In-vitro bioactivity,biocorrosion and antibacterial activity of silicon integrated hydroxyapatite/chitosan composite coating on 316 L stainless steel implants

A simple and effective ultrasonication method was applied for the preparation of 0, 0.4, 0.8, 1.0 and 1.6 wt% silicon substituted hydroxyapatite (HAp) (SH). The Ca/P ratio of the synthesised SH nanoparticles were in the range of 1.58–1.70. Morphological changes were noticed in HAp with respect to the amount of Si from 0 to 1.6 wt%. The morphology of the particles changed from spherical shape to rod-like morphology with respect to the amount of Si which was confirmed using transmission electron microscopy. X-ray diffraction studies confirm the formation of phase pure SH nanoparticles without any secondary phase. Chitosan (CTS) blended SH nanocomposites coating on surgical grade 316 L stainless steel (316 L SS) implant was made by spin coating technique. The surface of the coated implant was characterised using scanning electron microscopy which confirms the uniform coating without cracks and pores. The increased corrosion resistance of the 1.6 wt% of SH/CTS-coated SS implant in the simulated body fluid (SBF) indicates the long-term biostability of SH composite-coated ceramics in vitro than the 0 wt% SH/CTS. The testing of SH/CTS nanocomposites with gram-positive and gram-negative bacterial strains confirms that the antibacterial ability improves with the higher substitution of Si. In addition, formation of bone-like apatite layer on the SH/CTS-coated implant in SBF was studied through SEM analysis and it confirms the ability to increase the HAp formation on the surface of 1.0 wt% SH/CTS-coated 316 L SS implant.     

An investigation of the biochemical properties of tetrazines as potential coating additives

1,2,4,5-Tetrazine and its 3,6-disubstituted derivatives are currently used for a range of industrial and medical applications as they exhibit particular coordination chemistries, characterised by electron and charge transfer phenomena. The aim of the present work is to synthesise two tetrazine derivatives, namely 3,6-dihydrazino-1,2,4,5-tetrazine (DHDTZ) and 1,2,4,5-tetrazine dicarboxylic acid (DCTZ), and determine their antibacterial, antioxidant and anticorrosion characteristics as additives in a sol–gel coating on SS316L steel. The structure of the tetrazines was confirmed by NMR and FTIR while the surface morphology of bacterial cells in their presence was observed by AFM. Their ability to inhibit corrosion on 316L stainless steel was electrochemically determined using a potentiodynamic scanning (PDS) technique. The corrosion inhibition results showed that the acidic DCTZ provided the best corrosion protection. The concentration-dependent antioxidant capacity of the tetrazines was confirmed by both DPPH radical scavenging activity and FRAP assays, showing higher activity for DHDTZ than DCTZ. Furthermore, a DHDTZ doped sol–gel solution was prepared and curing parameter (temperature and time) was optimised for coating on microtitre wells and stainless steel panel. The antibacterial activity of the coated surfaces against Pseudomonas aeruginosa ATCC 27853 and the biofilm forming bacteria Staphylococcus epidermidis CSF 41498 was determined. DHDTZ showed significantly higher antibacterial activities with MIC as low as 31 ppm compared to 250 ppm for DCTZ.     

Annealed thin-film zirconia coating adhered on 316L stainless steel as a bio-inert indwelling needle

A 316L stainless steel plate was covered with a ZrO₂ coating using the sol–gel dip coating technique, and preliminary in vitro and in vivo studies were conducted. The morphology and crystal structure of the coatings were examined using scanning electron microscope and X-ray diffraction, and their surface chemical structures were characterised using X-ray photoelectron spectroscopy. The quality and adhesion of the coatings on the substrate were measured using a nano-indenter with lateral force and scratch modes. ZrO₂ was formed on a 316L plate at various temperatures, resulting in different crystalline structures, surface morphologies, and integration with the 316L surface. The optimal conditions to produce ZrO₂–316L are an annealing temperature of 500 °C and duration of 1 h (ZrO₂/316L_500), yielding an adhesive force of 595 μN. 3T3 cell morphology, adhesion, and viability using the live/dead cell staining protocol were assessed. Cell affinity was significantly enhanced on the surface of ZrO₂/316L_500, compared to the as prepared sample. Furthermore, after mice were injected with 316L and ZrO₂/316L_500 needles for durations of up to 72 h, wound contraction, inflammation, and proliferation were compared. The results indicate that the ZrO₂/316L_500 needle exhibits high potential as a bio-inert coating and that it can be applied to scalpels and indwelling needles.     

