Revealing bactericidal events on graphene oxide nano films deposited on metal implant surfaces.

At the time when pathogens are developing robust resistance to antibiotics, the demand for implant surfaces with microbe-killing capabilities has significantly risen. To achieve this goal, profound understanding of the underlying mechanisms is crucial. Our study demonstrates that graphene oxide (GO) nano films deposited on stainless steel (SS316L) exhibit superior antibacterial features. The physicochemical properties of GO itself play a pivotal role in influencing biological events and their diversity may account for the contradictory results reported elsewhere. However, essential properties of GO coatings, such as oxygen content and the resulting electrical conductivity, have been overlooked so far. We hypothesize that the surface potential and electrical resistance of the oxygen content in the GO-nano films may induce bacteria-killing events on conductive metallic substrates. In our study, the GO applied contains 52 wt% of oxygen, and thus exhibits insulating properties. When deposited as a nano film on an electrically conducting steel substrate, GO flakes generate a Schottky barrier at the interface. This barrier, consequently, impedes the transfer of electrons to the underlying conductive substrate. As a result, this creates reactive oxygen species (ROS), leading to bacterial death. We confirmed the presence of GO coatings and their hydrolytic stability by using X-ray photoelectron spectroscopy (XPS), µRaman spectroscopy, scanning electron microscopy (SEM), and Kelvin probe force microscopy (KPFM) measurements. The biological evaluation was performed on the MG63 osteoblast-like cell line and two selected bacteria species: S. aureus and P. aeruginosa, demonstrating both the cytocompatibility and antibacterial behavior of GO-coated SS316L substrates. We propose a two-step bactericidal mechanism: electron transfer from the bacteria membrane to the substrate, followed by ROS generation. This mechanism finds support in changes observed in contact angle, surface potential, and work function, identified as decisive factors. By addressing overlooked factors and effectively bridging the gap between understanding and practicality, we present a transformative approach for implant surfaces, combating microbial resistance, and offering new application possibilitie.

Deep eutectic solvent-assisted fabrication of bioinspired 3D carbon-calcium phosphate scaffolds for bone tissue engineering.

Tissue engineering is a burgeoning field focused on repairing damaged tissues through the combination of bodily cells with highly porous scaffold biomaterials, which serve as templates for tissue regeneration, thus facilitating the growth of new tissue. Carbon materials, constituting an emerging class of superior materials, are currently experiencing remarkable scientific and technological advancements. Consequently, the development of novel 3D carbon-based composite materials has become significant for biomedicine. There is an urgent need for the development of hybrids that will combine the unique bioactivity of ceramics with the performance of carbonaceous materials. Considering these requirements, herein, we propose a straightforward method of producing a 3D carbon-based scaffold that resembles the structural features of spongin, even on the nanometric level of their hierarchical organization. The modification of spongin with calcium phosphate was achieved in a deep eutectic solvent (choline chloride : urea, 1 : 2). The holistic characterization of the scaffolds confirms their remarkable structural features (i.e., porosity, connectivity), along with the biocompatibility of α-tricalcium phosphate (α-TCP), rendering them a promising candidate for stem cell-based tissue-engineering. Culturing human bone marrow mesenchymal stem cells (hMSC) on the surface of the biomimetic scaffold further verifies its growth-facilitating properties, promoting the differentiation of these cells in the osteogenesis direction. ALP activity was significantly higher in osteogenic medium compared to proliferation, indicating the differentiation of hMSC towards osteoblasts. However, no significant difference between C and C-αTCP in the same medium type was observed.

Flake Graphene as an Innovative Additive to Grease with Improved Tribological Properties.

The paper presents the results of research on the use of flake graphene as an additive to plastic grease in order to improve its tribological properties. The influence of concentration (0.25-5.00 wt.%) and the form of graphene (graphene oxide, reduced graphene oxide) on selected properties of the base grease were investigated. It has been found that the addition of graphene flakes improves the anti-wear properties of the lubricant. The greatest improvement in the properties of the lubricant was achieved by using graphene at a concentration of 4.00 wt.%; the reduction in the average diameter of the wear scar was almost 70% for GO and RGO, compared to the base lubricant without the addition of graphene.

