In vivo biocompatibility of diamond-like carbon films containing TiO2 nanoparticles for biomedical applications.

Hybrid diamond-like carbon (DLC) with incorporated titanium dioxide (TiO2) nanoparticle coatings have low friction coefficient, high wear resistance, high hardness, biocompatibility, and high chemical stability. They could be employed to modify biomedical alloys surfaces for numerous applications in biomedical engineering. Here we investigate for the first time the in vivo inflammatory process of DLC coatings with incorporated TiO2 nanoparticles. TiO2-DLC films were grown on AISI 316 stainless-steel substrates using plasma-enhanced chemical vapor deposition. The coated substrates were implanted in CF1 mice peritoneum. The in vivo cytotoxicity and biocompatibility of the samples were analyzed from macrophage lavage. Analysis in the first weeks after implantation could be helpful to evaluate the acute cytotoxicity generated after a possible inflammatory process. The in vivo results showed no inflammatory process. A significant increase in nitric oxide production on the uncoated substrates was confirmed through cytometry, and the coated substrates demonstrated biocompatibility. The presence of TiO2 nanoparticles enhanced the wound healing activity, due to their astringent and antimicrobial properties. DLC and TiO2-DLC coatings were considered biocompatible, and the presence of TiO2 nanoparticles reduced the inflammatory reactions, increasing DLC biocompatibility.

A comparison between electrospinning and rotary-jet spinning to produce PCL fibers with low bacteria colonization.

One of the important components in tissue engineering is material structure, providing a model for fixing and the development of cells and tissues, which allows for the transport of nutrients and regulatory molecules to and from cells. The community claims the need for new materials with better properties for use in the clinic. Poly (ε-caprolactone) (PCL) is a biodegradable polymer, semi crystalline, with superior mechanical properties and has attracted an increasing interest due to its usefulness in various biomedical applications. Herein, two different methods (electrospinning versus rotary jet spinning) with different concentrations of PCL produced ultra thin-fibers each with particular characteristics, verified and analyzed by morphology, wettability, thermal and cytotoxicity features and for bacteria colonization. Different PCL scaffold morphologies were found to be dependent on the fabrication method used. All PCL scaffolds showed greater mammalian cell interactions. Most impressively, rotary-jet spun fibers showed that a special rough surface decreased bacteria colonization, emphasizing that no nanoparticle or antibiotic was used; maybe this effect is related with physical (scaffold) and/or biological mechanisms. Thus, this study showed that rotary jet spun fibers possess a special topography compared to electrospun fibers to reduce bacteria colonization and present no cytotoxicity when in contact with mammalian cells.

Biomineralized diamond-like carbon films with incorporated titanium dioxide nanoparticles improved bioactivity properties and reduced biofilm formation.

Recently, the development of coatings to protect biomedical alloys from oxidation, passivation and to reduce the ability for a bacterial biofilm to form after implantation has emerged. Diamond-like carbon films are commonly used for implanted medical due to their physical and chemical characteristics, showing good interactions with the biological environment. However, these properties can be significantly improved when titanium dioxide nanoparticles are included, especially to enhance the bactericidal properties of the films. So far, the deposition of hydroxyapatite on the film surface has been studied in order to improve biocompatibility and bioactive behavior. Herein, we developed a new route to obtain a homogeneous and crystalline apatite coating on diamond-like carbon films grown on 304 biomedical stainless steel and evaluated its antibacterial effect. For this purpose, films containing two different concentrations of titanium dioxide (0.1 and 0.3g/L) were obtained by chemical vapor deposition. To obtain the apatite layer, the samples were soaked in simulated body fluid solution for up to 21days. The antibacterial activity of the films was evaluated by bacterial eradication tests using Staphylococcus aureus biofilm. Scanning electron microscopy, X-ray diffraction, Raman scattering spectroscopy, and goniometry showed that homogeneous, crystalline, and hydrophilic apatite films were formed independently of the titanium dioxide concentration. Interestingly, the diamond-like films containing titanium dioxide and hydroxyapatite reduced the biofilm formation compared to controls. A synergism between hydroxyapatite and titanium dioxide that provided an antimicrobial effect against opportunistic pathogens was clearly observed.

Graphene oxide nanoribbons as nanomaterial for bone regeneration: Effects on cytotoxicity, gene expression and bactericidal effect.

