Collagen-Based Osteogenic Nanocoating of Microrough Titanium Surfaces.

The aim of the present study was to develop a collagen/heparin-based multilayer coating on titanium surfaces for retarded release of recombinant human bone morphogenic protein 2 (rhBMP2) to enhance the osteogenic activity of implant surfaces. Polyelectrolyte multilayer (PEM) coatings were constructed on sandblasted/acid-etched surfaces of titanium discs using heparin and collagen. PEM films of ten double layers were produced and overlayed with 200 µL of a rhBMP2 solution containing 15 µg rhBMP2. Subsequently, cross-linking of heparin molecules was performed using EDC/NHS chemistry to immobilize the incorporated rhBMP2. Release characteristics for 3 weeks, induction of Alkaline Phosphatase (ALP) in C2C12 cells and proliferation of human mesenchymal stem cells (hMSCs) were evaluated to analyze the osteogenic capacity of the surface. The coating incorporated 10.5 µg rhBMP2 on average per disc and did not change the surface morphology. The release profile showed a delivery of 14.5% of the incorporated growth factor during the first 24 h with a decline towards the end of the observation period with a total release of 31.3%. Cross-linking reduced the release with an almost complete suppression at 100% cross-linking. Alkaline Phosphatase was significantly increased on day 1 and day 21, indicating that the growth factor bound in the coating remains active and available after 3 weeks. Proliferation of hMSCs was significantly enhanced by the non-cross-linked PEM coating. Nanocoating using collagen/heparin-based PEMs can incorporate clinically relevant amounts of rhBMP2 on titanium surfaces with a retarded release and a sustained enhancement of osteogenic activity without changing the surface morphology.

Poly-Alanine-ε-Caprolacton-Methacrylate as Scaffold Material with Tuneable Biomechanical Properties for Osteochondral Implants.

An aging population and injury-related damage of the bone substance lead to an increasing need of innovative materials for the regeneration of osteochondral defects. Biodegradable polymers form the basis for suitable artificial implants intended for bone replacement or bone augmentation. The great advantage of these structures is the site-specific implant design, which leads to a considerable improvement in patient outcomes and significantly reduced post-operative regeneration times. Thus, biomechanical and biochemical parameters as well as the rate of degradation can be set by the selection of the polymer system and the processing technology. Within this study, we developed a polymer platform based on the amino acid Alanine and ε-Caprolacton for use as raw material for osteochondral implants. The biomechanical and degradation properties of these Poly-(Alanine-co-ε-Caprolacton)-Methacrylate (ACM) copolymers can be adjusted by changing the ratio of the monomers. Fabrication of artificial structures for musculo-skeletal tissue engineering was done by Two-Photon-Polymerization (2PP), which represents an innovative technique for generating defined scaffolds with tailor-made mechanical and structural properties. Here we show the synthesis, physicochemical characterization, as well as first results for structuring ACM using 2PP technology. The data demonstrate the high potential of ACM copolymers as precursors for the fabrication of biomimetic implants for bone-cartilage reconstruction.

Porous 3D Scaffolds Enhance MSC Vitality and Reduce Osteoclast Activity.

In the context of an aging population, unhealthy Western lifestyle, and the lack of an optimal surgical treatment, deep osteochondral defects pose a great challenge for the public health system. Biodegradable, biomimetic scaffolds seem to be a promising solution. In this study we investigated the biocompatibility of porous poly-((D,L)-lactide-ε-caprolactone)dimethacrylate (LCM) scaffolds in contrast to compact LCM scaffolds and blank cell culture plastic. Thus, morphology, cytotoxicity and metabolic activity of human mesenchymal stromal cells (MSC) seeded directly on the materials were analyzed after three and six days of culturing. Further, osteoclastogenesis and osteoclastic activity were assessed using reverse-transcriptase real-time PCR of osteoclast-specific genes, EIA and morphologic aspects after four, eight, and twelve days. LCM scaffolds did not display cytotoxic effects on MSC. After three days, metabolic activity of MSC was enhanced on 3D porous scaffolds (PS) compared to 2D compact scaffolds (CS). Osteoclast activity seemed to be reduced at PS compared to cell culture plastic at all time points, while no differences in osteoclastogenesis were detectable between the materials. These results indicate a good cytocompatibility of LCM scaffolds. Interestingly, porous 3D structure induced higher metabolic activity of MSC as well as reduced osteoclast activity.

