Karine Anselme

Université de Haute-Alsace, Mulhousen, Alsace, France

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Publications (148)387.89 Total impact

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    ABSTRACT: Samples of crystalline hydroxyapatite (HA) with and without the addition of individual Mg2 +, Mn2 + and Sr2 + ions and samples with the addition of all three ions simultaneously were prepared using the precipitation method in an aqueous medium. Chemical, structural, spectroscopic and thermophysical analyses of the synthesized samples were conducted. The obtained results indicate that Sr2 + ions were easily incorporated into the HA crystal structure, whereas it was difficult to incorporate Mg2 + and Mn2 + ions into the HA lattice when these ions were individually introduced into the samples. The synthesis of HA with Mg2 + or Mn2 + ions is characterized by the formation of HA with a low concentration of doping elements that is outweighed by the amount of these atoms present in less biocompatible phases that formed simultaneously. However, the incorporation of Sr2 + along with Mg2 + and Mn2 + ions into the samples allowed for the synthesis of HA with considerably higher concentrations of Mg2 + and Mn2 + in the crystal lattice.
    No preview · Article · Apr 2016
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    ABSTRACT: The purpose of this study is to evaluate the potential of simple high performance liquid chromatography (HPLC) setup for quantification of adsorbed proteins on various type of plane substrates with limited area (< 3cm(2)). Protein quantification was investigated with a liquid chromatography chain equipped with a size exclusion column or a reversed-phase column. By evaluating the validation of the method according to guidelines of the International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH), all the results obtained by HPLC were reliable. By simple adsorption test at the contact of hydrophilic (glass) and hydrophobic (polydimethylsiloxane: PDMS) surfaces, kinetics of adsorption were determined and amounts of adsorbed bovine serum albumin, myoglobin and lysozyme were obtained: as expected for each protein, the amount adsorbed at the plateau on glass (between 0.15μg/cm(2) and 0.4μg/cm(2)) is lower than for hydrophobic PDMS surfaces (between 0.45μg/cm(2) and 0.8μg/cm(2)). These results were consistent with bicinchoninic acid protein determination. According to ICH guidelines, both Reversed Phase and Size Exclusion HPLC can be validated for quantification of adsorbed protein. However, we consider the size exclusion approach more interesting in this field because additional informations can be obtained for aggregative proteins. Indeed, monomer, dimer and oligomer of bovine serum albumin (BSA) were observed in the chromatogram. On increasing the temperature, we found a decrease of peak intensity of bovine serum albumin as well as the fraction of dimer and oligomer after contact with PDMS and glass surface. As the surface can act as a denaturation parameter, these informations can have a huge impact on the elucidation of the interfacial behavior of protein and in particular for aggregation processes in pharmaceutical applications.
    Full-text · Article · Jan 2016 · International Journal of Pharmaceutics
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    ABSTRACT: Cancer cells are known to have alterations compared to healthy cells, but can these differences extend to the way cells interact with their environment? Here, the authors focused on the alignment on an array of grooves of nanometer depth using two cell types: healthy osteoprogenitor primary cells (HOP) and a cancerous osteosarcoma (SaOs-2) cell line. Another concern was how this alignment affects the cell&apos;s interior, namely, the nucleus. Based on the results, it is proposed that these two cell types respond to different size regimes: SaOs-2 cells are more sensitive to shallow grooves while HOP cells are strongly aligned with deep grooves. As a measure of the impact of cell alignment on the nucleus the orientation and elongation of the nucleus were determined. Compared to HOP cells, the cell nucleus of SaOs-2 cells is more aligned and elongated in response to grooves, suggesting a softer nucleus and/or increased force transmission. These results support the hypothesis that cancer cells have reduced nucleus rigidity compared to healthy ones and further indicate differences in sensing, which may be important during metastasis.
    Full-text · Article · Aug 2015 · Biointerphases
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    O M Ba · M Hindie · P Marmey · O Gallet · K Anselme · A Ponche · A.C. Duncan
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    ABSTRACT: Quantity, orientation, conformation and covalent linkage of naturally cell adhesive proteins adsorbed or covalently linked to a surface, are known to influence the preservation of their subsequent long term cell adhesion properties and bioactivity. In the present work, we explore two different strategies for the covalent linking of plasma fibronectin (pFN) – used as a cell adhesive model protein, onto a polystyrene (PS) surface. One is aimed at tethering the protein to the surface in a semi-oriented fashion (via one of the 4 free thiol reactive groups on the protein) with a heterofunctional coupling agent (SSMPB method). The other aims to immobilize the protein in a more random fashion by reaction between the abundant pendant primary amine bearing amino acids of the pFN and activated carboxylic surface functions obtained after glutaric anhydride surface treatment (GA method). The overall goal will be to verify the hypothesis of a correlation between covalent immobilization of a model cell adhesive protein to a PS surface in a semi-oriented configuration (versus randomly oriented) with promotion of enhanced exposure of the protein's cell binding domain. This in turn would lead to enhanced cell adhesion. Ideally the goal is to elaborate substrates exhibiting a long term stable protein monolayer with preserved cell adhesive properties and bioactivity for biomaterial and/or cell adhesion commercial plate applications. However, the initial restrictive objective of this paper is to first quantitatively and qualitatively investigate the reversibly (merely adsorbed) versus covalently irreversibly bound protein to the surface after the immobilization procedure.
