Biocompatible and bioactive nanostructured glass coatings synthesized by pulsed laser deposition: In vitro biological tests

We report on the synthesis by pulsed laser deposition with a KrF* excimer laser source (λ = 248 nm, τ = 25 ns) of bioglass thin films of 6P57 and 6P61 types. Physiology, viability, and proliferation of human osteoblast cells were determined by quantitative in vitro tests performed by flow cytometry on primary osteoblasts cultured on pulsed laser deposited bioglasses. Both types of glass films proved to be appropriate mediums for cell survival and proliferation. In a parallel investigation, cell morphology and adhesion to the surface was studied by fluorescence microscopy and scanning electron microscopy. Strong bonds between the materials and cells were found in both cases, as osteoblast pseudopodes penetrated deep into the material. According to our observations, the 6P57 glass films were superior with respect to viability and proliferation performances.     

Surface characteristics and in vitro biocompatibility of a manganese-containing titanium oxide surface

This study investigated the surface characteristics and in vitro biocompatibility of a titanium (Ti) oxide layer incorporating the manganese ions (Mn) obtained by hydrothermal treatment with the expectation of utilizing potent integrin-ligand binding enhancement effect of Mn for future applications as an endosseous implant surface. The surface characteristics were evaluated by scanning electron microscopy, thin-film X-ray diffractometry, X-ray photoelectron spectroscopy, optical profilometry and inductively coupled plasma-atomic emission spectroscopy (ICP-AES). The in vitro biocompatibility of the Mn-containing Ti oxide surface was evaluated in comparison with untreated bare Ti using a mouse calvaria-derived osteoblastic cell line (MC3T3-E1). The hydrothermal treatment produced a nanostructured Mn-incorporated Ti oxide layer approximately 0.6 μm thick. ICP-AES analysis demonstrated that the Mn ions were released from the hydrothermally treated surface into the solution. Mn incorporation notably decreased cellular attachment, spreading, proliferation, alkaline phosphatase activity, and osteoblast phenotype gene expression compared with the bare Ti surface (p < 0.05). The results indicate that the Mn-incorporation into the surface Ti oxide layer has no evident beneficial effects on osteoblastic cell function, but instead, actually impaired cell behavior.     

Influence of surface topography and pore architecture of alkali-treated titanium on in vitro apatite deposition

Alkali-treated titanium surfaces have earlier shown to induce bone-like apatite deposition. In the present study, the effect of surface topography of two-dimensional and pore architecture of three-dimensional alkali-treated titanium substrates on the in vitro bioactivity was investigated. Titanium plates with a surface roughness of R_a = 0.13 μm, 0.56 μm, 0.83 μm, and 3.63 μm were prepared by Al₂O₃ grit-blasting. Simple tetragonal and face-centered Ti6Al4V scaffolds with spatial gaps of 450-1100 μm and 200-700 μm, respectively, were fabricated by a three-dimensional fiber deposition (3DFD) technique. After alkali treatment, the titanium plates with a surface roughness of R_a = 0.56 μm were completely covered with hydroxyapatite globules after 7 days in simulated body fluid (SBF), while the coverage of the samples with other surface roughness values remained incomplete. Similarly, face-centered Ti₆Al₄ scaffolds with spatial gaps of 200-700 μm exhibited a full surface coverage after 21 days in SBF, while simple tetragonal scaffolds with spatial gaps of 450-1100 μm were only covered for 45-65%. This indicates the importance of surface topography and pore architecture for in vitro bioactivity.     