Two-Photon Polymerized Poly(2-Ethyl-2-Oxazoline) Hydrogel 3D Microstructures with Tunable Mechanical Properties for Tissue Engineering.

The main task of tissue engineering (TE) is to reproduce, replicate, and mimic all kinds of tissues in the human body. Nowadays, it has been proven useful in TE to mimic the natural extracellular matrix (ECM) by an artificial ECM (scaffold) based on synthetic or natural biomaterials to regenerate the physiological tissue/organ architecture and function. Hydrogels have gained interest in the TE community because of their ability to absorb water similar to physiological tissues, thus mechanically simulating the ECM. In this work, we present a novel hydrogel platform based on poly(2-ethyl-2-oxazoline)s, which can be processed to 3D microstructures via two-photon polymerization (2PP) with tunable mechanical properties using monomers and crosslinker with different degrees of polymerization (DP) for future applications in TE. The ideal parameters (laser power and writing speed) for optimal polymerization via 2PP were obtained using a specially developed evaluation method in which the obtained structures were binarized and compared to the computer-aided design (CAD) model. This evaluation was performed for each composition. We found that it was possible to tune the mechanical properties not only by application of different laser parameters but also by mixing poly(2-ethyl-2-oxazoline)s with different chain lengths and variation of the crosslink density. In addition, the swelling behavior of different fabricated hydrogels were investigated. To gain more insight into the viscoelastic behavior of different fabricated materials, stress relaxation tests via nanoindentation experiments were performed. These new hydrogels can be processed to 3D microstructures with high structural integrity using optimal laser parameter settings, opening a wide range of application properties in TE for this material platform.

Aging of plasma-polymerized allylamine nanofilms and the maintenance of their cell adhesion capacity.

The long-term stability and γ-sterilisability of bioactive layers is the precondition for the application of implants. Thus, aging processes of a microwave deposited, plasma polymerized allylamine nanofilm (PPAAm) with positively charged amino groups were evaluated concerning physicochemical characteristics and cell adhesion capacity over the course of one year. XPS, FT-IR, surface free energy, and water contact angle measurements elucidated not only the oxidation of the PPAAm film due to atmospheric oxygen reacting with surface free radicals but also the influence of atmospheric moisture during sample storage in ambient air. Surprisingly, within 7 days 70% of the primary amino groups are lost and mostly converted into amides. A positive zeta-potential was verified for half a year and longer. Increasing polar surface groups and a water contact angle shift from 60° to 40° are further indications of altered surface properties. Nevertheless, MG-63 human osteoblastic cells adhered and spread out considerably on aged and additionally γ-sterilized PPAAm layers deposited on polished titanium alloys (Ti-6Al-4V_P). These cell-relevant characteristics were highly significant over the whole period of one year and may not be related to the existence of primary amino groups. Rather, the oxidation products, the chemical amide group, that is, seem to support the attachment of osteoblasts at all times up to one year.

Sclerostin Alters Tumor Cell Characteristics of Oral Squamous Cell Carcinoma and May Be a Key Player in Local Bone Invasion.

Localized jawbone invasion is a milestone in the progression of oral squamous cell carcinoma (OSCC). The factors that promote this process are not well understood. Sclerostin is known to be involved in bone metabolism and there are preliminary reports of its involvement in bone tumors and bone metastasis. To identify a possible involvement of sclerostin in the bone invasion process of OSCC, sclerostin expression was analyzed in vitro in two different human OSCC tumor cell lines by quantitative real-time polymerase chain reaction (qRT-PCR), and the effect of recombinant human (rh)-sclerostin treatment on tumor cell capabilities was evaluated using proliferation, migration, and invasion assays. Undifferentiated human mesenchymal stem cells (hMSCs) were osteogenically differentiated and co-cultured with OSCC tumor cells to demonstrate potential interactions and migration characteristics. Sclerostin expression was evaluated in clinical cases by immunohistochemistry at the OSCC-jawbone interface in a cohort of 15 patients. Sclerostin expression was detected in both OSCC tumor cell lines in vitro and was also detected at the OSCC-jawbone interface in clinical cases. Tumor cell proliferation rate, migration and invasion ability were increased by rh-sclerostin treatment. The migration rate of tumor cells co-cultured with osteogenically differentiated hMSCs was increased. The results presented are the first data suggesting a possible involvement of sclerostin in the bone invasion process of OSCC, which deserves further investigation and may be a potential approach for drug-based tumor therapy.