    Full-text · Article · Jun 2015 · Colloids and surfaces B: Biointerfaces
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    ABSTRACT: The goal of this work was to study the thermal stabilization of calcium apatites in which the Ca2+ ions were substituted for Sr2+ in increasing concentrations via ionic co-substitutions. Two distinct standard syntheses were proposed for comparative purposes: one using counter-ions that were not easily incorporated into the apatite structure (NH4+/NO3−) and one using counter-ions that can be easily incorporated into the structure (Na+/Cl−). After calcination, only the apatites synthesized in the presence of NH4+/NO3− presented phase transformation. In contrast, the apatites synthesized in the presence of Na+/Cl− formed a solid solution after calcination, with Na+, Ca2+, Sr2+ and Cl− sharing the same apatite lattice. Wavelength dispersive X-ray fluorescence spectroscopy (WDXRF), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and temperature-programmed desorption (TPD) techniques showed that the counter-ions present during the syntheses that are associated with CO32− play an important role in the thermal stabilization of the apatites.
    No preview · Article · May 2015 · Materials Science and Engineering B
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    ABSTRACT: To discriminate the most important physicochemical parameters for bone reconstruction, the inflammatory potential of seven nanoporous hydroxyapatite powders synthesized by hard or soft templating was evaluated both in vitro and in vivo. After physical and chemical characterization of the powders, we studied the production of inflammatory mediators by human primary monocytes after 4 and 24 h in contact with powders, and the host response after 2 weeks implantation in a mouse critical size defect model. In vitro results highlighted increases in the secretion of TNF-α, IL-1, -8, -10 and proMMP-2 and -9 and decreases in the secretion of IL-6 only for powders prepared by hard templating. In vivo observations confirmed an extensive inflammatory tissue reaction and a strong resorption for the most inflammatory powder in vitro. These findings highlight that the most critical physicochemical parameters for these nanoporous hydroxyapatite are, the crystallinity that controls dissolution potential, the specific surface area and the size and shape of crystallites.
    Full-text · Article · Mar 2015 · Nanomedicine
  • Melanie Eichhorn · Cleo Stannard · Karine Anselme · Jürgen Rühe
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    ABSTRACT: It has been previously shown that osteosarcoma (SaOs-2) cells respond to micropillared surfaces consisting of poly-l-lactic acid with strong deformation of the cell body and nucleus. Until now, cell nucleus deformation of SaOs-2 cells was only studied by exposing them to square shaped micropillars in an isotropic pattern. Here we report on experiments of the cell nucleus response of such cells to rhombic structures of different topographies generated from a rubbery polymer, namely poly(n-butyacrylate). It is observed that cells orientate themselves perpendicular to the long axis of the rhombi. While their spreading on the surface is not influenced by the opening angle of the structures, rhombic structures with sharper angles induce stronger deformation of the cells and accordingly more elongated nuclei. Graphical abstract
    No preview · Article · Feb 2015 · Journal of Materials Science Materials in Medicine
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    ABSTRACT: The lifetime of bone implants inside the human body is directly related to their osseointegration. Ideally, future materials should be inspired by human tissues and provide the material structure-function relationship from which synthetic advanced biomimetic materials capable of replacing, repairing or regenerating human tissues can be produced. This work describes the development of biomimetic thin coatings on titanium implants to improve implant osseointegration. The assembly of an inorganic-organic biomimetic structure by UV laser pulses is reported. The structure consists of a nanostructured hydroxyapatite (HA) film grown onto a titanium substrate by pulsed laser deposition (PLD) and activated by a top fibronectin (FN) coating deposited by matrix assisted pulsed laser evaporation (MAPLE). A pulsed KrF* laser source (λ=248 nm, τ=25 ns) was employed at fluences of 7 and 0.7J/cm2 for HA and FN transfer, respectively. Films approximately 1500 and 450 nm thick were obtained for HA and FN, respectively. A new cryogenic temperature-programmed desorption mass spectrometry analysis method was employed to accurately measure the quantity of immobilized protein. We determined that less than 7µg FN per cm2 HA surface is adequate to improve adhesion, spreading, and differentiation of osteoprogenitor cells. We believe that the proposed fabrication method opens the door to combining and immobilizing two or more inorganic and organic materials on a solid substrate in a well-defined manner. The flexibility of this method enables the synthesis of new hybrid materials by simply tailoring the irradiation conditions according to the thermo-physical properties of the starting materials.
    No preview · Article · Dec 2014 · ACS Applied Materials & Interfaces