Novel hydroxyapatite/tantalum surface coating for metallic dental implant

The aim of this study was to design and produce a novel surface composite coating on metallic substrate in order to improve the biocompatibility of metallic dental implant and the bone osteointegration simultaneously.Stainless steel 316L (SS) was used as a metallic substrate and a novel double-layer hydroxyapatite/tantalum (HA/Ta) coating was prepared on it. Tantalum coating was made using physical vapor deposition process and HA coating was produced using plasma-spraying technique on it. X-ray diffraction (XRD) and scanning electron microscopy (SEM) techniques were utilized to investigate the coating characterization. Electrochemical polarization tests were performed in two types of physiological solutions at 37 ± 1 °C in order to determine the corrosion behavior of the coated and uncoated specimens as indication of biocompatibility.The results indicated that the decrease in corrosion current density was significant for HA/Ta coated specimens and was much lower than the value obtained for uncoated 316L SS. The novel double-layer HA/Ta composite coating could improve the corrosion resistance and thus the biocompatibility of 316L SS dental implant.     

Quantification of the adhesion free energy between bacteria and hydrophobic and hydrophilic substrata

In this paper, the bacterial adhesion to hydrophilic glass and hydrophobic indium tin oxide (ITO) coated glass surfaces was investigated. The aim of this work was to examine the effect of the hydrophobicity of two bacterial strains (Pseudomonas stutzeri PS, and Staphylococcus epidermidis SE) and substrata surfaces on the adhesion mechanism. The hydrophobicity of the surfaces of the bacterial strains as well as of the surfaces of the tested materials was characterized using the contact angle measurement via sessile drop technique. The water contact angle measurements of the intrinsic cell hydrophobicity showed that the PS strain is more hydrophobic (θ = 40°) than SE strain (θ = 21°). For both tested materials, this angle was 12° for glass surface and 84° for ITO-coated glass surface. Based on the thermodynamic approach, the Lifshitz van der Waals interaction free energy, the hydrophobic interaction free energy and the total interaction free energy between the bacteria and the substratum through the suspending medium were quantified. In order to verify the thermodynamic approach predictions of the bacterial adhesion on each substratum, adhesion tests were carried out for each bacteria/substratum combination. The results revealed that for both materials surfaces, the adhesion energy of the hydrophobic PS strain (26.1 mJ/m² for glass and − 3.8 mJ/m² for ITO-coated glass) is higher than that of SE strain (31.8 mJ/m² for glass and 14.3 mJ/m² for ITO-coated glass). For both bacterial strains, the effect of the hydrophobicity property of the substratum seems more important in the initial adhesion step. However, the second step of adhesion involves the biological approach, since a discrepancy was found between physicochemical theoretical approach and adhesion tests for SE strain.     

Localized corrosion behavior of 316L stainless steel in the presence of sulfate-reducing and iron-oxidizing bacteria

Localized corrosion behavior of 316L stainless steel was investigated in the presence of sulfate-reducing bacteria (SRB) and iron-oxidizing bacteria (IOB) isolated from cooling water system using polarization measurement, scanning electron microscopy (SEM) examinations and energy dispersive spectrum (EDS) analysis. The results show the corrosion potential (E_corr) and breakdown potential (E_b) of SS decreased in turn with the presence of IOB, SRB and SRB + IOB, indicating decreased relative resistance to localized corrosion. E_corr in the sterile medium remained virtually unchanged with exposure time, indicating that localized attack did not occur. However, micrometer-scale pitting was observed on the SS surface in the presence of bacteria. The presence of SRB demonstrated higher corrosion rates than IOB. The combination of SRB and IOB yielded the highest corrosion rate. The presence and metabolic activities of bacteria on SS surface produce environments that can alter rates of partial reactions in corrosion processes and shift corrosion mechanisms. The most severe microbiologically induced corrosion takes place in aquatic solution where physiological groups of aerobic and anaerobic microorganisms interact.     

