Bio-Interfaces Engineering Using Laser-Based Methods for Controlled Regulation of Mesenchymal Stem Cell Response In Vitro

The controlled interfacial properties of materials and modulated behaviours of cells and biomolecules on their surface are the requirements in the development of a new generation of high-performance biomaterials for regenerative medicine applications. Roughness, chemistry and mechanics of biomaterials are all sensed by cells. Organization of the environment at the nano- and the microscale, as well as chemical signals, triggers specific responses with further impact on cell fate. Particularly, human mesenchymal stem cells (hMSCs) hold a great promise in both basic developmental biology studies and regenerative medicine, as progenitors of bone cells. Their fate can be affected by various key regulatory factors (e.g. soluble growth factors, intrinsic, extrinsic environmental factors) that can be delivered by a fabricated scaffold. For example, when cultured on engineered environments that reproduce the physical features of the bone, hMSCs express tissue-specific transcription factors and consequently undergo an osteogenic fate. Therefore, producing smart bio-interfaces with targeted functionalities represents the key point in effective use of hierarchically topographical and chemical bioplatforms. In this chapter, we review laser-based approaches (e.g. Matrix-Assisted Pulsed Laser Evaporation (MAPLE), Laser-Induced Forward Transfer (LIFT), laser texturing and laser direct writing) used for the design of bio-interfaces aimed at controlling stem cell behaviour in vitro

Smart Thermoresponsive Surfaces Based on pNIPAm Coatings and Laser Method for Biological Applications

Various applications within last decades such as bacterially resistant surfaces, soft robotics, drug delivery systems, sensors and tissue engineering are poised to feature the importance of the ability to control bio-interfacial interactions. An enhanced attention is dedicated to designing smart stimuli-responsive interfaces for DNA, drug delivery, protein and cell based applications. Within this context, the thermoresponsive materials, especially poly(N-isopropylacrylamide) (pNIPAm) have been intensively used in tissue engineering applications for a controlled detachment of proteins and cells with a minimum of invasive effect on protein and cell structural conformation. The properties of smart bio-interfaces can be controlled by its composition and polymer architecture. Therefore, appropriate methods for obtaining controlled coatings are necessary. Laser methods were successfully used in the last decades for obtaining controlled organic and inorganic coatings for various types of applications, from electronics to tissue engineering. Among these, Matrix-Assisted Pulsed Laser Evaporation (MAPLE) technique bring us a step forward to other laser methods by avoiding damage and photochemical decomposition of materials. In this chapter we describe materials and approaches used for design of smart bio-interfaces aimed at controlling protein and cells behavior in vitro, focusing MAPLE method for tuning coatings characteristics in relation with biological response

Graphene nanoplatelets-sericin surface-modified Gum alloy for improved biological response

In this study a “Gum Metal” titanium-based alloy, Ti-31.7Nb-6.21Zr-1.4Fe-0.16O, was synthesized by melting and characterized in order to evaluate its potential for biomedical applications. The results showed that the newly developed alloy presents a very high strength, high plasticity and a low Young\u27s modulus relative to titanium alloys currently used in medicine. For further bone implant applications, the newly synthesized alloy was surface modified with graphene nanoplatelets (GNP), sericin (SS) and graphene nanoplatelets/sericine (GNP–SS) composite films via Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique. The characterization of each specimen was monitored by scanning electron microscopy (SEM), atomic force microscopy (AFM), contact angle (CA) measurements, and Fourier Transform Infrared Spectroscopy (FTIR). The materials\u27 surface analyses suggested the successful coating of GNP, SS and GNP–SS onto the alloy surface. Additionally, the activities of pre-osteoblasts such as cell adhesion, cytoskeleton organization, cell proliferation and differentiation potentials exhibited on these substrates were investigated. Results showed that the GNP–SS-coated substrate significantly enhanced the growth and osteogenic differentiation of MC3T3-E1 cells when compared to bare and GNP-coated alloy. Collectively, the results show that GNP–SS surface-modified Gum alloy can modulate the bioactivity of the pre-osteoblasts holding promise for improved biological response in vivo

Fabrication of Hybrid Electrodes by Laser-Induced Forward Transfer for the Detection of Cu²⁺ Ions