  • No preview · Conference Paper · Nov 2014
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    ABSTRACT: Directional wettability, i.e. the variation of wetting properties, depending on the surface orientation, can be achieved by anisotropic surface texturing. A new high-precision process can produce homogeneous sinusoidal surfaces (in particular, parallel grooves) at the microscale, with a nanoscale residual roughness five orders of magnitude smaller than the texture features. Static wetting experiments have shown that this pattern, even with a very small aspect ratio, can induce a strong variation of the contact angle, depending on the direction of the observation. A comparison with numerical simulations (using Surface Evolver software) shows good agreement and could be used to predict fluid–solid interaction and droplet behaviour on textured surfaces. Two primary mechanisms of directional spreading of water droplets on textured stainless steel surface have been identified. The first one is the mechanical barrier created by the textured surface peaks; this limits spreading in a perpendicular direction to the surface anisotropy. The second one is the capillary action inside of the sinusoidal grooves, which accelerates spreading along the grooves. Spreading has been shown to depend strongly on the history of wetting and internal drop dynamics.
    Full-text · Article · Nov 2014

  • No preview · Conference Paper · Nov 2014
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    ABSTRACT: The objective of this study was to synthesize and characterize novel nano-silver poly(acrylic acid) (AgNP-PAA) via photoreduction technique, and evaluate the biocompatibility and mechanical strength of the formed glass–ionomer cements (GIC). The polymer in the cement was composed of a long-chain poly(acrylic acid) (PAA) solution and AgNO3 (0, 0.05, 0.10 and 0.50 % wt/wt), and tartaric acid (TA) were added to formulate different final concentrations of silver nanoparticles polyacids. The formulations were UV irradiated and UV-Vis spectroscopy monitored the formation of AgNP. Human gingival fibroblasts HGF-1 (ATCC® CRL-2014™) were cultivated using the 24 h DMEM extract of the GIC. The cements were evaluated by compressive strength (CS) and in vitro indirect cytotoxicity test using an MTT assay. In the groups containing high AgNP levels, CS increased by 32 % compared to the control GIC group. Moreover, the synthesis of AgNP in the polyacids of the GIC increased the biocompatibility of the cements by 52 % compared to the control group. The PAA used in this work are not normally used to pharmaceutical needs. Therefore, we hypothesized that the process of photoredution used to synthesize AgNP in the polyacids helped to increase the biocompatibility of this PAA.
    Full-text · Conference Paper · Aug 2014
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    ABSTRACT: Defined protein quantities were embedded in situ in a biodegradable polymer coating during simultaneous laser vaporization of two targets. Fibronectin (FN) and poly-DL-lactide (PDLLA) were transferred and immobilized concomitantly by Combinatorial Matrix Assisted Pulsed Laser Evaporation onto solid substrates. The film surface with gradient of composition was characterized by optical, scanning electron microscopy and profilometry. Micrometric FN packages were visualized in the polymeric matrix by confocal microscopy. The composition of FN was investigated by FUR and tiFTIR analyses in a polymeric matrix with different thickness.
    No preview · Article · Jul 2014 · Applied Surface Science
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    Sylvain Giljean · Maxence Bigerelle · Karine Anselme
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    ABSTRACT: In this study, two series of 11 samples of TiAl6V4 titanium alloy and 316L stainless steel have been polished in an isotropic manner at different levels in order to quantify the influence of biomaterial roughness on cell behavior. Topography was measured by a tactile profilometer and a multiscale analysis has been carried out. Human osteoblasts have been cultured on those samples. It appears that roughness has no reproducible effect on the cell behavior except an influence on cell orientation on the wider grooves. As a conclusion, biomaterial surface damage, in the roughness range between R(a) = 0.01 and 0.1 μm, has no influence on cell-adhesion mechanisms when roughness is isotropic and groove width is inferior to a critical value. SCANNING 00: 1-9, 2012. © 2012 Wiley Periodicals, Inc.
