Biocompatibility of poly(lactic acid) with incorporated graphene-based materials

The incorporation of graphene-based materials has been shown to improve mechanical properties of poly(lactic acid) (PLA). In this work, PLA films and composite PLA films incorporating two graphene-based materials – graphene oxide (GO) and graphene nanoplatelets (GNP) – were prepared and characterized regarding not only biocompatibility, but also surface topography, chemistry and wettability. The presence of both fillers changed the films surface topography, increasing the roughness, and modified the wettability – the polar component of surface free energy increased 59% with GO and decreased 56% with GNP. Mouse embryo fibroblasts incubated with both fillers exceeded the IC50 in both cases with a concentration of 10 μg mL−1. No variations in cell proliferation at the surface of the composite films were observed, except for those containing GO after 24 h incubation, which presented higher cell proliferation than pristine PLA films. Platelet adhesion to PLA and PLA/GNP films was lower in the presence of plasma proteins than when no proteins were present. Furthermore, incorporation of GNP into PLA reduced platelet activation in the presence of plasma proteins. The results indicated that low concentrations of GO and GNP may be incorporated safely in PLA to improve aspects relevant for biomedical applications, such as mechanical properties.Funding for this work was partially provided by FEDER, through Programa Operacional Factores de Competitividade - COMPETE, and by National Funding through FCT - Fundacao para a Ciencia e a Tecnologia, in the framework of project PTDC/EME-PME/114808/2009 and of grant SFRH/BPD/63722/2009

Sodium affects the sperm motility in the European eel

[EN] The role of seminal plasma sodium and activation media sodium on sperm motility was examined by selectively removing the element from these two media, in European eel sperm. Sperm size (sperm head area) was also measured using an ASMA (Automated Sperm Morphometry Analyses) system, in the different conditions. Intracellular sodium [Na+](i) was quantitatively analyzed by first time in the spermatozoa from a marine fish species. Measurement of [Na+](i) was done before and after motility activation, by Flow Cytometry, using CoroNa Green AM as a dye. Sperm motility activation induced an increase in [Na+](i) from 96.72 mM in quiescent stage to 152.21 mM post-activation in seawater. A significant decrease in sperm head area was observed post activation in seawater. There was a notable reduction in sperm motility when sodium was removed from the seminal plasma, but not when it was removed from the activation media. Sodium removal was also linked to a significant reduction in sperm head area in comparison to the controls. Our results indicate that the presence of the ion Na+ in the seminal plasma (or in the extender medium) is necessary for the preservation of sperm motility in European eel, probably because it plays a role in maintaining an appropriate sperm cell volume in the quiescent stage of the spermatozoa. (C) 2016 Elsevier Inc All rights reserved.Funded from the SPERMOT project (Spanish Ministry of Science and Innovation, MICINN; AGL2010-16009). M.C. Vilchez has a predoctoral grant from UPV PAID Program (2011-S2-02-6521), Marina Morini has a predoctoral grant from Generalitat Valenciana (Programa Grisolia, GRISOLIA/2012/006), Victor Gallego has a postdoctoral contract from UPV (PAID-10-14), and David S. Penaranda was supported by MICINN (PTA2011-4948-I) and UPV (PTA2011-4948-I). Grants to attend meetings were received from COST Office (Food and Agriculture COST Action FA1205: AQUAGAMETE).Vilchez Olivencia, MC.; Morini, M.; Peñaranda, D.; Gallego Albiach, V.; Asturiano Nemesio, JF.; Pérez Igualada, LM. (2016). Sodium affects the sperm motility in the European eel. Comparative Biochemistry and Physiology - B: Comparative Biochemistry. 198:51-58. https://doi.org/10.1016/j.cbpa.2016.04.008S515819

Guided assembly of nanoparticles on electrostatically charged nanocrystalline diamond thin films