Characterization on titanium surfaces and its effect on photocatalytic bactericidal activity

Many studies have been published on the use of TiO₂ as a photocatalyst, which decomposes various organic compounds under UV illumination by generating various radicals. The purpose of the present study was to evaluate the photocatalytic bactericidal effects of variously treated titanium surfaces on Escherichia coli K-12. The specimens were fabricated from grade 4 commercially pure titanium, 12 mm in diameter and 1 mm in thickness. Five different surfaces were prepared (MA: machined surface; AO: anodized at 300 V; NO: NaOH-treated; NW: NaOH- and water-treated; and HT: heat-treated). Surface analysis was performed using scanning electron microscopy, optical interferometer, and thin-film X-ray diffractometry. Photocatalytic activity of each group was confirmed by degradation of methylene blue (MB). The antibacterial activity was assessed by calculating the survival ratio in a drop of a culture of E. coli placed on the surface under UV illumination. Significant photocatalytic activity and bactericidal effects were observed on the titanium surfaces of AO and NW, regardless of the surface roughness (P < 0.01). The group with anatase was the most susceptible to the photocatalytic effect, while the surface without anatase showed the least susceptibility. Based on this in vitro study, the crystallography of the oxide layer on its titanium surfaces is an important factor affecting the photocatalytic bactericidal activity.     

Zinc-ion implanted and deposited titanium surfaces reduce adhesion of Streptococccus mutans

While titanium (Ti) is a commonly used dental implant material with advantageous biocompatible and mechanical properties, native Ti surfaces do not have the ability to prevent bacterial colonization. The objective of this study was to evaluate the chemical composition and bacterial adhesive properties of zinc (Zn) ion implanted and deposited Ti surfaces (Zn-PIIID-Ti) as potential dental implant materials. Surfaces of pure Ti (cp-Ti) were modified with increasing concentrations of Zn using plasma immersion ion implantation and deposition (PIIID), and elemental surface compositions were characterized by X-ray photoelectron spectrometry (XPS). To evaluate bacterial responses, Streptococcus mutans were seeded onto the modifiedTi surfaces for 48 h and subsequently observed by scanning electron microscopy. Relative numbers of bacteria on each surface were assessed by collecting the adhered bacteria, reculturing and counting colony forming units after 48 h on bacterial grade plates. Ti, oxygen and carbon elements were detected on all surfaces by XPS. Increased Zn signals were detected on Zn-PIIID-Ti surfaces, correlating with an increase of Zn-deposition time. Substantial numbers of S. mutans adhered to cp-Ti samples, whereas bacterial adhesion on Zn-PIIID-Ti surfaces signficantly decreased as the Zn concentration increased (p < 0.01). In conclusion, PIIID can successfully introduce Zn onto a Ti surface, forming a modified surface layer bearing Zn ions that consequently deter adhesion of S. mutans, a common bacterium in the oral environment.     

Surface structures and osteoblast response of hydrothermally produced CaTiO3 thin film on Ti-13Nb-13Zr alloy

This study investigated the surface characteristics and in vitro biocompatibility of a titanium (Ti) oxide layer incorporating calcium ions (Ca) obtained by hydrothermal treatment with or without post heat-treatment in the Ti-13Nb-13Zr alloy. The surface characteristics were evaluated by scanning electron microscopy, thin-film X-ray diffractometry, X-ray photoelectron spectroscopy, atomic force microscopy and contact angle measurements. In vitro biocompatibility of the Ca-containing surfaces was assessed in comparison with untreated surfaces using a pre-osteoblast cell line. Hydrothermal treatment produced a crystalline CaTiO₃ layer. Post heat-treatment at 400 °C for 2 h in air significantly decreased water contact angles in the CaTiO₃ layer (p < 0.001). The Ca-incorporated alloy surfaces displayed markedly increased cell viability and ALP activity compared with untreated surfaces (p < 0.001), and also an upregulated expression of various integrin genes (α1, α2, α5, αv, β1 and β3) at an early incubation time-point. Post heat-treatment further increased attachment and ALP activity in cells grown on Ca-incorporated Ti-13Nb-13Zr alloy surfaces. The results indicate that the Ca-incorporated oxide layer produced by hydrothermal treatment and a simple post heat-treatment may be effective in improving bone healing in Ti-13Nb-13Zr alloy implants by enhancing the viability and differentiation of osteoblastic cells.     

