Human adipose stromal cells (ASC) for the regeneration of injured cartilage display genetic stability after in vitro culture expansion

Mesenchymal stromal cells are emerging as an extremely promising therapeutic agent for tissue regeneration due to their multi-potency, immune-modulation and secretome activities, but safety remains one of the main concerns, particularly when in vitro manipulation, such as cell expansion, is performed before clinical application. Indeed, it is well documented that in vitro expansion reduces replicative potential and some multi-potency and promotes cell senescence. Furthermore, during in vitro aging there is a decrease in DNA synthesis and repair efficiency thus leading to DNA damage accumulation and possibly inducing genomic instability. The European Research Project ADIPOA aims at validating an innovative cell-based therapy where autologous adipose stromal cells (ASCs) are injected in the diseased articulation to activate regeneration of the cartilage. The primary objective of this paper was to assess the safety of cultured ASCs. The maintenance of genetic integrity was evaluated during in vitro culture by karyotype and microsatellite instability analysis. In addition, RT-PCR array-based evaluation of the expression of genes related to DNA damage signaling pathways was performed. Finally, the senescence and replicative potential of cultured cells was evaluated by telomere length and telomerase activity assessment, whereas anchorage-independent clone development was tested in vitro by soft agar growth. We found that cultured ASCs do not show genetic alterations and replicative senescence during the period of observation, nor anchorage-independent growth, supporting an argument for the safety of ASCs for clinical use

Assessment of Natural Resources Use for Sustainable Development - DPSIR Framework for Case Studies in Portsmouth and Thames Gateway, U.K.

This chapter reports on the uses of the DPSIR framework to assess the sustainability of the intertidal environments within the two UK case study areas, Portsmouth and Thames Gateway. It focuses on statutory conservation areas dominated by intertidal habitats. Two are located in Portsmouth (Portsmouth and Langstone Harbours) and four in the Thames Gateway (Benfleet Marshes, South Thames Estuary, Medway Estuary and the Swale in the Thames Gateway). Based on the reduction of a number of pressures and impacts observed in recent decades and the improvement of overall environmental quality, all six SSSIs are considered to be sustainable in the short and medium term. In the future, it is possible that the impacts of climate change, especially sea-level rise, might result in further reduction in the area and/or quality of intertidal habitats. Further integration between conservation and planning objectives (both for urban development and management of flood risk) at local level is needed to support the long-term sustainability of intertidal habitats

Lupus anticoagulant identifies two distinct groups of patients with different antibody patterns

Background: Whether antibodies directed to β2-Glycoprotein I (aβ2GPI) are responsible for LA activity is not well defined. However, in the absence of such antibodies the molecule responsible for LA phenomenon is unknown. Objective: The aim of this study was the biochemical identification of the target antigen epitope of aPL responsible of LA activity in the absence of aβ2GPI antibodies together with the biological and clinical characteristics of these patients in comparison with classical triple positive patients. Patients/methods: A comparison of patients with LA without (LA+/aβ2GPI−) and those with (LA+/aβ2GPI+) associated aβ2GPI antibodies was performed. Size exclusion chromatography and analytical chromatography were used to identify the molecule with LA activity in patients LA+/aβ2GPI-. Results and conclusions: Analytical size-exclusion chromatography revealed a peak of 996Kd with LA activity perfectly overlapping that of IgM anti phosphatidylserine/prothrombin (aPS/PT) antibodies. Similarly, all the 25 LA+/aβ2GPI− patients were positive for aPS/PT antibodies. LA+/aβ2GPI− compared to 33 LA+/aβ2GPI+ patients turned out to be significantly older, with a lower rate of previous thromboembolic events and a weaker LA activity. Search for aPS/PT and aβ2GPI antibodies in patients with LA is useful to identify two subgroups of LA at different risk of thromboembolic event

Microstructure and indentation mechanical properties of YSZ nanostructured coatings obtained by suspension plasma spraying

[EN] A commercial nanosuspension of yttria-stabilised zirconia (YSZ) was successfully deposited on austenitic stainless steel substrate by suspension plasma spraying technique (SPS). A SG-100 torch with internal radial injection was used for the spraying. The pneumatic system transported the feed suspension from the containers to the plasma torch. In order to study the effect of the spraying parameters, a factorial model was used to design the experiments, changing both spraying translation speed and suspension flow rate. The coating microstructure was characterised by FEG-SEM. All coatings displayed a two-zone microstructure formed by nanometre-sized particles surrounded by fully molten areas. Moreover, crystalline phases were determined by XRD and Raman spectroscopy. Mechanical properties were also determined using nanoindentation technique. Nanoindentation tests showed a bimodal distribution of the mechanical properties (hardness and Young's modulus) which is related to the two zones (molten and partially molten) present in the coatings. (C) 2012 Elsevier B.V. All rights reserved.This work has been supported by the Spanish Ministry of Science and Innovation (project MAT2009-14144-C03) and the Research Promotion Plan of the Universitat Jaume I, action 2.1 (ref. E-2011-05) and action 3.1 (ref. PREDOC/2009/10). The authors are grateful to Leszek Łatka for his help in plasma spray experiments.Carpio, P.; Rayón Encinas, E.; Pawlowski, L.; Cattini, A.; Benavente Martínez, R.; Bannier, E.; Salvador Moya, MD.... (2013). Microstructure and indentation mechanical properties of YSZ nanostructured coatings obtained by suspension plasma spraying. Surface and Coatings Technology. 220:237-243. https://doi.org/10.1016/j.surfcoat.2012.09.047S23724322

Gallium incorporation into phosphate based glasses: bulk and thin film properties

The osteogenic ions Ca2+, P5+, Mg2+, and antimicrobial ion Ga3+ were homogenously dispersed into a 1.45 mum thick phosphate glass coating by plasma assisted sputtering onto CP grade titanium. The objective was to deliver therapeutic ions in orthopedic/dental implants such as hip prosthesis or dental screws. The hardness 4.7 GPa and elastic modulus 69.7 GPa, of the coating were comparable to plasma sprayed hydroxyapatite/dental enamel, whilst superseding femoral cortical bone. To investigate the manufacturing challenge of translation from a target to vapour condensed coating, structural/compositional properties of the target (P51MQ) were compared to the coating (P40PVD) and a melt-quenched equivalent (P40MQ). Following condensation from P51MQ to P40PVD, P2O5 content reduced from 48.9 to 40.5 mol%. This depolymerisation and reduction in the P-O-P bridging oxygen content as determined by 31P-NMR, FTIR and Raman spectroscopy techniques was attributed to a decrease in the P2O5 network former and increases in alkali/alkali-earth cations. P40PVD appeared denser (3.47 vs. 2.70 g cm-3) and more polymerised than it’s compositionally equivalent P40MQ, showing that structure/ mechanical properties were affected by manufacturing route