We apply atomic force microscope for local electrostatic charging of oxygen-terminated nanocrystalline diamond (NCD) thin films deposited on silicon, to induce electrostatically driven self-assembly of colloidal alumina nanoparticles into micro-patterns. Considering possible capacitive, sp2 phase and spatial uniformity factors to charging, we employ films with sub-100 nm thickness and about 60% relative sp2 phase content, probe the spatial material uniformity by Raman and electron microscopy, and repeat experiments at various positions. We demonstrate that electrostatic potential contrast on the NCD films varies between 0.1 and 1.2 V and that the contrast of more than ±1 V (as detected by Kelvin force microscopy) is able to induce self-assembly of the nanoparticles via coulombic and polarization forces. This opens prospects for applications of diamond and its unique set of properties in self-assembly of nano-devices and nano-systems

Modified Vaccinia Virus Ankara-Based Vaccine Vectors Induce Apoptosis in Dendritic Cells Draining from the Skin via both the Extrinsic and Intrinsic Caspase Pathways, Preventing Efficient Antigen Presentation

Dendritic cells (DC) are potent antigen-presenting cells and central to the induction of immune responses following infection or vaccination. The collection of DC migrating from peripheral tissues by cannulation of the afferent lymphatic vessels provides DC which can be used directly ex vivo without extensive in vitro manipulations. We have previously used bovine migrating DC to show that recombinant human adenovirus 5 vectors efficiently transduce afferent lymph migrating DEC-205(+) CD11c(+) CD8(-) DC (ALDC). We have also shown that recombinant modified vaccinia virus Ankara (MVA) infects ALDC in vitro, causing downregulation of costimulatory molecules, apoptosis, and cell death. We now show that in the bovine system, modified vaccinia virus Ankara-induced apoptosis in DC draining from the skin occurs soon after virus binding via the caspase 8 pathway and is not associated with viral gene expression. We also show that after virus entry, the caspase 9 pathway cascade is initiated. The magnitude of T cell responses to mycobacterial antigen 85A (Ag85A) expressed by recombinant MVA-infected ALDC is increased by blocking caspase-induced apoptosis. Apoptotic bodies generated by recombinant MVA (rMVA)-Ag85A-infected ALDC and containing Ag85A were phagocytosed by noninfected migrating ALDC expressing SIRPα via actin-dependent phagocytosis, and these ALDC in turn presented antigen. However, the addition of fresh ALDC to MVA-infected cultures did not improve on the magnitude of the T cell responses; in contrast, these noninfected DC showed downregulation of major histocompatibility complex class II (MHC-II), CD40, CD80, and CD86. We also observed that MVA-infected ALDC promoted migration of DEC-205(+) SIRPα(+) CD21(+) DC as well as CD4(+) and CD8(+) T cells independently of caspase activation. These in vitro studies show that induction of apoptosis in DC by MVA vectors is detrimental to the subsequent induction of T cell responses

Enhanced Growth and Osteogenic Differentiation of Human Osteoblast-Like Cells on Boron-Doped Nanocrystalline Diamond Thin Films

Intrinsic nanocrystalline diamond (NCD) films have been proven to be promising substrates for the adhesion, growth and osteogenic differentiation of bone-derived cells. To understand the role of various degrees of doping (semiconducting to metallic-like), the NCD films were deposited on silicon substrates by a microwave plasma-enhanced CVD process and their boron doping was achieved by adding trimethylboron to the CH4:H2 gas mixture, the B∶C ratio was 133, 1000 and 6700 ppm. The room temperature electrical resistivity of the films decreased from >10 MΩ (undoped films) to 55 kΩ, 0.6 kΩ, and 0.3 kΩ (doped films with 133, 1000 and 6700 ppm of B, respectively). The increase in the number of human osteoblast-like MG 63 cells in 7-day-old cultures on NCD films was most apparent on the NCD films doped with 133 and 1000 ppm of B (153,000±14,000 and 152,000±10,000 cells/cm2, respectively, compared to 113,000±10,000 cells/cm2 on undoped NCD films). As measured by ELISA per mg of total protein, the cells on NCD with 133 and 1000 ppm of B also contained the highest concentrations of collagen I and alkaline phosphatase, respectively. On the NCD films with 6700 ppm of B, the cells contained the highest concentration of focal adhesion protein vinculin, and the highest amount of collagen I was adsorbed. The concentration of osteocalcin also increased with increasing level of B doping. The cell viability on all tested NCD films was almost 100%. Measurements of the concentration of ICAM-1, i.e. an immunoglobuline adhesion molecule binding inflammatory cells, suggested that the cells on the NCD films did not undergo significant immune activation. Thus, the potential of NCD films for bone tissue regeneration can be further enhanced and tailored by B doping and that B doping up to metallic-like levels is not detrimental for cells

The Advancement of Biomaterials in Regulating Stem Cell Fate.

