis is not well known and the potential involvement of stem cells in cancer initiation or tissue regeneration is still at an embryonic stage. Most biological studies rely on culturing cells onto two-dimensional (2D) substrates, which poorly reflect the three-dimensional (3D) environment that governs the physical, chemical, and biological processes at the heart of tissue development. Here, we introduce a library of 3D culture systems − scaffolds − with enhanced cell-material interactions. Materials and Methods: 3D scaffolds made of biodegradable synthetic polymers were fabricated by either rapid prototyping, eletrospinning and their combination. Mesenchymal stem cells derived from bone marrow were isolated from patients after informed consent, seeded on the scaffolds and cultured for up to 35 days. Cell morphology was observed by scanning electron microscopy. Cell number and metabolic activity were quantified by DNA and alamar blue assays. Differentiation was assessed by gene expression, while extrfacellular matrix (ECM) formation by biochemical assays. Results: 3D scaffolds with tailored mechanical and physichochemical properties could be fabricated by different processing technologies. While rapid prototyping resulted in the fabrication of scaffolds with controlled porosity at the macro scale, electrospinning enabled the creation of fibrillar meshes mimicking the physical micro and nano scale dimensions of native ECM. Nutrient availability had a profound effect on tissue formation in 2D and 3D. Despite steep nutrient concentration gradients in 3D scaffolds, stem cells proliferated while avoiding significant death. Cell migration into millimeter-size circular patterns in the scaffold’s pores was supported by ECM organization. Higher concentrations of nutrients controlled the rates of proliferation and did not induce differentiation markers. Furthermore, scaffolds with customized physicochemical and surface properties influenced stem cell morphology and activity. Conclusions: These 3D scaffolds offer a new platform to study the mechanisms behind stem cell driven tissue morphogenesis and may play a role in cancer biology research to create organotypic 3D models to study cancer initiation and development, as well as the potential involvement of stem cells in these processes

Controlled surface initiated polymerization of N-isopropylacrylamide from polycaprolactone substrates for regulating cell attachment and detachment

Poly(ε-caprolactone) (PCL) substrates were modified with thermoresponsive poly(N-isopropylacrylamide) (PNIPAM) brushes to direct and control cellular attachment and detachment. Prior to brush growth, the surface of PCL was activated by a diamine to allow for initiator coupling. Infrared spectra taken before and after cell culturing demonstrated the covalently attached nature of the PNIPAM brushes. PCL is a biocompatible polymer and to prove that the modifications described above did not change this characteristic property, a cell attachment/detachment study was carried out. The modified substrates showed a lower cell attachment when compared to PCL alone and to PCL films modified with the initiator. The possibility to detach the cells in the form of a sheet was proved using PNIPAM-modified PCL films by lowering the temperature to 25 °C. No relevant detachment was shown by the unmodified or by the initiator modified surfaces. This confirmed that the detachment was temperature dependent and not connected to other factors such as polymer swelling. These functionalized polymeric films can find applications as smart cell culture systems in regenerative medicine applications