Immunocytochemical and structural comparative study of committed versus multipotent stem cells cultured with different biomaterials

The aim of this work was the comparison of the behavior of committed (human osteoblast cells – hOB – from bone biopsies) versus multipotent (human dental pulp stem cells – hDPSC – from extracted teeth) cells, cultured on shot-peened titanium surfaces, since the kind of cell model considered has been shown to be relevant in techniques widely used in studies on composition/morphology of biomaterial surfaces. The titanium surface morphology, with different roughness, and the behavior of cells were analyzed by confocal microscope (CM), scanning electron microscope (SEM) and X-ray microanalysis. The best results, in terms of hOB adhesion/distribution, were highlighted by both CM and SEM in cultured plates having 20-μm-depth cavities. On the contrary, CM and SEM results highlighted the hDPSC growth regardless the different surface morphology, arranged in overlapped layers due to their high proliferation rate, showing their unfitness in biomaterial surface test. Nevertheless, hDPSC cultured inside 3D-matrices reproduced an osteocyte-like three-dimensional network, potentially useful in the repair of critical size bone defects. The behavior of the two cell models suggests a different use in biomaterial cell cultures: committed osteoblast cells could be appropriate in selecting the best surfaces to improve osseointegration, while multipotent cells could be suitable to obtain in vitro osteocyte-like network for regenerative medicine. The originality of the present work consists in studying for the first time two different cell models (committed versus multipotent) compared in parallel different biomaterial cultures, thus suggesting distinct targets for each cellular model.     

Enhanced cell affinity of poly(l-lactide) film by immobilizing phosphonized chitosan

Graft polymerization of acrylic acid (AA) onto poly(l-lactide) (PLLA) film by UV irradiation was carried out to develop surfaces for N-methylene phosphonic chitosan (NMPC) immobilization. The properties of modified films were discussed by colorimetric method, attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR), contact angles, atomic force microscopy (AFM) and osteoblast incubation. The results showed that AA solution concentration and irradiation time had effect on the graft carboxyl densities. Comparing the ATR-FTIR images, two new peaks at 1561 cm⁻¹ and 1632 cm⁻¹ proved that NMPC was immobilized on the film surface successfully. The water contact-angles were decreased from 90 ± 5° to 37 ± 5° after modification. The AFM images indicated that the surface of the combined film was rougher than that of untreated film. The grafted film provided an excellent substrate for the growth of osteoblast.     

Preparation and in vitro characterization of aerosol-deposited hydroxyapatite coatings with different surface roughnesses

Hydroxyapatite (HA) coatings with different surface roughnesses were deposited on a Ti substrate via aerosol deposition (AD). The effect of the surface roughness on the cellular response to the coating was investigated. The surface roughness was controlled by manipulating the particle size distribution of the raw powder used for deposition and by varying the coating thickness. The coatings obtained from the 1100 °C-heated powder exhibited relatively smooth surfaces, whereas those fabricated using the 1050 °C-heated powder had network-structured rough surfaces with large surface areas and were superior in terms of their adhesion strengths and in vitro cell responses. The surface roughness (R_a) values of the coatings fabricated using the 1050 °C-heated powder increased from approximately 0.65 to 1.03 μm as the coating thickness increased to 10 μm. The coatings with a rough surface had good adhesion to the Ti substrate, exhibiting high adhesion strengths ranging from 37.6 to 29.5 MPa, depending on the coating thickness. The optimum biological performance was observed for the 5 μm-thick HA coating with an intermediate surface roughness value of 0.82 μm.     

