Surface characteristics of dental implants: A review.

OBJECTIVES: During the last decades, several changes of paradigm have modified our view on how biomaterials' surface characteristics influence the bioresponse. After becoming aware of the role of a certain microroughness for improved cellular contact and osseointegration of dental titanium implants, the likewise important role of surface energy and wettability was increasingly strengthened. Very recently, synergistic effects of nanoscaled topographical features and hydrophilicity at the implant/bone interface have been reported. METHODS: Questions arise about which surface roughness and wetting data are capable to predict the bioresponse and, ultimately, the clinical performance. Current methods and approaches applied for topographical, wetting and surface energetic analyses are highlighted. Current knowledge of possible mechanisms explaining the influence of roughness and hydrophilicity at the biological interface is presented. RESULTS: Most marketed and experimental surfaces are based on commonly available additive or subtractive surface modifying methods such as blasting, etching or anodizing. Different height, spatial, hybrid and functional roughness parameters have been identified as possible candidates able to predict the outcome at hard and soft tissue interfaces. Likewise, hydrophilic implants have been proven to improve the initial blood contact, to support the wound healing and thereby accelerating the osseointegration. SIGNIFICANCE: There is clear relevance for the influence of topographical and wetting characteristics on a macromolecular and cellular level at endosseous implant/biosystem interfaces. However, we are still far away from designing sophisticated implant surfaces with the best possible, selective functionality for each specific tissue or cavity interface. Firstly, because our knowledge of the respective surface related reactions is at best fragmentary. Secondly, because manufacturing of multi-scaled complex surfaces including distinct nanotopographies, wetting properties, and stable cleanliness is still a technical challenge and far away from being reproducibly transferred to implant surfaces.

Coating of ß-tricalcium phosphate scaffolds-a comparison between graphene oxide and poly-lactic-co-glycolic acid.

Bone regeneration in critical size defects is a major challenge in oral and maxillofacial surgery, and the gold standard for bone reconstruction still requires the use of autologous tissue. To overcome the need for a second intervention and to minimize morbidity, the development of new biomaterials with osteoinductive features is the focus of current research. As a scaffolding material, ß-tricalcium phosphate (ß-TCP) is suitable for bone regeneration purposes, although it does not carry any functional groups for the covalent immobilization of molecules. The aim of the present study was to establish effective coating variants for ß-TCP constructs to enable the biofunctionalization of anorganic blocks with different osteogenic molecules in future studies. We established working protocols for thin surface coatings consisting of polylactic-co-glycolic acid (PLGA) and graphene oxide (GO) by varying parameters. Surface properties such as the angularity and topography of the developed scaffolds were analyzed. To examine biological functionality, the adhesion and proliferation behavior of jaw periosteal cells (JPCs) were tested on the coated constructs. Our results suggest that PLGA is the superior material for surface coating of ß-TCP matrices, leading to higher JPC proliferation rates and providing a more suitable basis for further biofunctionalization in the field of bone tissue engineering.

Comparison of different in vitro tests for biocompatibility screening of Mg alloys.

Standard cell culture tests according to ISO 10993 have only limited value for the biocompatibility screening of degradable biomaterials such as Mg alloys. The correlation between in vitro and in vivo results is poor. Standard cytotoxicity tests mimic the clinical situation to only a limited extent, since in vivo proteins and macromolecules in the blood and interstitial liquid will influence the corrosion behaviour and, hence, biocompatibility of Mg alloys to a significant extent. We therefore developed a modified cytotoxicity test simulating the in vivo conditions by use of bovine serum as the extraction vehicle instead of the cell culture medium routinely used in standard cytotoxicity testing according to ISO 10993-5. The modified extraction test was applied to eight experimental Mg alloys. Cytotoxicity was assayed by inhibition of cell metabolic activity (XTT test). When extraction of the alloy samples was performed in serum instead of cell culture medium the metabolic activity was significantly less inhibited for six of the eight alloys. The reduction in apparent cytotoxicity under serum extraction conditions was most pronounced for MgZn1 (109% relative metabolic activity with serum extracts vs. 26% in Dulbecco's modified Eagle's medium (DMEM)), for MgY4 (103% in serum vs. 32% in DMEM) and for MgAl3Zn1 (84% vs. 17%), resulting in a completely different cytotoxicity ranking of the tested materials when serum extraction was used. We suppose that this test system has the potential to enhance the predictability of in vivo corrosion behaviour and biocompatibility of Mg-based materials for biodegradable medical devices.

Formation and photocatalytic decomposition of a pellicle on anatase surfaces.