How Streptococcus mutans Affects the Surface Topography and Electrochemical Behavior of Nanostructured Bulk Ti.

The metabolization of carbohydrates by Streptococcus mutans leads to the formation of lactic acid in the oral cavity, which can consequently accelerate the degradation of dental implants fabricated from commercially available microcrystalline Ti. Microstructure influences surface topography and hence interaction between bacteria cells and Ti surfaces. This work offers the first description of the effect of S. mutans on the surface topography and properties of nanostructured bulk Ti, which is a promising candidate for modern narrow dental implants owing to its superior mechanical strength. It was found that S. mutans incubation resulted in the slight, unexpected decrease of surface nanoroughness, which was previously developed owing to privileged oxidation in areas of closely spaced boundaries. However, despite the changes in nanoscale surface topography, bacteria incubation did not reduce the high level of protection afforded by the oxide layer formed on the nanostructured Ti surface. The results highlight the need-hitherto ignored-to consider Ti microstructure when analyzing its behavior in the presence of carbohydrate-metabolizing bacteria.

Flake Graphene as an Efficient Agent Governing Cellular Fate and Antimicrobial Properties of Fibrous Tissue Engineering Scaffolds-A Review.

Although there are several methods for fabricating nanofibrous scaffolds for biomedical applications, electrospinning is probably the most versatile and feasible process. Electrospinning enables the preparation of reproducible, homogeneous fibers from many types of polymers. In addition, implementation of this technique gives the possibility to fabricated polymer-based composite mats embroidered with manifold materials, such as graphene. Flake graphene and its derivatives represent an extremely promising material for imparting new, biomedically relevant properties, functions, and applications. Graphene oxide (GO) and reduced graphene oxide (rGO), among many extraordinary properties, confer antimicrobial properties of the resulting material. Moreover, graphene oxide and reduced graphene oxide promote the desired cellular response. Tissue engineering and regenerative medicine enable advanced treatments to regenerate damaged tissues and organs. This review provides a reliable summary of the recent scientific literature on the fabrication of nanofibers and their further modification with GO/rGO flakes for biomedical applications.

Flake Graphene-Based Nanomaterial Approach for Triggering a Ferroptosis as an Attractive Theranostic Outlook for Tackling Non-Small Lung Cancer: A Mini Review.

Lung cancer is a highly aggressive neoplasm that is now a leading cause of cancer death worldwide. One of the major approaches for killing cancer cells is related with activation of apoptotic cell death with anti-cancer drugs. However, the efficiency of apoptosis induction in tumors is limited. Consequently, the development of other forms of non-apoptotic cell death is up to date challenge for scientists worldwide. This situation motivated us to define the aim of this mini-review: gathering knowledge regarding ferroptosis-newly defined programmed cell death process characterized by the excessive accumulation of iron-and combining it with yet another interesting nanomaterial-based graphene approach. In this manuscript, we presented brief information about non-small lung cancer and ferroptosis, followed by a section depicting the key-features of graphene-based nanomaterials influencing their biologically relevant properties.

Luminescence Properties of Nano Zinc Oxide Doped with Al(III) Ions Obtained in Microwave-Assisted Hydrothermal Synthesis.

The hydrothermal method of obtaining nano zinc oxide doped with different contents of aluminum ions (III) was presented and discussed in this paper. Aqueous solution of Zn(NO3)2*6H2O and Al(NO3)3*9H2O salts mixture were used as the synthesis precursor. In order to reduce the process time all reactions were performed in a microwave reactor. The influence of process parameters and the content of impurity ions on the properties of synthesized nano zinc oxide were analyzed. In addition to zinc oxide doped with Al(III) ions, an additional spinel phase (ZnAl2O4) was obtained. The luminescent properties of nano zinc oxide as a function of the dopant ions were also discussed. Based on the luminescence measurements results, it was found that the luminescence intensity decreases with the increasing dopant content. The obtained materials are aimed to be implemented as luminescent materials in optoelectronic and sensors.