Modified titanium surface with gelatin nano gold composite increases osteoblast cell biocompatibility

This study examined the gelatin nano gold (GnG) composite for surface modification of titanium in addition to insure biocompatibility on dental implants or biomaterials. The GnG composite was constructed by gelatin and hydrogen tetrachloroaurate in presence of reducing agent, sodium borohydrate (NabH₄). The GnG composite was confirmed by UV-VIS spectroscopy and transmission electron microscopy (TEM). A dipping method was used to modify the titanium surface by GnG composite. Surface was characterized by scanning electron microscopy (SEM) and energy dispersive X-ray (EDX). The MC-3T3 E1 cell viability was assessed by trypan blue and the expression of proteins to biocompatibility were analyzed by Western blotting. The GnG composite showed well dispersed character, the strong absorption at 530 nm, roughness, regular crystal and clear C, Na, Cl, P, and Au signals onto titanium. Further, this composite allowed MC-3T3 E1 growth and viability compared to gelatin and pure titanium. It induced ERK activation and the expression of cell adherent molecules, FAK and SPARC, and growth factor, VEGF. However, GnG decreased the level of SAPK/JNK. This shows that GnG composite coated titanium surfaces have a good biocompatibility for osteoblast growth and attachment than in intact by simple and versatile dipping method. Furthermore, it offers good communication between cell and implant surfaces by regulating cell signaling and adherent molecules, which are useful to enhance the biocompatibility of titanium surfaces.     

Comparative assessment of optical performance for the optical design of HD-DVD micro objective lens with NA 0.65 using aspheric and freeform surfaces

A micro objective lens for HD-DVD with the Numerical Aperture (NA) of 0.65 at blue laser wavelength with clear aperture diameter of 1.5 mm has been designed on acrylic material using freeform surfacing method. Optical performance evaluation parameters have been compared with bi-aspheric surfaces based objective lens design. Freeform surface based design has higher degrees of freedom compare to conventional aspheric surfaces that can reduce aberrations significantly. Maximum RMS error is 0.003λ at 0.4° and maximum RMS radius is 0.027 μm while airy disk radius is 0.3803 μm for freeform surface based design. Hence single freeform surface based design can reduce the fabrication complexity and tooling time, at the same instance provides comparable performance with bi-aspheric surfaces based design.     

Surface microstructures and antimicrobial properties of copper plasma alloyed stainless steel

Bacterial adhesion to stainless steel surfaces is one of the major reason causing the cross-contamination and infection in many practical applications. An approach to solve this problem is to enhance the antibacterial properties on the surface of stainless steel. In this paper, novel antibacterial stainless steel surfaces with different copper content have been prepared by a plasma surface alloying technique at various gas pressures. The microstructure of the alloyed surfaces was investigated using glow discharge optical emission spectroscopy (GDOES) and scanning electron microscopy (SEM). The viability of bacteria attached to the antibacterial surfaces was tested using the spread plate method. The antibacterial mechanism of the alloyed surfaces was studied by X-ray photoelectron spectroscopy (XPS). The results indicate that gas pressure has a great influence on the surface elements concentration and the depth of the alloyed layer. The maximum copper concentration in the alloyed surface obtained at the gas pressure of 60 Pa is about 7.1 wt.%. This alloyed surface exhibited very strong antibacterial ability, and an effective reduction of 98% of Escherichia coli (E. coli) within 1 h was achieved by contact with the alloyed surface. The maximum thickness of the copper alloyed layer obtained at 45 Pa is about 6.5 μm. Although the rate of reduction for E. coli of this alloyed surface was slower than that of the alloyed surface with the copper content about 7.1 wt.% over the first 3 h, few were able to survive more than 12 h and the reduction reached over 99.9%. The XPS analysis results indicated that the copper ions were released when the copper alloyed stainless steel in contact with bacterial solution, which is an important factor for killing bacteria. Based on an overall consideration of bacterial killing rate and durability, the alloyed surface with the copper content of 2.5 wt.% and the thickness of about 6.5 μm obtained at the gas pressure of 45 Pa is expected to be useful as antimicrobial materials that may have a promising future in antimicrobial applications.     