The acquired dental pellicle plays a critical role in the adhesion and detachment of dental plaque bacteria. It has been reported that titanium dioxide biomaterials decompose single-protein films by photocatalysis. However, it is not known whether this can also be achieved with complex structured pellicle films. This in vitro study investigated in real-time the formation and photocatalytic decomposition of human pellicle at anatase-saliva interfaces. Nanostructured polycrystalline anatase layers were deposited on titanium-coated quartz crystals by magnetron-sputtering, serving as a model for titanium implant surfaces. The quartz crystals were used as acoustic sensors in a quartz-crystal microbalance (QCM) system with dissipation. In situ UV irradiation of pellicle-covered anatase caused a statistically significant decrease of the adsorbed salivary mass. In contrast, photocatalytic decomposition of pellicle could not be observed on reference titanium surfaces. Wettability characterization revealed superhydrophilicity of anatase upon UV irradiation, whereas titanium was unaffected. XPS measurements provide further information concerning the decomposition of the salivary films. The results suggest that the photocatalytic activity of polycrystalline anatase-modified biomaterial surfaces is able to decompose complex structured macromolecular pellicle films. Therefore, this study opens the way to surface modifications supporting therapeutic approaches of biofilm removal.

Multifunctional nature of UV-irradiated nanocrystalline anatase thin films for biomedical applications.

Anatase is known to decompose organic material by photocatalysis and to enhance surface wettability once irradiated by ultraviolet (UV) light. In this study, pulse magnetron-sputtered anatase thin films were investigated for their suitability with respect to specific biomedical applications, namely superhydrophilic and biofilm degrading implant surfaces. UV-induced hydrophilicity was quantified by static and dynamic contact angle analysis. Photocatalytic protein decomposition was analyzed by quartz crystal microbalance with dissipation. The surfaces were characterized by X-ray diffraction, atomic force microscopy, scanning electron microscopy and X-ray photoelectron spectroscopy. The radical formation on anatase, responsible for photocatalytic effects, was analyzed by electron spin resonance spectroscopy. Results have shown that the nanocrystalline anatase films, in contrast to reference titanium surfaces, were sensitive to UV irradiation and showed rapid switching towards superhydrophilicity. The observed decrease in carbon adsorbents and the increase in the fraction of surface hydroxyl groups upon UV irradiation might contribute to this hydrophilic behavior. UV irradiation of anatase pre-conditioned with albumin protein layers induces the photocatalytic decomposition of these model biofilms. The observed degradation is mainly caused by hydroxyl radicals. It is concluded that nanocrystalline anatase films offer different functions at implant interfaces, e.g. bedside hydrophilization of anatase-coated implants for improved osseointegration or the in situ decomposition of conditioning films forming the basal layer of biofilms in the oral cavity.

[Effects of voltage during anodization on titanium surface on cell attachment and spreading of osteoblasts].

PURPOSE: The effects of the voltage during anodic oxidation on cell attachment and spreading of human osteoblasts in early stage were investigated. METHODS: The samples were anodized in the electrolyte of 0.03 M calcium glycerophosphate (Ca-GP) mixed with 0.15 M calcium acetate (CA) under current density of 70 A/m2 and different voltages, 140V, 200V or 260V. The surface roughness of the specimens and the attachment and spreading of human osteoblasts in early stage were investigated. SPSS13.0 for windows was used for one-way ANOVA. RESULTS: The surface average roughness of cp Ti was significantly improved from 0.17 micro m to 0.23 micro m, 0.26 micro m, 0.33 micro m, respectively, by anodic oxidation with increased voltage (P<0.05). After 2 hours of cell culture, cell skeleton was reorganized and cell morphology became more irregular with the increase of anodizing voltages. Cell attachment on the specimens anodized under 260V was significantly higher than on the surface of cp Ti (P<0.05). CONCLUSION: The surface properties of cp Ti can be affected by anodic oxidation. The early cell attachment and spreading were enhanced with anodizing voltage.

Enhancing surface free energy and hydrophilicity through chemical modification of microstructured titanium implant surfaces.

Roughness-induced hydrophobicity, well-known from natural plant surfaces and intensively studied toward superhydrophobic surfaces, has currently been identified on microstructured titanium implant surfaces. Studies indicate that microstructuring by sandblasting and acid etching (SLA) enhances the osteogenic properties of titanium. The undesired initial hydrophobicity, however, presumably decelerates primary interactions with the aqueous biosystem. To improve the initial wettability and to retain SLA microstructure, a novel surface modification was tested. This modification differs from SLA by its preparation after acid etching, which was done under protective gas conditions following liquid instead of dry storage. We hypothesized that this modification should have increased wettability due to the prevention of contaminations that occurs during air contact. The main outcome of dynamic wettability measurements was that the novel modification shows increased surface free energy (SFE) and increased hydrophilicity with initial water contact angles of 0 degrees compared to 139.9 degrees for SLA. This hydrophilization was kept even after any drying. Reduced hydrocarbon contaminations were identified to play a possible role in altered surface thermodynamics. Such surfaces aim to retain the hydrophilicity and natural high surface energy of the Ti dioxide surface until surgical implants' insertion and are compared in this in vitro study with structural surface variants of titanium to compare roughness and chemically induced wettability.