Composites based on poly(3,4-ethylenedioxythiophene): poly(styrene sulfonate) (PEDOT:PSS)—graphene oxide (GO) are increasingly considered for sensing applications. In this work we aim at patterning and prototyping microscale geometries of PEDOT:PSS: GO composites for the modification of commercially available electrochemical sensors. Here, we demonstrate the laser-induced forward transfer of PEDOT:PSS: GO composites, a remarkably simple procedure that allows for the fast and clean transfer of materials with high resolution for a wide range of laser fluences (450–750 mJ/cm2). We show that it is possible to transfer PEDOT:PSS: GO composites at different ratios (i.e., 25:75 %wt and 50:50 %wt) onto flexible screen-printed electrodes. Furthermore, when testing the functionality of the PEDOT:PSS: GO modified electrodes via LIFT, we could see that both the PEDOT:PSS: GO ratio as well as the addition of an intermediate release layer in the LIFT process plays an important role in the electrochemical response. In particular, the ratio of the oxidation peak current to the reduction peak current is almost twice as high for the sensor with a 50:50 %et PEDOT:PSS: GO pixel. This direct transfer methodology provides a path forward for the prototyping and production of polymer: graphene oxide composite based devices

Material Erosion and Dust Formation during Tungsten Exposure to Hollow-Cathode and Microjet Discharges

International audienceTungsten erosion and dust occurrence are phenomena of great interest for fusion technology. Herein, we report results concerning the material damage and dust formation in the presence of high temperature and large area or concentrated discharges in helium and argon. In order to generate adequate plasmas, we used tungsten electrodes in two experimental discharge systems, namely a hollow discharge and a microjet discharge. In both exposure cases, we noticed surface modification, which was assigned to sputtering, melting, and vaporization processes, and a significant dust presence. We report the formation on electrode surfaces of tungsten fuzz, nano-cones, nanofibers, and cauliflower- and faced-like particles, depending on the discharge and gas type. Dust with various morphologies and sizes was collected and analyzed with respect to the morphology, size distribution, and chemical composition. We noticed, with respect to erosion and particle formation, common behaviors of W in both laboratory and fusion facilities experiments

Facile Fabrication of Hybrid Carbon Nanotube Sensors by Laser Direct Transfer

Ammonia is one of the most frequently produced chemicals in the world, and thus, reliable measurements of different NH3 concentrations are critical for a variety of industries, among which are the agricultural and healthcare sectors. The currently available technologies for the detection of NH3 provide accurate identification; however, they are limited by size, portability, and fabrication cost. Therefore, in this work, we report the laser-induced forward transfer (LIFT) of single-walled carbon nanotubes (SWCNTs) decorated with tin oxide nanoparticles (SnO2 NPs), which act as sensitive materials in chemiresistive NH3 sensors. We demonstrate that the LIFT-fabricated sensors can detect NH3 at room temperature and have a response time of 13 s (for 25 ppm NH3). In addition, the laser-fabricated sensors are fully reversible when exposed to multiple cycles of NH3 and have an excellent theoretical limit of detection of 24 ppt.ISSN:2079-499

New Poly(N-isopropylacrylamide-butylacrylate) Copolymer Biointerfaces and Their Characteristic Influence on Cell Behavior In Vitro

Designing and obtaining new synthetic smart biointerfaces with specific and controlled characteristics relevant for applications in biomedical and bioengineering domains represents one of the main challenges in these fields. In this work, Matrix-Assisted Pulsed Laser Evaporation (MAPLE) is used to obtain synthetic biointerfaces of poly(N-isopropyl acrylamide-butyl acrylate) p(NIPAM-BA) copolymer with different characteristics (i.e., roughness, porosity, wettability), and their effect on normal HEK 293 T and murine melanoma B16-F1 cells is studied. For this, the influence of various solvents (chloroform, dimethylsulfoxide, water) and fluence variation (250–450 mJ/cm2) on the morphological, roughness, wettability, and physico–chemical characteristics of the coatings are evaluated by atomic force microscopy, scanning electron microscopy, contact angle measurements, Fourier-transform-IR spectroscopy, and X-ray photoelectron spectroscopy. Coatings obtained by the spin coating method are used for reference. No significant alteration in the chemistry of the surfaces is observed for the coatings obtained by both methods. All p(NIPAM-BA) coatings show hydrophilic character, with the exception of those obtained with chloroform at 250 mJ/cm2. The surface morphology is shown to depend on both solvent type and laser fluence and it ranges from smooth surfaces to rough and porous ones. Physico–chemical and biological analysis reveal that the MAPLE deposition method with fluences of 350–450 mJ/cm2 when using DMSO solvent is more appropriate for bioengineering applications due to the surface characteristics (i.e., pore presence) and to the good compatibility with normal cells and cytotoxicity against melanoma cells

