Myoconductive and osteoinductive free-standing polysaccharide membranes

Free-standing (FS) membranes have increasing applications in the biomedical field as drug delivery systems for wound healing and tissue engineering. Here, we studied the potential of free-standing membranes made by the layer-by-layer assembly of chitosan and alginate to be used as a simple biomimetic system of the periosteum. The design of a periosteum-like membrane implies the elaboration of a thick membrane suitable for both muscle and bone formation. Our aim was to produce well-defined ∼50 μm thick polysaccharide membranes that could be easily manipulated, were mechanically resistant, and would enable both myogenesis and osteogenesis in vitro and in vivo. The membranes were chemically crosslinked to improve their mechanical properties. Crosslinking chemistry was followed via Fourier transform infrared spectroscopy and the mechanical properties of the membranes were assessed using dynamic mechanical analysis. The loading and release of the potent osteoinductive growth factor bone morphogenetic protein 2 (BMP-2) inside and outside of the FS membrane was followed by fluorescence spectroscopy in a physiological buffer over 1 month. The myogenic and osteogenic potentials of the membranes in vitro were assessed using BMP-2-responsive skeletal myoblasts. Finally, their osteoinductive properties in vivo were studied in a preliminary experiment using a mouse ectopic model. Our results showed that the more crosslinked FS membranes enabled a more efficient myoblast differentiation in myotubes. In addition, we showed that a tunable amount of BMP-2 can be loaded into and subsequently released from the membranes, depending on the crosslinking degree and the initial BMP-2 concentration in solution. Only the more crosslinked membranes were found to be osteoinductive in vivo. These polysaccharide-based membranes have strong potential as a periosteum-mimetic scaffold for bone tissue regeneration.This work was financially supported by the Foundation for Science and Technology (FCT) through the scholarship SFRH/BPD/96797/2013, Fundo Social Europeu (FSE), and Programa Diferencial de Potencial Human (POPH) granted to Sofia G. Caridade. C.M. is indebted to the Association Francaise contre les Myopathies for financial support via a post-doctoral fellowship (AFM project 16673). J.A. acknowledges the Whitaker International Fellows and Scholars Program for support via a post-doctoral fellowship. This work was supported by the European Commission (FP7 program) via a European Research Council starting grant (BIOMIM, GA 259370 to C.P.) and by the AFM (grant Microtiss, 16530). We thank Isabelle Paintrand for her technical help with the confocal apparatus

Secondary structure of rhBMP-2 in a protective biopolymeric carrier material

Efficient delivery of growth factors is one of the great challenges of tissue engineering. Polyelectrolyte multilayer films (PEM) made of biopolymers have recently emerged as an interesting carrier for delivering recombinant human bone morphogenetic protein 2 (rhBMP-2 noted here BMP-2) to cells in a matrix-bound manner. We recently showed that PEM made of poly(l-lysine) and hyaluronan (PLL/HA) can retain high and tunable quantities of BMP-2 and can deliver it to cells to induce their differentiation in osteoblasts. Here, we investigate quantitatively by Fourier transform infrared spectroscopy (FTIR) the secondary structure of BMP-2 in solution as well as trapped in a biopolymeric thin film. We reveal that the major structural elements of BMP-2 in solution are intramolecular β-sheets and unordered structures as well as α-helices. Furthermore, we studied the secondary structure of rhBMP-2 trapped in hydrated films and in dry films since drying is an important step for future applications of these bioactive films onto orthopedic biomaterials. We demonstrate that the structural elements were preserved when BMP-2 was trapped in the biopolymeric film in hydrated conditions and, to a lesser extent, in dry state. Importantly, its bioactivity was maintained after drying of the film. Our results appear highly promising for future applications of these films as coatings of biomedical materials, to deliver bioactive proteins while preserving their bioactivity upon storage in dry state.This work was supported by the French Ministry of Research through an ANR-EmergenceBIO grant (ANR-09-EBIO-012-01), by the European Commission (FP7 program) via a European Research Council starting grant (BIOMIM, GA 259370), and by GRAVIT (081012_FIBIOS). C.P. is grafetul to IUF for financial support

Osteoinductive coatings of implantable materials

National audienc

Osteoinductive coatings of implantable materials

National audienc

Tectonometamorphic evolution of the Atbashi high- P units (Kyrgyz CAOB, Tien Shan): Implications for the closure of the Turkestan Ocean and continental subduction-exhumation of the South Kazakh continental margin