Roughness induced dynamic changes of wettability of acid etched titanium implant modifications.

Dynamic contact angle analysis (DCA) was used to investigate time-dependent wettability changes of sandblasted and acid-etched commercially pure (cp) titanium (Ti) implant modifications during their initial contact with aqueous systems compared to a macrostructured reference surface. Surface topography was analyzed by scanning electron microscopy and by contact stylus profilometry. The microstructured Ti surfaces were found to be initially extremely hydrophobic. This hydrophobic configuration can shift to a completely wettable surface behavior with water contact angles of 0 degrees after the first emersion loop during DCA experiments. It is suggested that a hierarchically structured surface topography could be responsible for this unexpected wetting phenomenon. Roughness spatial and hybrid parameters could describe topographical features interfering with dynamic wettability significantly better than roughness height parameters. The Ti modifications which shift very sudden from a hydrophobic to a hydrophilic state adsorbed the highest amount of immunologically assayed fibronectin. The results suggest that microstructuring greatly influences both the dynamic wettability of Ti implant surfaces during the initial host contact and the initial biological response of plasma protein adsorption. The microstructured surfaces, once in the totally wettable configuration, may improve the initial contact with host tissue after implantation, due to the drastically increased hydrophilicity.

Detection of nonvolatile macromolecules in breath. A possible diagnostic tool?

The analysis of parameters in bronchoalveolar extracellular lining secretions has come into greater use in the diagnosis of diseases of the lung and respiratory passages. The bronchoalveolar lavage (BAL) method is thus used for sampling alveolar fluids or bronchial secretions. However, this method is invasive and therefore cannot be routinely employed for probe sampling. Based on the hypothesis that aerosol particles excreted in human breath reflect the composition of the bronchoalveolar extracellular lining fluid, experiments were performed to concentrate and analyze these aerosols directly using a noninvasive technique. Human exhaled air was directed through a set of cool traps and the condensate of 200 to 400 exhalations examined for nonvolatile components, such as proteins. In experiments conducted with volunteers, the amount of proteins in the breath condensate of 8 healthy individuals (of a total of 10) amounted to between 4 micrograms and 1.4 mg. The proteins were separated by two-dimensional polyacrylamide gel electrophoresis (PAGE) and compared to saliva samples of the respective volunteers. The results suggest that the proteins detected in breath originate partially from the naso-oropharyngeal tract and partially from lower regions of the airways. In clinical tests, the exhaled air of 13 patients suffering from various diseases of the respiratory tract was sampled and analyzed by immunoassays for inflammation parameters, such as interleukin-1 beta (IL-1 beta), soluble interleukin-2 receptor protein, light chain (sIL-2R), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-alpha). In these tests, up to 370 pg IL-1 beta, 120 pg TNF-alpha, and 2,159 U sIL-2R per ml were measured in the breath condensate.(ABSTRACT TRUNCATED AT 250 WORDS)

Method for identification of intracellular free flavin species in the photosensitive fungus Neurospora crassa.

Establishing the relative intracellular proportions of flavins in Neurospora crassa (and in other organisms) in vivo may be hampered by degradation of flavins after homogenization of the cells. The system described here allows separation and identification of intracellular free and bound flavins under conditions restrictive for the FAD-degrading enzyme(s). A "protective buffer" containing 0.1 M citrate adjusted to pH 4.0 with K2HPO4, 5 mM ATP, and 0.5 mM EDTA prevents FAD from rapid enzymatic cleavage in crude cell lysates of the Neurospora crassa mutant "slime."

Nitrate reductase activity test: phenazine methosulfate-ferricyanide stop reagent replaces postassay treatment.

Nitrate reductase activity is usually measured by colorimetric determination of the nitrite formed. Since reduced pyridine nucleotides interfere with color formation, the use of NADPH or NADH in the assay requires a specific postassay treatment to remove excess substrate. A "stop mix" containing 1.5 mM phenazine methosulfate and 4.0 mM ferricyanide (final concentrations 0.136 and 0.36 mM, respectively) can remove excess NAD(P)H and terminate the enzymatic reaction quickly in a single, time-saving step. For activity tests containing dithionite we recommend the use of a 1:1 mixture of the two color reagents to avoid incomplete color formation. This may occur during longer time intervals between addition of the color reagents due to destruction of the diazonium salt formed with the first reagent by oxidation product(s) of dithionite.