Preparation of superhydrophobic films on titanium as effective corrosion barriers

Stable superhydrophobic films were prepared on the electrochemical oxidized titania/titanium substrate by a simple immersion technique into a methanol solution of hydrolyzed 1H,1H,2H,2H-perfluorooctyltriethoxysilane [CF₃(CF₂)₅(CH₂)₂Si(OCH₂CH₃)₃, PTES] for 1 h at room temperature followed by a short annealing at 140 °C in air for 1 h. The surface morphologies and chemical composition of the film were characterized by means of water contact angle (CA), field emission scanning electron microscopy (FESEM), atomic force microscope (AFM) and X-ray photoelectron spectroscopy (XPS). The water contact angle on the surface of this film was measured to be as high as 160°. SEM images showed that the resulting surfaces exhibited special hierarchical structure. The special hierarchical structure along with the low surface energy leads to the high surface superhydrophobicity. The corrosion resistance ability and durance property of the superhydrophobic film in 3.5 wt.% NaCl solution was evaluated by the electrochemical impedance spectroscopy (EIS). The anticorrosion properties of the superhydrophobic film are compared to those of unmodified pure titanium and titania/titanium substrates. The results showed that the superhydrophobic film provides an effective corrosion resistant coating for the titanium metal even with immersion periods up to 90 d in the 3.5 wt.% NaCl solution, pointing to promising future applications.     

Laser shock wave consolidation of nanodiamond powders on aluminum 319

A novel coating approach, based on laser shock wave generation, was employed to induce compressive pressures up to 5 GPa and compact nanodiamond (ND) powders (4-8 nm) on aluminum 319 substrate. Raman scattering indicated that the coating consisted of amorphous carbon and nanocrystalline graphite with peaks at 1360 cm⁻¹ and 1600 cm⁻¹ respectively. Scanning electron microscopy revealed a wavy, non-uniform coating with an average thickness of 40 μm and absence of thermal effect on the surrounding material. The phase transition from nanodiamond to other phases of carbon is responsible for the increased coating thickness. Vicker's microhardness test showed hardness in excess of 1000 kg_f/mm² (10 GPa) while nanoindentation test indicated much lower hardness in the range of 20 MPa to 2 GPa. Optical surface profilometry traces displayed slightly uneven surfaces compared to the bare aluminum with an average surface roughness (R_a) in the range of 1.5-4 μm depending on the shock wave pressure and type of confining medium. Ball-on-disc tribometer tests showed that the coefficient of friction and wear rate were substantially lower than the smoother, bare aluminum sample. Laser shock wave process has thus aided in the generation of a strong, wear resistant, durable carbon composite coating on aluminum 319 substrate.     

Scattering from unidirectional ground steel surfaces in the specular direction

Scattering in the specular direction from unidirectional ground steel surfaces having random roughness is studied theoretically and experimentally. The grooves were oriented perpendicular and parallel to the plane of incidence and were illuminated by a light beam that was smaller than the sample size. Expressions for scattering from a one-dimensional rough surface in the specular direction were derived for the both orientations of the grooves. For the same groove orientations, scattering (λ = 0.633 μm) from the ground surfaces was measured in the specular direction at angles of incidence from 6° to 82°. The measured rms roughnesses of the surfaces were 0.096 μm, 0.143 μm, 0.311 μm, and 0.501 μm, respectively. The measured scattering was independent of the orientation of the grooves for the two smoother surfaces and depended on the groove orientation for the other surfaces. Calculations using the derived expressions taking into account the experimental results show that the scattering is independent of the orientation of the grooves if the rms roughness is no larger than ∼0.16 μm.     