Stem cells are well-known to have prominent roles in tissue engineering applications. Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) can differentiate into every cell type in the body while adult stem cells such as mesenchymal stem cells (MSCs) can be isolated from various sources. Nevertheless, an utmost limitation in harnessing stem cells for tissue engineering is the supply of cells. The advances in biomaterial technology allows the establishment of ex vivo expansion systems to overcome this bottleneck. The progress of various scaffold fabrication could direct stem cell fate decisions including cell proliferation and differentiation into specific lineages in vitro. Stem cell biology and biomaterial technology promote synergistic effect on stem cell-based regenerative therapies. Therefore, understanding the interaction of stem cell and biomaterials would allow the designation of new biomaterials for future clinical therapeutic applications for tissue regeneration. This review focuses mainly on the advances of natural and synthetic biomaterials in regulating stem cell fate decisions. We have also briefly discussed how biological and biophysical properties of biomaterials including wettability, chemical functionality, biodegradability and stiffness play their roles

Stimulus responsive graphene scaffolds for tissue engineering

Tissue engineering (TE) is an emerging area that aims to repair damaged tissues and organs by combining different scaffold materials with living cells. Recently, scientists started to engineer a new generation of nanocomposite scaffolds able to mimic biochemical and biophysical mechanisms to modulate the cellular responses promoting the restoration of tissue structure or function. Due to its unique electrical, topographical and chemical properties, graphene is a material that holds a great potential for TE, being already considered as one of the best candidates for accelerating and guiding stem cell differentiations. Although this is a promising field there are still some challenges to overcome, such as the efficient control of the differentiation of the stem cells, especially in graphene-based microenvironments. Hence, this chapter will review the existing research related to the ability of graphene and its derivatives (graphene oxide and reduced graphene oxide) to induce stem cell differentiation into diverse lineages when under the influence of electrical, mechanical, optical and topographic stimulations

The effects of graphene and mesenchymal stem cells in cutaneous wound healing and their putative action mechanism

Iwona Lasocka,1 Elżbieta Jastrzębska,2 Lidia Szulc-Dąbrowska,3 Michał Skibniewski,4 Iwona Pasternak,5 Marie Hubalek Kalbacova,6,7 Ewa M Skibniewska1 1Department of Animal Environment Biology, Faculty of Animal Science, Warsaw University of Life Sciences, Warsaw, Poland; 2Department of Medical Biotechnology, Faculty of Chemistry, Warsaw University of Technology, Warsaw, Poland; 3Department of Preclinical Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland; 4Department of Morphological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland; 5Faculty of Physics, Warsaw University of Technology, Warsaw, Poland; 6Institute of Pathological Physiology, 1st Faculty of Medicine, Charles University, Prague, Czech Republic; 7Biomedical Center, Faculty of Medicine in Pilsen, Charles University, Pilsen, Czech Republic Abstract: This study provides a review of the therapeutic potential of graphene dressing scaffolds and mesenchymal stem cells (MSCs) and their synergistic effects with respect to cutaneous wound healing. This study also considers their putative action mechanism based on the antibacterial, immunomodulating, angiogenic, matrix remodeling effects of materials belonging to the graphene family and MSCs during the wound healing process. In addition, this study discusses the cytocompatibility of graphene, its uses as a platform for skin substitutes, the properties it possesses with respect to providing protection against microbial invasion as well as strategies aimed at minimizing the chance of the occurrence of sepsis. MSCs are capable of secreting several factors that exert a therapeutic impact on reparative processes and tissue regeneration. In light of experiments conducted to date, graphene combined with MSCs appears to have the potential to enhance both the wound healing process and infection control at the injury site. Keywords: graphene, mesenchymal stem cells, wound, healin