Graphene oxide nanoribbons (O-GNR) surges as an interesting nanomaterial for biomedical applications due to feasibility to incorporate functional groups and possible bactericidal properties. Herein, high concentrations of O-GNR were biologically evaluated using human osteoblast cells and gram positive and negative bacteria. Briefly, our goal were to evaluate: (1) synthetic pathway, (2) characterization and (3) effects of O-GNR composition and structural factors as a new approach for biomedical applications. For this, O-GNR were produced combining chemical vapor deposition and oxygen plasma treatment of multiwalled carbon nanotubes. Then, we analyzed the bioactivity, cell viability, osteogenic differentiation, matrix mineralization, mRNA levels of the five genes related direct to bone repair and bactericidal effect of high concentrations of O-GNR (10µgmL-1, 100µgmL-1, 200µgmL-1 and 300µgmL-1). Impressively, O-GNR showed no cytotoxic effects up to a concentration of 100µgmL-1 and no gene expression alteration when used in its dose. We also observed that S. aureus and E. coli bacteria are susceptible to damage when incubated with 100µgmL-1 of O-GNR, showing approximately 50% of bacterial death. We consider that O-GNR displays attractive properties when used at a suitable dose, displaying bactericidal effect and apparently lacking to cause damages in the bone repair process.

Nanostructured poly (lactic acid) electrospun fiber with high loadings of TiO2 nanoparticles: Insights into bactericidal activity and cell viability.

Researchers have been looking for modifying surfaces of polymeric biomaterials approved by FDA to obtain nanofeatures and bactericidal properties. If modified, it would be very interesting because the antibiotic administration could be reduced and, therefore, the bacterial resistance. Here, we report the electrospinning of poly (lactic acid) (PLA) with high loadings of titanium dioxide nanoparticles (TiO2, 1-5wt%) and their bactericidal properties. TiO2 nanoparticles have been recognized for a long time for their antibacterial, low cost and self-cleaning properties. However, their ability to reduce bacteria functions when used in polymers has not been well studied to date. In this context, we aimed here to generate nanostructured PLA electrospun fiber-TiO2 nanoparticle composites for further evaluation of their bactericidal activity and cell viability. TEM and SEM micrographs revealed the successful electrospinning of PLA/TiO2 and the generation of polymer-TiO2 nanostructures. When increasing the TiO2 concentration, we observed a proportional increase in the nanoparticle density along the fiber and surface. The nanostructured PLA/TiO2 nanofibers showed no mammalian cell toxicity and, most importantly, possessed bactericidal activity with higher TiO2 loads. Such results suggest that the present PLA electrospun fiber-TiO2 nanoparticle composites should be further studied for a wide range of biomedical applications.

Assisted deposition of nano-hydroxyapatite onto exfoliated carbon nanotube oxide scaffolds.

Electrodeposited nano-hydroxyapatite (nHAp) is more similar to biological apatite in terms of microstructure and dimension than apatites prepared by other processes. Reinforcement with carbon nanotubes (CNTs) enhances its mechanical properties and increases adhesion of osteoblasts. Here, we carefully studied nHAp deposited onto vertically aligned multi-walled CNT (VAMWCNT) scaffolds by electrodeposition and soaking in a simulated body fluid (SBF). VAMWCNTs are porous biocompatible scaffolds with nanometric porosity and exceptional mechanical and chemical properties. The VAMWCNT films were prepared on a Ti substrate by a microwave plasma chemical vapour deposition method, and then oxidized and exfoliated by oxygen plasma etching (OPE) to produce graphene oxide (GO) at the VAMWCNT tips. The attachment of oxygen functional groups was found to be crucial for nHAp nucleation during electrodeposition. A thin layer of plate-like and needle-like nHAp with high crystallinity was formed without any need for thermal treatment. This composite (henceforth referred to as nHAp-VAMWCNT-GO) served as the scaffold for in vitro biomineralization when soaked in the SBF, resulting in the formation of both carbonate-rich and carbonate-poor globular-like nHAp. Different steps in the deposition of biological apatite onto VAMWCNT-GO and during the short-term biomineralization process were analysed. Due to their unique structure and properties, such nano-bio-composites may become useful in accelerating in vivo bone regeneration processes.

Analysis of cellular adhesion on superhydrophobic and superhydrophilic vertically aligned carbon nanotube scaffolds.

We analyzed GFP cells after 24h cultivated on superhydrophilic vertically aligned carbon nanotube scaffolds. We produced two different densities of VACNT scaffolds on Ti using Ni or Fe catalysts. A simple and fast oxygen plasma treatment promoted the superhydrophilicity of them. We used five different substrates, such as: as-grown VACNT produced using Ni as catalyst (Ni), as-grown VACNT produced using Fe as catalyst (Fe), VACNT-O produced using Ni as catalyst (NiO), VACNT-O produced using Fe as catalyst (FeO) and Ti (control). The 4',6-diamidino-2-phenylindole reagent nuclei stained the adherent cells cultivated on five different analyzed scaffolds. We used fluorescence microscopy for image collect, ImageJ® to count adhered cell and GraphPad Prism 5® for statistical analysis. We demonstrated in crescent order: Fe, Ni, NiO, FeO and Ti scaffolds that had an improved cellular adhesion. Oxygen treatment associated to high VACNT density (group FeO) presented significantly superior cell adhesion up to 24h. However, they do not show significant differences compared with Ti substrates (control). We demonstrated that all the analyzed substrates were nontoxic. Also, we proposed that the density and hydrophilicity influenced the cell adhesion behavior.