Naturally prefabricated 3D chitinous skeletal scaffold of marine demosponge origin, biomineralized ex vivo as a functional biomaterial.

Solutions developed by nature for structural and functional optimization of three-dimensional (3D) skeletal structures provide unique windows not only into the evolutionary pathways of organisms, but also into bioinspired materials science and biomimetics. Great examples are naturally formed 3D chitinous scaffolds of marine sponge remain a focus of modern biomedicine and tissue engineering. Due to its properties like renewability, bioactivity, and biodegradability such constructs became very interesting players as components of organic-inorganic biocomposites. Herein, we developed chitin-based biocomposites by biomimetic ex vivo deposition of calcium carbonate particles using hemolymph from the cultivated mollusk Cornu aspersum and chitinous matrix from the marine demosponge Aplysina fistularis. The biological potential of the developed biofunctionalized scaffolds for bone tissue engineering was evaluated by investigating the spreading and viability of a human fetal osteoblast cell line has been determined for the first time. Performed analyses like dynamic mechanical analysis and atomic force microscopy shown that biofunctionalized scaffold possess about 4 times higher mechanical resistance. Moreover, several topographical changes have been observed, as e.g., surface roughness (Rq) increased from 31.75 ± 2.7 nm to 120.7 ± 0.3 nm. The results are indicating its potential for use in the modification of cell delivery systems in future biomedical applications.

Morphology and Chemical Purity of Water Suspension of Graphene Oxide FLAKES Aged for 14 Months in Ambient Conditions. A Preliminary Study.

Graphene and its derivatives have attracted scientists' interest due to their exceptional properties, making them alluring candidates for multiple applications. However, still little is known about the properties of as-obtained graphene derivatives during long-term storage. The aim of this study was to check whether or not 14 months of storage time impacts graphene oxide flakes' suspension purity. Complementary micro and nanoscale characterization techniques (SEM, AFM, EDS, FTIR, Raman spectroscopy, and elemental combustion analysis) were implemented for a detailed description of the topography and chemical properties of graphene oxide flakes. The final step was pH evaluation of as-obtained and aged samples. Our findings show that purified flakes sustained their purity over 14 months of storage.

Water vapor induced self-assembly of islands/honeycomb structure by secondary phase separation in polystyrene solution with bimodal molecular weight distribution.

The formation of complex structures in thin films is of interest in many fields. Segregation of polymer chains of different molecular weights is a well-known process. However, here, polystyrene with bimodal molecular weight distribution, but no additional chemical modification was used. It was proven that at certain conditions, the phase separation occurred between two fractions of bimodal polystyrene/methyl ethyl ketone solution. The films were prepared by spin-coating, and the segregation between polystyrene phases was investigated by force spectroscopy. Next, water vapour induced secondary phase separation was investigated. The introduction of moist airflow induced the self-assembly of the lower molecular weight into islands and the heavier fraction into a honeycomb. As a result, an easy, fast, and effective method of obtaining island/honeycomb morphologies was demonstrated. The possible mechanisms of the formation of such structures were discussed.

Investigation into morphological and electromechanical surface properties of reduced-graphene-oxide-loaded composite fibers for bone tissue engineering applications: A comprehensive nanoscale study using atomic force microscopy approach.