    Full-text · Article · Feb 2014 · Scanning
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    K Anselme · M Bigerelle
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    ABSTRACT: Surface characteristics of materials, whether their topography, chemistry, or surface energy, play an essential part in osteoblast adhesion on biomaterials. Thus, the quality of cell adhesion will influence the cell's capacity to proliferate and differentiate in contact with a biomaterial. We have developed for more than ten years numerous studies on the influence of topography and chemistry of metallic substrates on the response of primary human bone cells. The originality of our approach is that contrary to most of other authors, we quantified the adhesion of primary human bone cells on metallic substrates with perfectly characterized surface topography after some hours but also over 21 days. Moreover, we have developed original statistical approaches for characterizing the relation between surface roughness and cell-adhesion parameters. In this article, we will illustrate different studies we did these last ten years concerning the development of a new adhesion parameter, the adhesion power; the correlation between short-term adhesion, long-term adhesion, and proliferation; the influence of roughness organization on cell adhesion and the development of the order parameter; our modeling approach of cell adhesion on surface topography; the relative influence of surface chemistry and topography on cell adhesion and contact angle; the relation between surface features dimensions and cell adhesion. Further, some considerations will be given on the methods for scanning surface topography for cell-adhesion studies. Finally, perspectives will be given to elucidate these intracellular mechanotransduction mechanisms induced by the deformation of cells on model sinusoidal peaks-or-valleys surfaces. SCANNING 00: 1-10, 2012. © 2012 Wiley Periodicals, Inc.
    Preview · Article · Feb 2014 · Scanning
  • Judith Böhmler · Arnaud Ponche · Karine Anselme · Lydie Ploux
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    ABSTRACT: Highly controlled mixed molecular layers are crucial to study the role of material surface chemistry in biointerfaces, such as bacteria and subsequent biofilms interacting with biomaterials. Silanes with non-nucleophilic functional groups are promising to form self-assembled monolayers (SAMs) due to their low sensitivity to side-reactions. Nevertheless, the real control of surface chemistry, layer structure and organization has not been determined. Here, we report a comprehensive synthesis and analysis of undecyltrichlorosilane- and 11-bromoundecyltrichlorosilane-based mixed SAMs on silicon substrates. The impact of the experimental conditions on the control of surface chemistry, layer structure and organization was investigated by combining survey and high-resolution X-ray photoelectron spectroscopy analysis, wettability measurements and ellipsometry. The most appropriate conditions were first determined for elaborating highly reproducible, but easily made, pure 11-bromoundecyltrichlorosilane SAMs. We have demonstrated that the control is maintained on more complex surfaces i.e., surfaces revealing various chemical densities, which were obtained with different ratios of undecyltrichlorosilane and 11-bromoundecyltrichlorosilane. The control is also maintained after bromine to amine group conversion via SN2 bromine-to-azide reactions. The appropriateness of such highly controlled amino- and methyl-group revealing platforms (NH2-X%/CH3) for biointerface studies was shown by the higher reproducibility of bacterial adhesion on NH2-100%/CH3 SAMs compared to bacterial adhesion on molecular layers of overall similar surface chemistry, but less control at the molecular scale.
    No preview · Article · Oct 2013 · ACS Applied Materials & Interfaces