Studies of calcium-precipitating oral bacterial adhesion on TiN,TiO2 single layer,and TiN/TiO2 multilayer-coated 316L SS

Titanium nitride (TiN), titanium oxide (TiO₂) single layer, and TiN/TiO₂ multilayer coatings were deposited on a 316L stainless steel substrate using reactive magnetron sputtering process with the aim of preventing bacterial adhesion. The crystal structures of as-prepared coatings were evaluated using X-ray diffraction analysis. The cubic structure of TiN, anatase, and rutile structure of TiO₂ was noticed. Atomic force microscopy images exhibited a relatively smooth surface for all coatings. The surface wettability studies confirmed that the coatings were hydrophilic in nature. The rate of bacterial adhesion was evaluated using scanning electron microscopy and epifluorescence microscopy. These results demonstrated that the coated substrates could help to effectively reduce the bacterial adhesion and biofilm formations.     

Effect of the La alloying addition on the antibacterial capability of 316L stainless steel

316L stainless steel is widely used for fashion jewelry but it can carry a large number of bacteria and cause the potential risk of infection since it has no antimicrobial ability. In this paper, La is used as an alloying addition. The antibacterial capability, corrosion resistance and processability of the La-modified 316L are investigated by microscopic observation, thin-film adhering quantitative bacteriostasis, electrochemical measurement and mechanical test. The investigations reveal that the La-containing 316L exhibits the Hormesis effect against Staphylococcus aureus ATCC 25923 and Escherichia coli DH5α, 0.05 wt.% La stimulates their growth, as La increases, the modified 316L exhibits the improved antibacterial effect. The more amount of La is added, the better antibacterial ability is achieved, and 0.42 wt.% La shows excellent antibacterial efficacy. No more than 0.11 wt.% La addition improves slightly the corrosion resistance in artificial sweat and has no observable impact on the processability of 316L, while a larger La content degrades them. Therefore, the addition of La alone in 316L is difficult to obtain the optimal combination of corrosion resistance, antibacterial capability and processability. In spite of that, 0.15 wt.% La around is inferred to be the trade-off for the best overall performance.     

Colloidal drug probe: Method development and validation for adhesion force measurement using Atomic Force Microscopy

This study demonstrates a novel technique of preparing drug colloid probes to determine the adhesion force between a model drug salbutamol sulphate (SS) and the surfaces of polymer microparticles to be used as carriers for the dispersion of drug particles from dry powder inhaler (DPI) formulations. Model silica probes of approximately 4 μm size, similar to a drug particle used in DPI formulations, were coated with a saturated SS solution with the aid of capillary forces acting between the silica probe and the drug solution. The developed method of ensuring a smooth and uniform layer of SS on the silica probe was validated using X-ray Photoelectron Spectroscopy (XPS) and Scanning Electron Microscopy (SEM). Using the same technique, silica microspheres pre-attached on the AFM cantilever were coated with SS. The adhesion forces between the silica probe and drug coated silica (drug probe) and polymer surfaces (hydrophilic and hydrophobic) were determined. Our experimental results showed that the technique for preparing the drug probe was robust and can be used to determine the adhesion force between hydrophilic/hydrophobic drug probe and carrier surfaces to gain a better understanding on drug carrier adhesion forces in DPI formulations.     

Oxygen plasma functionalization of parylene C coating for implants surface: Nanotopography and active sites for drug anchoring

The effect of oxygen plasma treatment (t = 0.1–60 min, p_O2 = 0.2 mbar, P = 50 W) of parylene C implant surface coating was investigated in order to check its influence on morphology (SEM, AFM observations), chemical composition (XPS analysis), hydrophilicity (contact angle measurements) and biocompatibility (MG-63 cell line and Staphylococcus aureus 24167 DSM adhesion screening). The modification procedure leads to oxygen insertion (up to 20 at.%) into the polymer matrix and together with surface topography changes has a dramatic impact on wettability (change of contact angle from θ = 78 ± 2 to θ = 33 ± 1.9 for unmodified and 60 min treated sample, respectively). As a result, the hydrophilic surface of modified parylene C promotes MG-63 cells growth and at the same time does not influence S. aureus adhesion. The obtained results clearly show that the plasma treatment of parylene C surface provides suitable polar groups (C = O, C–O, O–C = O, C–O–O and O–C(O)–O) for further development of the coating functionality.     