We decided to implement an extensive atomic force microscopy study in order to get deeper understanding of surface-related nanoscale properties of 3D printed pristine polycaprolactone and its reduced-graphene-oxide-loaded composites. The study included surface visualization and roughness quantification, elastic modulus and adhesion force assessment with force spectroscopy, along with kelvin probe force microscopy evaluation of local changes of surface potential. Atomic force microscopy examination was followed by scanning electron microscopy visualization and wettability assessment. Moreover, systematic examination of reduced graphene oxide flakes fabricated exclusively for this study was performed, including: scanning electron microscopy, Raman spectroscopy, X-ray photoelectron spectroscopy and combustion elemental analysis. The addition of reduced graphene oxide resulted in thickening of the composite fibers and surface roughness enhancement. In addition, elastic modulus of composite fibers was higher and at the same time adhesion forces between scanning probe and tested surface was lower than for pristine polymeric ones. Lastly, we recorded local (nanoscale) alterations of surface potential of fibers with addition of graphene-derivative. The results clearly suggest graphene derivative's dose-dependent alteration of elastic modulus and adhesion force recorded with atomic force microscope. Moreover, changes of the material's surface properties were followed by changes of its electrical properties.

Microwave-Assisted Hydrothermal Synthesis of Zinc-Aluminum Spinel ZnAl2O4.

The drawback of the hydrothermal technique is driven by the fact that it is a time-consuming operation, which greatly impedes its commercial application. To overcome this issue, conventional hydrothermal synthesis can be improved by the implementation of microwaves, which should result in enhanced process kinetics and, at the same time, pure-phase and homogeneous products. In this study, nanometric zinc aluminate (ZnAl2O4) with a spinel structure was obtained by a hydrothermal method using microwave reactor. The average ZnAl2O4 crystallite grain size was calculated from the broadening of XRD lines. In addition, BET analysis was performed to further characterize the as-synthesized particles. The synthesized materials were also subjected to microscopic SEM and TEM observations. Based on the obtained results, we concluded that the grain sizes were in the range of 6-8 nm. The surface areas measured for the samples from the microwave reactor were 215 and 278 m2 g-1.

From Matrix Vesicles to Miniature Rocks: Evolution of Calcium Deposits in Calf Costochondral Junctions.

OBJECTIVE: Initial stages of cartilage matrix calcification depend on the activity of matrix vesicles. The purpose of the study was to describe how calcified matrix vesicles join into larger structures, to present their up-to-date undescribed 3-dimensional image, and to observe how calcified matrix relates to chondrocyte lacunae. DESIGN: Calcified cartilage was obtained from the zone of provisional calcification of calf costochondral junctions, then enzymatically isolated and studied by microtomography, scanning electron microscopy, atomic force microscopy and X-ray diffraction, and Fourier transform infrared spectroscopy. RESULTS: Hyaluronidase digestion released packets of granules surrounded by the cartilage matrix. Further digestion, with collagenase and trypsin, removed matrix and exposed granules with dimensions within 50 to 150 nm range, which we consider as equivalent of calcified matrix vesicles. Granules joined into larger groups with dimensions of 0.5 to 2 µm, which we call globular units. Certain matrix vesicles appeared well connected but contained globular units that had spaces filled with electron lucent material, presumably matrix or chondrocyte remnants. Globular units were organized into massive structures taking the shape of oval plates. Comparison of these plates with lacunae containing isogenous groups of chondrocytes from proliferative zone of costochondral junction suggests that the cells from a single lacuna were responsible for the formation of one plate. The plates were connected with each other and extended over provisional calcification zone. CONCLUSIONS: The outcome showed how particular calcified matrix vesicles associate into globular units, which organize into massive structures assuming the shape of oval plates and eventually cover large areas of cartilage matrix.

The effect of diameter of fibre on formation of hydrogen bonds and mechanical properties of 3D-printed PCL.