The South Tien Shan (STS) belt results from the last collision event in the western Central Asian Orogenic Belt (CAOB). Understanding its formation is of prime importance in the general framework of the CAOB. The Atbashi Range preserves high‐P (HP) rocks along the STS suture, but still, its global metamorphic evolution remains poorly constrained. Several HP units have been identified: (a) a HP tectonic mélange including boudins of mafic eclogites in a sedimentary matrix, (b) a large (>100 km long) high‐P metasedimentary unit (HPMU) and (c) a lower blueschist facies accretionary prism. Raman Spectroscopy on carbonaceous material combined with phengite and chlorite multiequilibria and isochemical phase diagram modelling indicates that the HPMU recorded homogeneous P–T conditions of 23–25 kbar and 560–570°C along the whole unit. 40Ar/39Ar dating on phengite from the HPMU ranges between 328 and 319 Ma at regional scale. These ages are interpreted as (re‐) crystallization ages of phengite during Tmax conditions at a pressure range of 20–25 kbar. Thermobarometry on samples from the HP tectonic mélange provides similar metamorphic peak conditions. Thermobarometry on the blueschist to lower greenschist facies accretionary prism indicates that it underwent P–T conditions of 5–6 kbar and 290–340°C, highlighting a 17–20 kbar pressure gap between the HPMU‐tectonic mélange units and the accretionary prism. Comparison with available geochronological data suggests a very short time span between the prograde path (340 Ma), HP metamorphic peak (330 Ma), the Tmax (328–319 Ma) and the final exhumation of the HPMU (303–295 Ma). Extrusion of the HPMU, accommodated by a basal thrust and an upper detachment, was driven by buoyant forces from 70–75 km up to 60 km depth, which directly followed continental subduction and detachment of the HPMU. At crustal depths, extrusion was controlled by collisional tectonics up to shallow levels. Lithological homogeneity of the HPMU and its continental‐derived character from the North Tien Shan suggest this unit corresponds to the hyper‐extended continental margin of the Kazakh continent, subducted southward below the north continental active margin of the Tarim craton. Integration of the available geological data allows us to propose a general geodynamic scenario for Tien Shan during the Carboniferous with a combination of (a) N‐dipping subduction below the Kazakh margin of Middle Tien Shan until 390–340 Ma and (b) S‐dipping subduction of remaining Turkestan marginal basins between 340 and 320 Ma

Sleep: An Open-Source Python Software for Visualization, Analysis, and Staging of Sleep Data

International audienc

Cyclodextrin/Paclitaxel Complex in Biodegradable Capsules for Breast Cancer Treatment

A novel type of biocompatible hollow capsules that combine severable favorable features as a hydrophobic drug carrier, including host–guest complexation in the shell, the unique biological functions of hyaluronic acid (HA), and transport properties of the multilayer shell, was designed and prepared. These capsules were generated by layer-by-layer (LbL) deposition of HA modified with β-cyclodextrin (CD) molecules and poly­(l-lysine) (PLL) on calcium carbonate particles. Simultaneously, paclitaxel (PTX) was loaded in the LbL wall via host–guest interaction. Under physiological conditions, the incorporated anticancer drug was slowly released, and the capsules remained stable. Because the PTX molecules are selectively complexed by CD in the shell, their release can be triggered by the addition of competitive cyclodextrin molecules in the external medium. By incubating the capsules with breast cancer cells (MDA-MB-231), it was found that the cells bound specifically to the capsules through the CD44 receptor of HA that is overexpressed on their surface. Finally, when breast cancer cells were incubated with the PTX-loaded capsules, their viability was found to strongly decrease. All together, these results highlight the potential for these HA–cyclodextrin-containing capsules in anticancer therapy

Secondary Structure of rhBMP‑2 in a Protective Biopolymeric Carrier Material

Efficient delivery of growth factors is one of the great challenges of tissue engineering. Polyelectrolyte multilayer films (PEM) made of biopolymers have recently emerged as an interesting carrier for delivering recombinant human bone morphogenetic protein 2 (rhBMP-2 noted here BMP-2) to cells in a matrix-bound manner. We recently showed that PEM made of poly­(l-lysine) and hyaluronan (PLL/HA) can retain high and tunable quantities of BMP-2 and can deliver it to cells to induce their differentiation in osteoblasts. Here, we investigate quantitatively by Fourier transform infrared spectroscopy (FTIR) the secondary structure of BMP-2 in solution as well as trapped in a biopolymeric thin film. We reveal that the major structural elements of BMP-2 in solution are intramolecular β-sheets and unordered structures as well as α-helices. Furthermore, we studied the secondary structure of rhBMP-2 trapped in hydrated films and in dry films since drying is an important step for future applications of these bioactive films onto orthopedic biomaterials. We demonstrate that the structural elements were preserved when BMP-2 was trapped in the biopolymeric film in hydrated conditions and, to a lesser extent, in dry state. Importantly, its bioactivity was maintained after drying of the film. Our results appear highly promising for future applications of these films as coatings of biomedical materials, to deliver bioactive proteins while preserving their bioactivity upon storage in dry state

No evidence for TSLP pathway activity in human breast cancer

Thymic stromal lymphopoietin (TSLP) is an epithelial cell-derived cytokine that primes dendritic cells for Th2 induction. It has been implicated in different types of allergic diseases. Recent work suggested that TSLP could play an important role in the tumor microenvironment and influence tumor progression, in particular in breast cancer. In this study we systematically assessed the production of TSLP at the mRNA and protein levels in several human breast cancer cell lines, large-scale public transcriptomics data sets, and primary human breast tumors. We found that TSLP production was marginal, and concerned less than 10% of the tumors, with very low mRNA and protein levels. In most cases TSLP was undetectable and found to be expressed at lower levels in breast cancer as compared to normal breast tissue. Last, we could not detect any functional TSLP receptor (TSLPR) expression neither on hematopoietic cells nor on stromal cells within the primary tumor microenvironment. We conclude that TSLP-TSLPR pathway activity is not significantly detected within human breast cancer. Taken together, these observations do not support TSLP targeting in breast cancer