Antibacterial effects of silver-doped hydroxyapatite thin films sputter deposited on titanium

Since many orthopedic implants fail as a result of loosening, wear, and inflammation caused by repeated loading on the joints, coatings such as hydroxyapatite (HAp) on titanium with a unique topography have been shown to improve the interface between the implant and the natural tissue. Another serious problem with long-term or ideally permanent implants is infection. It is important to prevent initial bacterial colonization as existing colonies have the potential to become encased in an extracellular matrix polymer (biofilm) that is resistant to antibacterial agents. In this study, plasma-based ion implantation was used to examine the effects of pre-etching on plain titanium. Topographical changes to the titanium samples were examined and compared via scanning electron microscopy. Hydroxyapatite and silver-doped hydroxyapatite thin films were then sputter deposited on titanium substrates etched at ? 700 eV. For silver-doped films, two concentrations of silver (~ 0.5 wt.% and ~ 1.5 wt.%) were used. Silver concentrations in the film were determined using energy dispersive X-ray spectroscopy. Hydroxyapatite film thicknesses were determined by measuring the surface profile using contact profilometry. Staphylococcus epidermidis and Pseudomonas aeruginosa adhesion studies were performed on plain titanium, titanium coated with hydroxyapatite, titanium coated with ~ 0.5 wt.% silver-doped hydroxyapatite, and titanium coated with ~ 1.5 wt.% silver-doped hydroxyapatite. Results indicate that less bacteria adhered to surfaces containing hydroxyapatite and silver; further, as the hydroxyapatite films delaminated, silver ions were released which killed bacteria in suspension.     

Cytocompatibility and mechanical properties of novel porous 316 L stainless steel

Novel 316 L stainless steel (SS) foam with 85% porosity and an open pore diameter of 70–440 μm was developed for hard tissue application. The foam sheet with a 200-μm diameter had superior cell proliferation and penetration as identified through in vitro experiments. Calcification of human osteosarcoma cells in the SS foam was observed. Multi-layered foam preparation is a potential alternative technique that satisfies multi-functional requirements such as cell penetration and binding strength to the solid metal. In tensile tests, Young's modulus and the strength of the SS foam were 4.0 GPa and 11.2 MPa respectively, which is comparable with human cancellous bone.     

Calcium phosphate coating containing silver shows high antibacterial activity and low cytotoxicity and inhibits bacterial adhesion

Surgical site infection is one of the serious complications of orthopedic implants. In order to reduce the incidence of implant-associated infections, we developed a novel coating technology of calcium phosphate (CP) containing silver (Ag), designated Ag-CP coating, using a thermal spraying technique. In this study, we evaluated the antibacterial efficacy and biological safety of this coating. In vitro antibacterial activity tests showed that the growths of Escherichia coli, Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) are completely suppressed on Ag-CP coating. In vitro bacterial adherence tests revealed that the number of adherent bacteria on the surface of this coating is significantly less (p < 0.02) than that on the surface of the CP coating. Moreover, the Ag-CP coating completely inhibits MRSA adhesion [<10 colony-forming units (CFU)] when 10² CFU MRSA is inoculated. On the other hand, V79 Chinese hamster lung cells were found to grow on the Ag-CP coating as well as on the CP coating in a cytotoxicity test. These results indicate that the Ag-CP coating on the surface of orthopedic implants exhibits antibacterial activity and inhibits bacterial adhesion without cytotoxicity.     