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    ABSTRACT: Arrays of nanowires (NWs) are currently being established as vehicles for molecule delivery and electrical- and fluorescence-based platforms in the development of biosensors. It is conceivable that NW-based biosensors can be optimized through increased understanding of how the nanotopography influences the interfaced biological material. Using state-of-the-art homogenous NW arrays allows for a systematic investigation of how the broad range of NW densities used by the community influences cells. Here it is demonstrated that indium arsenide NW arrays provide a cell-promoting surface, which induces both increased cell proliferation and focal adhesion up-regulation. Furthermore, a systematic variation in NW spacing affects both the detailed cell morphology and adhesion properties, where the latter can be predicted based on changes in free-energy states using the proposed theoretical model. As the NW density influences cellular parameters, such as cell size and adhesion tightness, it will be important to take NW density into consideration in the continued development of NW-based platforms for cellular applications, such as molecule delivery and electrical measurements.
    No preview · Article · Sep 2013 · ACS Applied Materials & Interfaces
  • L. PAIVA · L.P. DA COSTA · L. PLOUX · R.M. THIR · K. ANSELME
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    ABSTRACT: Objective: The aim of this study was to use a novel synthesized silver nanoparticle (NAg)-containing polyacid to formulate conventional glass ionomer (GI) and evaluate the effect of NAg on the compressive strength and contact inhibition of Streptococcus mutans to the surface of the formed GI. Method: Poly(acrylic acid) (PAA) solutions containing or not NAg synthesized in situ were hand mixed to glass powders of Fuji IXTM (GC coorp.). Compressive strength of groups (n=8) were tested according to ISO 9917:2003. Contact inhibition of S. mutans (CIP 103220) incubated in BHI for 20h at 37oC were compared on GI surfaces using a confocal laser scanning microscopy. Result: The presence of 0.07% NAg in the glass ionomer matrix increased values of compressive strength compared to control (no NAg) (P<0.01). Fluorescent phase-contrast images suggest that NAg are inhibiting S. mutans adhesion due to the reduced amount of bacteria adhered to the surface of NAg groups, and also because of the tendency of agglomeration and changes in cell morphology. Conclusion: This study developed a novel antibacterial conventional glass-ionomer. Within the limitations of this study, it appears that the experimental cement is a clinically attractive dental restorative due to its high mechanical strength and antibacterial function.
    No preview · Conference Paper · Sep 2013
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    ABSTRACT: This work describes the preparation and characterization of porous 3D-scaffolds based on chitosan (CHI), chitosan/silk fibroin (CHI/SF) and chitosan/silk fibroin/hydroxyapatite (CHI/SF/HA) by freeze drying. The biomaterials were characterized by X-ray diffraction, attenuated total reflection Fourier transform infrared spectroscopy, thermogravimetric analysis, differential scanning calorimetry, scanning electron microscopy and energy dispersive spectroscopy. In addition, studies of porosity, pore size, contact angle and biological response of SaOs-2osteoblastic cells were performed. The CHI scaffolds have a porosity of 94.2±0.9%, which is statistically higher than the one presented by CHI/SF/HA scaffolds, 89.7±2.6%. Although all scaffolds were able to promote adhesion, growth and maintenance of osteogenic differentiation of SaOs-2 cells, the new 3D-scaffold based on CHI/SF/HA showed a significantly higher cell growth at 7days and 21days and the level of alkaline phosphatase at 14 and 21days was statistically superior compared to other tested materials.
    No preview · Article · Aug 2013
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    ABSTRACT: Introduction Experimental Methods The Interaction of Bone Cells with Micropillars The Deformation of Skin Cells as a Function of Their Malignancy The Deformation of Fibroblasts of Different Cellular Ages Discussion Conclusions Acknowledgments References
    No preview · Chapter · Jul 2013

Publication Stats

5k Citations
387.89 Total Impact Points

Institutions

  • 2009-2015
    • Université de Haute-Alsace
      • Institut de Science des Matériaux de Mulhouse : IS2M
      Mulhousen, Alsace, France
  • 2008-2015
    • French National Centre for Scientific Research
      Lutetia Parisorum, Île-de-France, France
  • 2014
    • Curtin University Sarawak
      • School of Engineering and Science
      Miri, Sarawak, Malaysia
  • 2009-2014
    • Institut de Science des Matériaux de Mulhouse
      Mulhousen, Alsace, France
  • 2010
    • University of South Australia
      Tarndarnya, South Australia, Australia
  • 2000-2006
    • Université du Littoral Côte d'Opale (ULCO)
      Dunkirk, Nord-Pas-de-Calais, France
  • 2002
    • Institut Calot
      Berck-sur-Mer, Nord-Pas-de-Calais, France