Dependence of the initial adhesion of biofilm forming Pseudomonas putida mt2 on physico-chemical material properties.

Bacterial adhesion is strongly dependent on the physico-chemical properties of materials and plays a fundamental role in the development of a growing biofilm. Selected materials were characterized with respect to their physico-chemical surface properties. The different materials, glass and several polymer foils, showed a stepwise range of surface tensions (γ(s)) between 10.3 and 44.7 mN m(-1). Measured zeta potential values were in the range between -74.8 and -28.3 mV. The initial bacterial adhesion parameter q(max) was found to vary between 6.6 × 10(6) and 28.1 × 10(6) cm(-2). By correlation of the initial adhesions kinetic parameters with the surface tension data, the optimal conditions for the immobilization of Pseudomonas putida mt2 were found to be at a surface tension of 24.7 mN m(-1). Both higher and lower surface tensions lead to a smaller number of adherent cells per unit surface area. Higher energy surfaces, commonly termed hydrophilic, could constrain bacterial adhesion because of their more highly ordered water structure (exclusion zone) close to the surface. At low energy surfaces, commonly referred to as hydrophobic, cell adhesion is inhibited due to a thin, less dense zone (depletion layer or clathrate structure) close to the surface. Correlation of q (max) with zeta potential results in a linear relationship. Since P. putida carries weak negative charges, a measurable repulsive effect can be assumed on negative surfaces.

Development of a system of heparin multilayers on titanium surfaces for dual growth factor release.

The aim of the present study was to establish a modular platform of poly-L-lysine-heparin (PLL-Hep) polyelectrolyte multilayer (PEM) coatings on titanium surfaces for dual growth factor delivery of recombinant human bone morphogenic protein 2 (rhBMP2) and recombinant human vascular endothelial growth factor 165 (rhVEGF165) in clinically relevant quantities. Release characteristics for both growth factors differed significantly depending on film architecture. rhBMP2 induced activation of alkaline phosphatase in C2C12 cells and proliferation of human mesenchymal stem cells (hMSCs). rhVEGF mediated induction of von Willebrand factor (vWF) in hMSCs and proliferation of human umbilical vein endothelial cells. Osteogenic and angiogenic effects were modified by variation in cross-linking and architecture of the PEMs. By creating multilayer films with distinct zones, release characteristics and proportion of both growth factor delivery could be tuned and surface-activity modified to enhance angiogenic or osteogenic function in various ways. In summary, the system provides a modular platform for growth factor delivery that allows for individual composition and accentuation of angiogenic and osteogenic surface properties.

Tailored Polyelectrolyte Multilayer Systems by Variation of Polyelectrolyte Composition and EDC/NHS Cross-Linking: Physicochemical Characterization and In Vitro Evaluation.

The layer-by-layer (LbL) self-assembly technique is an effective method to immobilize components of the extracellular matrix (ECM) such as collagen and heparin onto, e.g., implant surfaces/medical devices with the aim of forming polyelectrolyte multilayers (PEMs). Increasing evidence even suggests that cross-linking influences the physicochemical character of PEM films since mechanical cues inherent to the substrate may be as important as its chemical nature to influence the cellular behavior. In this study, for the first-time different collagen/heparin films have been prepared and cross-linked with EDC/NHS chemistry. Quartz crystal microbalance, zeta potential analyzer, diffuse reflectance Fourier transform infrared spectroscopy, atomic force microscopy and ellipsometry were used to characterize film growth, stiffness, and topography of different film systems. The analysis of all data proves a nearly linear film growth for all PEM systems, the efficacy of cross-linking and the corresponding changes in the film rigidity after cross-linking and an appropriate surface topography. Furthermore, preliminary cell culture experiments illustrated those cellular processes correlate roughly with the quantity of newly created covalent amide bonds. This allows a precise adjustment of the physicochemical properties of the selected film architecture regarding the desired application and target cells. It could be shown that collagen improves the biocompatibility of heparin containing PEMs and due to their ECM-analogue nature both molecules are ideal candidates intended to be used for any biomedical application with a certain preference to improve the performance of bone implants or bone augmentation strategies.