Effect of surface nanocrystallization induced by fast multiple rotation rolling on mechanical properties of a low carbon steel

Fast multiple rotation rolling (FMRR), a novel and efficient surface nanocrystallization technique, was used to fabricate a nanostructured layer in the surface of low carbon steel. The microstructure of the surface layer was characterized by transmission electron microscopy, optical microscope and scanning electron microscopy, and mechanical properties were investigated by microhardness measurements, tensile measurements and friction and wear tests. In addition, the fracture and wear scars morphologies were observed by scanning electron microscopy. Experimental results indicated that a deformation layer with thickness about 200 μm is clearly observed in the FMRR sample surface. A nanostructured layer of 30 μm thick is obtained, with grain size ranging from 8 to 18 nm and average grain size about 14 nm in the top surface layer. The microhardness of the FMRR sample change gradiently along the depth from about 316 HV in the top surface layer to about 160 HV in the matrix, which is nearly twice harder than that of the original sample. The ultimate tensile strength has also been markedly improved. And the friction and wear experiments show that tribological properties of the low carbon steel have been enhanced by FMRR treatment.     

Temperature responsive polymers as multiple function reagents in mineral processing

A processing scheme which uses a single chemical that has multiple functions to achieve both efficient mineral flotation and solids dewatering is presented. Temperature sensitive polymers which display hydrophilic/hydrophobic transitions in response to changes in temperature such as poly (N-isopropyl acrylamide) (PNIPAM) have been found to be useful as such multiple function reagents. This polymer can cause the mineral particles’ surfaces to be hydrophilic at temperature below the critical solution temperature (CST = 32 °C) and hydrophobic at temperature above the CST. Therefore, both particle surface wettability and inter-particle interaction forces are reversibly controllable. When the surface is hydrophilic, particle dispersion is achieved by repulsive inter-particle forces whereas when the surface is hydrophobic, particle aggregation is induced by inter-particle hydrophobic attractive forces. In addition, the hydrophobic surface condition allows for the attachment of particles to bubbles. Flotation and solid settling tests have been conducted with silica and kaolinite suspensions treated with (PNIPAM). Both effective suspension dispersion or hydrophobic aggregation and flotation without any additional collector have been demonstrated. In solid/liquid separation, rapid settling was obtained with hydrophobic aggregation at temperature above the CST and further sediment consolidation (and water release) occurred at temperature below the CST. The approach has the potential to reduce the amount and types of reagents required for mineral processing.     

Probing interaction of Gram-positive and Gram-negative bacterial cells with ZnO nanorods

In the present work, the physiological effects of the ZnO nanorods on the Gram positive (Staphylococcus aureus and Bacillus subtilis) and Gram-negative (Escherichia coli and Aerobacter aerogenes) bacterial cells have been studied. The analysis of bacterial growth curves for various concentrations of ZnO nanorods indicates that Gram positive and Gram negative bacterial cells show inhibition at concentrations of ~ 64 and ~ 256 μg/mL respectively. The marked difference in susceptibility towards nanorods was also validated by spread plate and disk diffusion methods. In addition, the scanning electron micrographs show a clear damage to the cells via changed morphology of the cells from rod to coccoid etc. The confocal optical microscopy images of these cells also demonstrate the reduction in live cell count in the presence of ZnO nanorods. These, results clearly indicate that the antibacterial activity of ZnO nanorods is higher towards Gram positive bacterium than Gram negative bacterium which indicates that the structure of the cell wall might play a major role in the interaction with nanostructured materials and shows high sensitivity to the particle concentration.     

In vitro antimicrobial properties of silver–polysaccharide coatings on porous fiber-reinforced composites for bone implants

Biostable fiber-reinforced composite (FRC) implants prepared from bisphenol-A-dimethacrylate and triethyleneglycoldimethacrylate resin reinforced with E-glass fibers have been successfully used in cranial reconstructions in 15 patients. Recently, porous FRC structures were suggested as potential implant materials. Compared with smooth surface, porous surface allows implant incorporation via bone ingrowth, but is also a subject to bacterial attachment. Non-cytotoxic silver–polysaccharide nanocomposite coatings may provide a way to decrease the risk of bacterial contamination of porous FRC structures. This study is focused on the in vitro characterization of the effect porosity on the antimicrobial efficiency of the coatings against Staphylococcus aureus and Pseudomonas aeruginosa by a series of microbiological tests (initial adhesion, antimicrobial efficacy, and biofilm formation). Characterization included confocal laser scanning microscopy and scanning electron microscopy. The effect of porosity on the initial attachment of S. aureus was pronounced, but in the case of P. aeruginosa the effect was negligible. There were no significant effects of the coatings on the initial bacterial attachment. In the antimicrobial efficacy test, the coatings were potent against both strains regardless of the sample morphology. In the biofilm tests, there were no clear effects either of morphology or of the coating. Further coating development is foreseen to achieve a longer-term antimicrobial effect to inhibiting bacterial implant colonization.     