Human Mesenchymal Stem Cell Response to Lactoferrin-based Composite Coatings

The potential of mesenchymal stem cells (MSCs) for implantology and cell-based therapy represents one of the major ongoing research subjects within the last decades. In bone regeneration applications, the various environmental factors including bioactive compounds such as growth factors, chemicals and physical characteristics of biointerfaces are the key factors in controlling and regulating osteogenic differentiation from MSCs. In our study, we have investigated the influence of Lactoferrin (Lf) and Hydroxyapatite (HA) embedded within a biodegradable PEG-PCL copolymer on the osteogenic fate of MSCs, previous studies revealing an anti-inflammatory potential of the coating and osteogenic differentiation of murine pre-osteoblast cells. The copolymer matrix was obtained by the Matrix Assisted Pulsed Laser Evaporation technique (MAPLE) and the composite layers containing the bioactive compounds (Lf, HA, and Lf-HA) were characterised by Scanning Electron Microscopy and Atomic Force Microscopy. Energy-dispersive X-ray spectroscopy contact angle and surface energy of the analysed coatings were also measured. The characteristics of the composite surfaces were correlated with the viability, proliferation, and morphology of human MSCs (hMSCs) cultured on the developed coatings. All surfaces were found not to exhibit toxicity, as confirmed by the LIVE/DEAD assay. The Lf-HA composite exhibited an increase in osteogenic differentiation of hMSCs, results supported by alkaline phosphatase and mineralisation assays. This is the first report of the capacity of biodegradable composite layers containing Lf to induce osteogenic differentiation from hMSCs, a property revealing its potential for application in bone regeneration

Macrophage in vitro Response on Hybrid Coatings Obtained by Matrix Assisted Pulsed Laser Evaporation

The improvement in the research area of the implant by surface functionalization when correlated with the biological response is of major interest in the biomedical field. Based on the fact that the inflammatory response is directly involved in the ultimate response of the implant within the body, it is essential to study the macrophage-material interactions. Within this context, we have investigated the composite material-macrophage cell interactions and the inflammatory response to these composites with amorphous hydroxyapatite (HA), Lactoferrin (Lf), and polyethylene glycol-polycaprolactone (PEG-PCL) copolymer. All materials are obtained by Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique and characterized by Atomic Force Microscopy and Scanning Electron Microscopy. Macrophage-differentiated THP-1 cells proliferation and metabolic activity were assessed by qualitative and quantitative methods. The secretion of tumor necrosis factor alpha (TNF-α) and interleukin 10 (IL-10) cytokine, in the presence and absence of the inflammatory stimuli (bacterial endotoxin; lipopolysaccharide (LPS)), was measured using an ELISA assay. Our results revealed that the cellular response depended on the physical-chemical characteristics of the coatings. Copolymer-HA-Lf coatings led to low level of pro-inflammatory TNF-α, the increased level of anti-inflammatory IL-10, and the polarization of THP-1 cells towards an M2 pro-reparative phenotype in the presence of LPS. These findings could have important potential for the development of composite coatings in implant applications

Facile Fabrication of Hybrid Carbon Nanotube Sensors by Laser Direct Transfer

Ammonia is one of the most frequently produced chemicals in the world, and thus, reliable measurements of different NH3 concentrations are critical for a variety of industries, among which are the agricultural and healthcare sectors. The currently available technologies for the detection of NH3 provide accurate identification; however, they are limited by size, portability, and fabrication cost. Therefore, in this work, we report the laser-induced forward transfer (LIFT) of single-walled carbon nanotubes (SWCNTs) decorated with tin oxide nanoparticles (SnO2 NPs), which act as sensitive materials in chemiresistive NH3 sensors. We demonstrate that the LIFT-fabricated sensors can detect NH3 at room temperature and have a response time of 13 s (for 25 ppm NH3). In addition, the laser-fabricated sensors are fully reversible when exposed to multiple cycles of NH3 and have an excellent theoretical limit of detection of 24 ppt