Tailored Polyelectrolyte Multilayer Systems by Variation of Polyelectrolyte Composition and EDC/NHS Cross-Linking: Controlled Drug Release vs. Drug Reservoir Capabilities and Cellular Response for Improved Osseointegration.

Polyelectrolyte multilayers (PEM) are versatile tools used to investigate fundamental interactions between material-related parameters and the resulting performance in stem cell differentiation, respectively, in bone tissue engineering. In the present study, we investigate the suitability of PEMs with a varying collagen content for use as drug carriers for the human bone morphogenetic protein 2 (rhBMP-2). We use three different PEM systems consisting either of the positively charged poly-L-lysine or the glycoprotein collagen type I and the negatively charged glycosaminoglycan heparin. For a specific modification of the loading capacity and the release kinetics, the PEMs were stepwise cross-linked before loading with cytokine. We demonstrate the possibility of immobilizing significant amounts of rhBMP-2 in all multilayer systems and to specifically tune its release via cross-linking. Furthermore, we prove that the drug release of rhBMP-2 plays only a minor role in the differentiation of osteoprogenitor cells. We find a significantly higher influence of the immobilized rhBMP-2 within the collagen-rich coatings that obviously represent an excellent mimicry of the native extracellular matrix. The cytokine immobilized in its bioactive form was able to achieve an increase in orders of magnitude both in the early stages of differentiation and in late calcification compared to the unloaded layers.

Bacterial nanocellulose: Reinforcement of compressive strength using an adapted Mobile Matrix Reservoir Technology and suitable post-modification strategies.

Bacterial nanocellulose (BNC) is a highly interesting biomaterial due to some outstanding properties especially when used in medical therapeutics and diagnostics. BNC is absolutely bioinert and is characterised by intrinsic properties such as high tensile stiffness and elasticity, high porosity, exceptional water uptake and swelling capacity. Furthermore, these properties can be adjusted in a very defined way by specifically changing the cultivation conditions or performing post-modifications such as crosslinking, functionalisation with additives, dehydration or drying. Especially the high tensile strength of the nanofibrillar material has been the subject of many investigations in the past couple of years. Nevertheless, the enormous tensile strength and elasticity of BNC is contrary to an almost purely viscous behaviour under compressive load. In the present study, different methods to influence the mechanical behaviour under compression with respect to load bearing applications of BNC are systematically investigated. The possibilities and limitations of the variable layer-by-layer cultivation known as Mobile Matrix Reservoir Technology (MMR-Tech) as well as the effect of different post-modification strategies of BNC are thoroughly investigated. Beside of commonly used indentation tests for characterising the mechanical properties of BNC, we introduce a novel evaluation methodology based on mechanical relaxation measurements and an evolutionary regression algorithm for the derivation of a viscoelastic material law, which for the first time allows standardised, comparative viscoelastic investigations of soft-matter biomaterials to be performed independently of the measurement setup. Using this methodology, we are able to show, that cultivation conditions for BNC and suitable post-modifications can result in different effects on the viscoelastic behaviour of the fabricated composites. We show that the cultivation conditions for BNC primarily affect the height of dispersion and the frequency of the relaxation centre which corresponds roughly to the mean value of the logarithmic distributed relaxation times, and that these effects could be enhanced by post-modifications. However, we also identify parameters, such as the width of the relaxation region, which corresponds roughly to the standard deviation of the logarithmic distributed relaxation times, on which the type of cultivation obviously shows no influence but which can be influenced exclusively by post-modifications. Our methodology enables for the first time a clear identification of those parameters which represent a significant factor of influence to the viscoelastic material behaviour, which should enable a more targeted and application-relevant development of BNC composites in the future.

Colloidal force spectroscopy and cell biological investigations on biomimetic polyelectrolyte multilayer coatings composed of chondroitin sulfate and heparin.