Experimental and theoretical investigations of the adhesion time of Penicillium spores to cedar wood surface

In this study, the adhesion of 4 Penicillium strains (Penicillium granulatum, Penicillium crustosum, Penicillium commune and Penicillium chrysogenum) on cedar wood was examined qualitatively and quantitatively by using the extended DLVO (XDLVO) approach and the environmental scanning electronic microscopy (ESEM) technique. A comparison between the XDLVO theories and the ESEM technique was also investigated. The adhesion tests revealed that P. chrysogenum was not able to adhere on the cedar wood substrata, as predicted by the XDLVO approach. We have also found by ESEM that the three Penicillium strains (P. granulatum, P. crustosum, P. commune) adhered on wood, as not predicted theoretically.Moreover, the time of adhesion (3 h and 24 h) was used not only to compare the capacity of adhesion according to contact time but also to explain the discrepancies between the XDLVO approach prediction and the adhesion experiments. A positive relationship between the XDLVO approach and adhesion experiments has been observed after 3 h of adhesion. In contrast, a contradiction between the XDLVO predictions and the adhesion test results has been noted after 24 h of adhesion of Penicillium strains to the wood surface.     

Temperature-sensitivity and cell biocompatibility of freeze-dried nanocomposite hydrogels incorporated with biodegradable PHBV

The structure, morphology, thermal behaviors and cytotoxicity of novel hydrogels, composed of poly(N-isopropylacrylamide)(PNIPAM) and biodegradable polyester poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) under nanoclay hectorite “Laponite XLG” severed as physical cross-linker, were characterized by X-ray diffraction, scanning electron microscopy, gravimetric method, differential scanning calorimetry, and cell culture experiments. It was found that, due to the introduction of hydrophobic PHBV, the homogeneity of interior pore in the pure PNIPAM nanocomposite hydrogel was disrupted, the transparency and swelling degree gradually decreased. Although the weight ratio between PHBV and NIPAM increased from 5 to 40 wt.%, the volume phase transition temperature (VPTTs) of hydrogel were not altered compared with the pure PNIPAM nanocomposite hydrogel. No matter what PHBV content, the PHBV/PNIPAM/Hectorite hydrogels always exhibit good stimuli-responsibility. In addition, human hepatoma cells(HepG2) adhesion and spreading on the surface of PHBV-based hydrogels was greatly improved than that of pure PNIPAM nanocomposite hydrogel at 37 °C due to the introduction of PHBV.     

In vitro and in vivo antimicrobial properties of silver-containing hydroxyapatite prepared via ultrasonic spray pyrolysis route

Hydroxyapatite (HAp), with its high biocompatibility and osteoconductivity, readily absorbs proteins, amino acids and other substances, which in turn favor the adsorption and colonization of bacteria. To prevent bacterial growth and biofilm formation on HAp discs, silver-containing (1–20 mol%) HAp (Ag-HAp) powders were synthesized using an ultrasonic spray pyrolysis (USSP) technique. The X-ray diffraction (XRD) peaks were very broad, indicating low crystallinity, and this induced the release of Ag⁺ ions from Ag-HAp powders. In addition, a gradual increase in Ca^2 + ion release was observed. These results suggest that dissolution of Ca^2 + ion in Ag-HAp triggered the release of Ag⁺ ions.The antimicrobial efficacy of Ag-HAp disc was tested against Staphylococcus aureus. Samples with Ag contents of more than 5 mol% were found to be highly effective against bacterial colonization and biofilm formation in vitro. In vivo antibacterial tests using bioluminescent strains also showed reductions in the viability of bacteria with Ag-HAp (5 mol%) discs. Biocompatibility tests using a modified Transwell® insert method showed that Ag-HAp (5 mol%) discs have negative effects on osteoblast proliferation. These results indicate that Ag-HAp (5 mol%) has effective antibacterial activity and good biocompatibility both in vitro and in vivo together with good biocompatibility, thus confirming its utility as a bactericidal material.