Cell adhesive ability of a biological foam ceramic with surface modification

Biological foam ceramic is a promising material for tissue engineering scaffold because of its biocompatibility, biodegradation and adequate pores measured from micrometer to nanometers. The aim of this study was to evaluate the adhesion and proliferation of adipose-derived stromal cells (ADSCs) on the biological foam ceramic coated with fibronectin. ADSCs were harvested from SD rats and passaged three times prior to seeding onto biological foam surface modified with fibronectin (50 μg/ml). Scaffold without surface modification served as control. To characterize cellular attachment, cells were incubated on the scaffold for 1 h and 3 h and then the cells attached onto the scaffold were counted. The difference of proliferation was appraised using MTT assay at day 1, 3, 5 and 7 before the cells reached confluence. After 7 days of culture, scanning electron microscope (SEM) was chosen to assess cell morphology and attachment of ADSCs on the biological foam ceramic. Attachment of ADSCs on the biological foam ceramic surface modified with fibronectin at 1 h or 3 h was substantially greater than that in control. MTT assay revealed that ADSCs proliferation tendency of the experimental group was nearly parallel to that of control. SEM view showed that ADSCs in the experimental groups connected more tightly and excreted more collagen than that in control. The coating of fibronectin could improve the cell adhesive ability of biological foam ceramics without evident effect on proliferation.     

A cross-sectional scanning tunneling microscopy study of IrO2 rutile single crystals

Rutile iridium dioxide (IrO₂) surfaces were studied by cross-sectional scanning tunneling microscopy (XSTM). Atomically flat surfaces prepared by in situ ultra high vacuum cleaving of crystalline platelets of thicknesses <50 μm were successfully demonstrated. In addition to (1 1 0) surface, several vicinal planes, e.g., (1 2 0), (1 3 0) and , were also examined. The cleaved planes are close to bulk-terminated surfaces with predominant [0 0 1]-oriented bridge oxygen rows. Unlike TiO₂, bright oxygen rows are imaged and oxygen defects appear as dim species. Our studies show that XSTM is a viable technique to study oxide surfaces that are otherwise difficult to prepare.     

Ultraslow phonon-assisted collapse of tubular image states

We predict ultraslow collapse of “tubular image states” (TIS) on material surfaces. TIS are bound Rydberg-like electronic states formed at large distances (∼30 nm) from the surfaces of suspended circularly-symmetric nanowires, such as metallic C nanotubes. The states are formed in potential wells, resulting from a combination of the TIS-electron attraction to image charges in the nanotube and its centrifugal repulsion, caused by spinning around the tube. We demonstrate that TIS can collapse on the tube surface by passing their angular momentum l to circularly polarized flexural phonons excited in the tube. Our analysis shows that for highly detached TIS with l ? 6 the relaxation lifetimes are of the order of 10 ns-1 μs, while for l < 6 these lifetimes are reduced by several orders of magnitude.     

Surface modification of polyethylene terephthalate with albumin and gelatin for improvement of anticoagulation and endothelialization

The surfaces of polyethylene terephthalate (PET) were modified by oxygen plasma-induced and ultraviolet (UV)-assisted acrylic acid (AAc) grafting polymerization, and the carboxyl (COOH) groups on the PET surface was 5.29 × 10⁻⁹mol/cm². Then using the COOH as reacting sites, the molecules of gelatin and bovine serum albumin (BSA) were further co-immobilized on the PET surface. The modified PET surfaces were characterized by X-ray photoelectron spectroscopy (XPS) and surface chemical quantitative analysis. The results showed that the molecules of gelatin and albumin were immobilized on the PET surface. The concentration of gelatin on the gelatin-immobilized PET surface was 2.02 μg/cm². For the gelatin-immobilized PET surface, the human umbilical vein endothelial cells (HUVECs) culture attachment and proliferation ratios were improved, but the anticoagulation became worse proved by platelet adhesion test in vitro and the lactate dehydrogense (LDH) test. After further co-immobilization of albumin with gelatin biomolecules on the PET surface (PET-Gel-BSA), the percent of platelet adhesion in vitro decreased 28% than that on the gelatin-immobilized PET surface, and the cell density on the PET-Gel-BSA film (1.08 × 10⁵ cells/cm²) was significantly higher than that on the control PET surface. This investigation tries to find a method which can construct the anticoagulant surface before the endothelium formation and also accelerate the endothelialization of polymer surface.