To promote osteoblast adhesion and proliferation on (bio)material surfaces, biomimetic coatings resembling the natural extracellular matrix (ECM) are desirable. The glycosamino glycans (GAGs) chondroitin sulfate (CS) and heparin (HEP) are promising candidates for a biomimetic coating since they are two of the most prevalent noncollagenous biomolecules constituting the ECM. Coatings containing CS and HEP were prepared employing the "layer by layer" technique yielding polyelectrolyte multilayers (PEMs). Physicochemical and mechanical characterization of the coatings were performed by means of streaming potential measurements and colloidal force spectroscopy. The capability of the coatings to support cell adhesion, spreading, proliferation, and maintenance of an osteoblastic phenotype was assessed with SaOS osteosarcoma cells. We demonstrate that PEMs constructed from CS as the polyanion display a low Young's modulus correlated with poorly supported cell adhesion and proliferation. When the CS was adsorbed onto a stiffer polypeptide PEM basis, the Young's modulus increased, and the cell response was significantly improved. For HEP coatings an intermediate Young's modulus and moderate cell adhesion and spreading were observed. No significant changes in stiffness or cell response were detected when HEP was adsorbed onto the polypeptide film.

First Isolation and Structure Elucidation of GDNT-ß-Glu - Tetraether Lipid Fragment from Archaeal Sulfolobus Strains.

Due to their special chemical structure, tetraether lipids (TEL) represent essential elements of archaeal membranes, providing these organisms with extraordinary properties. Here we describe the characterization of a newly isolated structural element of the main lipids. The TEL fragment GDNT-ß-Glu was isolated from Sulfolobus metallicus and characterized in terms of its chemical structure by NMR- and MS-investigations. The obtained data are dissimilar to analogically derived established structures - in essence, the binding relationships in the polar head group are re-determined and verified. With this work, we provide an important contribution to the structure elucidation of intact TEL also contained in other Sulfolobus strains such as Solfulobus acidocaldarius and Sulfolobus solfataricus.

Two-photon techniques in tissue engineering.

PURPOSE: NIR radiation in the range of about 800 nm is less absorbed by biological tissues and is suited for triggering photonic effects using femtosecond pulsed Ti:Sa lasers. Especially in the life sciences, two-photon techniques are gaining greater importance. We introduce two laser applications for tissue engineering: the autofluorescent visualization of cells seeded on 3D scaffolds after two-photon excitation; and the manufacturing of 3D-structured hydrogel-like scaffolds by triggering free-radical polymerization processes within polymerizable precursors. METHODS: Primary bovine chondrocytes were cultivated on collagen I/III scaffolds using a flow chamber system coupled with a two-photon laser scanning microscope (2PLSM). During the incubation the cell population was hydrostatically stimulated. The selective visualization of unlabeled cells and scaffolds was achieved by spectral autofluorescence imaging. To gain some insight into scaffold-mediated effects on cell growth and cell differentiation, hydrogel-like scaffolds with well defined 3D structures were generated by two-photon polymerization (2PP) using methacrylated urethane and polyethyleneglycol diacrylate. RESULTS: We were able to show that spectral autofluorescence imaging provides spatially resolved data for the non-invasive online control of the tissue engineering process as well as the quantification of cell distribution within the scaffold. The fabrication of 3D 2PP scaffolds made from hydrogel-forming monomers and their effect on cell attachment and cell growth were also shown. CONCLUSIONS: Two-photon techniques provide powerful tools for both the non-invasive online visualization of 3D cell-scaffold constructs and the structuring of 3D cultivation environments. The application of these techniques is also suitable for integration into micro-systems technology (e.g. BioMEMS, Cells-on-Chip, Lab-on-a Chip).

Modulation of Platelet-Surface Activation: Current State and Future Perspectives.

Modulation of platelet-surface activation is important for many biomedical applications such as in vivo performance, platelet storage, and acceptance of an implant. Reducing platelet-surface activation is challenging because they become activated immediately after short contact with nonphysiological surfaces. To date, controversies and open questions in the field of platelet-surface activation still remain. Here, we review state-of-the-art approaches in inhibiting platelet-surface activation, mainly focusing on modification, patterning, and methodologies for characterization of the surfaces. As a future perspective, we discuss how the combination of biochemical and physiochemical strategies together with the topographical modulations would assist in the search for an ideal nonthrombogenic surface.

Biomimetic multilayer coatings deliver gentamicin and reduce implant-related osteomyelitis in rats.