Graphene and carbon nanotube composite enabling a new prospective treatment for trichomoniasis disease.

We report the synthesis and application of novel graphene oxide and carbon nanotube oxide (GCN-O) composite. First, pristine multi-walled carbon nanotube was prepared by chemical vapour deposition furnace and then exfoliated and oxidised simultaneously by oxygen plasma etching. The superficial and volumetric compositions of GCN-O were measured by XPS spectroscopy and EDX spectroscopy, respectively. Both XPS and EDX analyses evidence that the GCN-O is composed of up to 20% of oxygen atoms. As a result, GCN-O forms a stable colloidal aqueous solution and shows to have strong interaction with the cell membrane of Tritrichomonas foetus protozoa, making easy its application as a drug carrier. Trichomoniasis infection of cattle is a devastating disease for cattle producers, causing some damages to females and fetus, and the abortion is the most serious result of this disease. There is no effective treatment for trichomoniasis infection yet. Therefore, new treatment, especially one with no collateral effects in animals, is required. With this goal in mind, our results suggest that water dispersible composite is a novel nanomaterial, promising for Trichomoniasis infection treatment and as therapeutic delivery agent as well.

Improvement of diamond-like carbon electrochemical corrosion resistance by addition of nanocrystalline diamond.

Nanocrystalline diamond (NCD) particles were incorporated into diamond-like carbon (DLC) films in order to investigate NCD-DLC electrochemical corrosion resistance. The films were grown over 304 stainless steel using plasma-enhanced chemical vapor deposition. NCD particles were incorporated into DLC during the deposition process. The investigation of NCD-DLC electrochemical corrosion behavior was performed using potentiodynamic polarization against NaCl. NCD-DLC films presented more negative corrosion potential and lower anodic and cathodic current densities. The electrochemical analysis indicated that NCD-DLC films present superior impedance and polarization resistance compared to the pure DLC, which indicate that they are promising corrosion protective coatings in aggressive solutions.

Antibacterial activity of DLC films containing TiO2 nanoparticles.

Diamond-like carbon (DLC) films have been the focus of extensive research in recent years due to their potential applications as surface coatings on biomedical devices. Titanium dioxide (TiO2) in the anatase crystalline form is a strong bactericidal agent when exposed to near-UV light. In this work we investigate the bactericidal activity of DLC films containing TiO2 nanoparticles. The films were grown on 316L stainless-steel substrates from a dispersion of TiO2 in hexane using plasma-enhanced chemical vapor deposition. The composition, bonding structure, surface energy, stress, and surface roughness of these films were also evaluated. The antibacterial tests were performed against Escherichia coli (E. coli) and the results were compared to the bacterial adhesion force to the studied surfaces. The presence of TiO2 in DLC bulk was confirmed by Raman spectroscopy. As TiO2 content increased, I(D)/I(G) ratio, hydrogen content, and roughness also increased; the films became more hydrophilic, with higher surface free energy and the interfacial energy of bacteria adhesion decreased. Experimental results show that TiO2 increased DLC bactericidal activity. Pure DLC films were thermodynamically unfavorable to bacterial adhesion. However, the chemical interaction between the E. coli and the studied films increased for the films with higher TiO2 concentration. As TiO2 bactericidal activity starts its action by oxidative damage to the bacteria wall, a decrease in the interfacial energy of bacteria adhesion causes an increase in the chemical interaction between E. coli and the films, which is an additional factor for the increasing bactericidal activity. From these results, DLC with TiO2 nanoparticles can be useful for producing coatings with antibacterial properties.

Photoacoustic monitoring of the absorption of isotonic saline solution by human mucus.

Viscosity and elasticity are the fundamental rheologic properties of respiratory mucus, and are important determinants of transportability of mucus in the mucociliary system. One technique that permits to monitor indirectly the rheologic properties of any sample is the photoacoustic technique. Using that technique, the absorption of isotonic saline solution by human mucus was monitored. The mucus was obtained from 11 volunteers, divided into two groups: five volunteers presenting pneumology symptoms (group I) and six healthy volunteers (group II). The photoacoustic signal of the mucus absorbing the saline solution was monitored as function of time, with measurements being performed each 10 min, up to 120 min. The resulting curves were fitted to sigmoidal curves to simulate the evolution on time of the photoacoustic signal. A characteristic time for the half saturation of the absorption process was obtained. For group I the time obtained was 23.3+/-5.3 min and for group II the time obtained was 55.0+/-7.7 min, both means being significantly different (Student t-test, p<0.05). This result supports the empirical practice of treating individuals presenting symptoms of airway obstruction with about 30 min of inhalations of isotonic saline solution vapor for the clearance of the airways.