Fused Deposition Modelling (FDM) technique has been widely utilized in fabrication of 3D porous scaffolds for tissue engineering (TE) applications. Surprisingly, although there are many publications devoted to the architectural features of the 3D scaffolds fabricated by the FDM, none of them give us evident information about the impact of the diameter of the fibres on material properties. Therefore, the aim of this study was to investigate, for the first time, the effect of the diameter of 3D-printed PCL fibres on variations in their microstructure and resulting mechanical behaviour. The fibres made of poly(ε-caprolactone) (PCL) were extruded through commonly used types of nozzles (inner diameter ranging from 0.18 mm to 1.07 mm) by means of FDM technique. Static tensile test and atomic force microscopy working in force spectroscopy mode revealed strong decrease in the Young's modulus and yield strength with increasing fibre diameter in the investigated range. To explain this phenomenon, we conducted differential scanning calorimetry, wide-angle X-ray-scattering, Fourier-transform infrared spectroscopy, infrared and polarized light microscopy imaging. The obtained results clearly showed that the most prominent effect on the obtained microstructures and mechanical properties had different cooling and shear rates during fabrication process causing changes in supramolecular interactions of PCL. The observed fibre size-dependent formation of hydrogen bonds affected the crystalline structure and its stability. Summarising, this study clearly demonstrates that the diameter of 3D-printed fibres has a strong effect on obtained microstructure and mechanical properties, therefore should be taken into consideration during design of the 3D TE scaffolds.

Synthesis and Characterization of Graphene Oxide and Reduced Graphene Oxide Composites with Inorganic Nanoparticles for Biomedical Applications.

Graphene oxide (GO) and reduced graphene oxide (RGO), due to their large active surface areas, can serve as a platform for biological molecule adhesion (both organic and inorganic). In this work we described methods of preparing composites consisting of GO and RGO and inorganic nanoparticles of specified biological properties: nanoAg, nanoAu, nanoTiO2 and nanoAg2O. The idea of this work was to introduce effective methods of production of these composites that could be used for future biomedical applications such as antibiotics, tissue regeneration, anticancer therapy, or bioimaging. In order to characterize the pristine graphene materials and resulting composites, we used spectroscopic techniques: XPS and Raman, microscopic techniques: SEM with and AFM, followed by X-Ray diffraction. We obtained volumetric composites of flake graphene and Ag, Au, Ag2O, and TiO2 nanoparticles; moreover, Ag nanoparticles were obtained using three different approaches.

Internal nanocrystalline structure and stiffness alterations of electrospun polycaprolactone-based mats after six months of in vitro degradation. An atomic force microscopy assay.

Biodegradable electrospun nanofibrous scaffolds for bone tissue engineering applications have been extensively studied as they can provide attractive open-worked architecture resembling natural extracellular matrix, with tunable physical and mechanical properties enhancing positive cellular response. For this purpose, electrospun mats were tested in terms of morphology, mechanical and physical properties, degradation kinetics and related phenomena occurring in micro- and nanoscale. However, detailed description of internal nanostructures of electrospun mats and their changes related to in vitro degradation is still missing. In this manuscript, we report qualitative and quantitative evaluation of internal lamellar nanostructure of electrospun fibrous scaffolds made of pristine polycaprolactone and composite with polymeric matrix and nanoceramic (hydroxyapatite) filler during in vitro degradation. Morphological and mechanical studies performed with an atomic force microscope were followed by scanning electron microscope imaging and X-Ray diffraction. The results suggest degradation-dependent alteration of both organization and thickness of nano-scaled lamellas recorded with atomic force microscope. Moreover, changes of the material's internal structure were followed by enhanced stiffness and higher crystallinity of electrospun fibers.

In vivo and in vitro study of a novel nanohydroxyapatite sonocoated scaffolds for enhanced bone regeneration.

There still remains a need for new methods of healing large bone defects, i.e., gaps in bone tissue that are too big to naturally heal. Bone regrowth scaffolds can fill the bone gap and enhance the bone regeneration by providing cells with a support to for new tissue formation. Coating of the scaffolds surface with nanocrystalline hydroxyapatite may enhance the osteoinductivity or osteoconductivity of such scaffolds. Here we present the sonocoating method to coat scaffolds with bioactive hydroxyapatite nanoparticles. We show a method, where the material to be coated is immersed in a colloidal suspension of nanoparticles with mean sizes of 10 nm and 43 nm in water, and high-power ultrasound waves are applied to the suspension for 15 min at 30 °C. High power ultrasounds lead to growth of cavitation bubbles in liquid, which implode at a critical size. The implosion energy propels the nanoparticles towards the material surface, causing their attachment to the scaffold. Using this technique, we produced a uniform layer of nanohydroxyapatite particles of thickness in the range 200 to 300 nm on two types of scaffolds: a porous ß-TCP ceramic scaffold and a 3D-printed scaffold made of PCL fibers. In vivo tests in rabbits confirmed that the novel coating strongly stimulated new bone tissue formation, with new bone tissue occupying 33% for the nHAP-coated PCL scaffold and 68% for the nHAP-coated ß-TCP after a 3-month test. The sonocoating method leads to formation of a bioactive layer on the scaffolds at temperature close to room temperature, very short time and in water. It is a green technological process, promising for bone tissue regeneration applications.

Binary bioactive glass composite scaffolds for bone tissue engineering-Structure and mechanical properties in micro and nano scale. A preliminary study.

Composite scaffolds of bioactive glass (SiO2-CaO) and bioresorbable polyesters: poly-l-lactic acid (PLLA) and polycaprolactone (PCL) were produced by polymer coating of porous foams. Their structure and mechanical properties were investigated in micro and nanoscale, by the means of scanning electron microscopy, PeakForce Quantitative Nanomechanical Property Mapping (PF-QNM) atomic force microscopy, micro-computed tomography and contact angle measurements. This is one of the first studies in which the nanomechanical properties (elastic modulus, adhesion) were measured and mapped simultaneously with topography imaging (PF-QNM AFM) for bioactive glass and bioactive glass - polymer coated scaffolds. Our findings show that polymer coated scaffolds had higher average roughness and lower stiffness in comparison to pure bioactive glass scaffolds. Such coating-dependent scaffold properties may promote different cells-scaffold interaction.

Surface Modification of 3D Printed Polycaprolactone Constructs via a Solvent Treatment: Impact on Physical and Osteogenic Properties.

One promising strategy to reconstruct bone defects relies on 3D printed porous structures. In spite of several studies having been carried out to fabricate controlled, interconnected porous constructs, the control over surface features at, or below, the microscopic scale remains elusive for 3D polymeric scaffolds. In this study, we developed and refined a methodology which can be applied to homogeneously and reproducibly modify the surface of polymeric 3D printed scaffolds. We have demonstrated that the combination of a polymer solvent and the utilization of ultrasound was essential for achieving appropriate surface modification without damaging the structural integrity of the construct. The modification created on the scaffold profoundly affected the macroscopic and microscopic properties of the scaffold with an increased roughness, greater surface area, and reduced hydrophobicity. Furthermore, to assess the performance of such materials in bone tissue engineering, human mesenchymal stem cells (hMSC) were cultured in vitro on the scaffolds for up to 7 days. Our results demonstrate a stronger commitment toward early osteogenic differentiation of hMSC. Finally, we demonstrated that the increased in the specific surface area of the scaffold did not necessarily correlate with improved adsorption of protein and that other factors, such as surface chemistry and hydrophilicity, may also play a major role.

Fabrication, multi-scale characterization and in-vitro evaluation of porous hybrid bioactive glass polymer-coated scaffolds for bone tissue engineering.

Bioactive glass-based scaffolds are commonly used in bone tissue engineering due to their biocompatibility, mechanical strength and adequate porous structure. However, their hydrophobicity and brittleness limits their practical application. In this study, to improve nanomechanical properties of such scaffolds, pure bioactive hybrid glass and two bioactive hybrid glass-polymer coated composites were fabricated. A complementary micro and nanoscale characterization techniques (SEM, AFM, µCT, FTIR, compressive test, goniometer) were implemented for detailed description of architecture and physicochemical properties of hybrid bioactive glass-based scaffolds with emphasis on nano-mechanics. The final step was in-vitro evaluation of three dimensional macroporous structures. Our findings show that after polymer addition, architecture, topography and surface properties of the scaffolds were changed and promoted favoured behaviour of the cells.