Implant-related infections like periprosthetic joint infections (PJI) are still a challenging issue in orthopedic surgery. In this study, we present a prophylactic anti-infective approach based on a local delivery of the antibiotic gentamicin. The local delivery is achieved via a nanoscale polyelectrolyte multilayer (PEM) coating that leaves the bulk material properties of the implant unaffected while tuning the surface properties. The main components of the coating, i.e. polypeptides and sulfated glycosaminoglycans (sGAG) render this coating both biomimetic (matrix mimetic) and biodegradable. We show how adaptions in the conditions of the multilayer assembly process and the antibiotic loading process affect the amount of delivered gentamicin. The highest concentration of gentamicin could be loaded into films composed of polypeptide poly-glutamic acid when the pH of the loading solution was acidic. The concentration of gentamicin on the surface could be tailored with the number of deposited PEM layers. The resulting coatings reveal a bacteriotoxic effect on Staphylococcus cells but show no signs of cytotoxic effects on MC3T3-E1 osteoblasts. Moreover, when multilayer-coated titanium rods were implanted into contaminated medullae of rat tibiae, a reduction in the development of implant-related osteomyelitis was observed. This reduction was more pronounced for the multifunctional, matrix-mimetic heparin-based coatings that only deliver lower amounts of gentamicin.

Biomimetic Designer Scaffolds Made of D,L-Lactide-ɛ-Caprolactone Polymers by 2-Photon Polymerization.

IMPACT STATEMENT: In tissue engineering (TE), the establishment of cell targeting materials, which mimic the conditions of the physiological extracellular matrix (ECM), seems to be a mission impossible without advanced materials and fabrication techniques. With this in mind we established a toolbox based on (D,L)-lactide-ɛ-caprolactone methacrylate (LCM) copolymers in combination with a nano-micromaskless lithography technique, the two-photon polymerization (2-PP) to mimic the hierarchical structured and complex milieu of the natural ECM. To demonstrate the versatility of this toolbox, we choose two completely different application scenarios in bone and tumor TE to show the high potential of this concept in therapeutic and diagnostic application.

Archaeal tetraether lipid coatings-A strategy for the development of membrane analog spacer systems for the site-specific functionalization of medical surfaces.

The primary goal of our investigation was the development of a versatile immobilization matrix based on archaeal tetraether lipids that meets the most important prerequisites to render an implant surface bioactive by binding specific functional groups or functional polymers with the necessary flexibility and an optimal spatial arrangement to be bioavailable. From this point of view, it appears obvious that numerous efforts made recently to avoid initial bacterial adhesion on catheter surfaces as an important prerequisite of material associated infection episodes have shown only a limited efficiency since the bioactive entities could not be presented in an optimal conformation and a stable density. A significant improvement of this situation can be achieved by highly specific biomimetic modifications of the catheter surfaces. The term "biomimetic" originates from the fact that specific archaeal tetraether lipids were introduced to form a membrane analog monomolecular spacer system, which (1) can be immobilized on nearly all solid surfaces and (2) chemically modified to present a tailor-made functionality in contact with aqueous media either to avoid or inhibit surface fouling or to equip any implant surface with the necessary chemical functionality to enable cell adhesion and tissue integration. Ultrathin films based on tetraether lipids isolated from archaea Thermoplasma acidophilum were used as a special biomimetic immobilization matrix on the surface of commercial medical silicon elastomers. A complete performance control of the membrane analog coatings was realized in addition to biofunctionality tests, including the proof of cytotoxicity and hemocompatibility according to DIN EN ISO 10993. In order to make sure that the developed immobilization matrix including the grafted functional groups are biocompatible under in vivo-conditions, specific animal tests were carried out to examine the in vivo-performance. It can be concluded that the tetraether lipid based coating systems on silicone have shown no signs of cytotoxicity and a good hemocompatibility. Moreover, no mutagenic effects, no irritation effects, and no sensitization effects could be demonstrated. After an implantation period of 28 days, no irregularities were found.

Improved initial osteoblast functions on amino-functionalized titanium surfaces.

Adhesion and spreading of cells on biomaterials are integrin-mediated processes. But recent findings indicate a key role of the cell membrane associated matrix substance hyaluronan (HA) in interface interactions. Because HA is a negatively charged molecule we assume that a biomaterial surface with an opposed charge could boost the first contact of the cell to the surface. Polished cp titanium (R(a)=0.19 microm) was coated with an amino-group containing plasma polymer (Ti PPA). For this purpose, a microwave excited, pulsed, low-pressure plasma was used. Additionally, collagen was immobilized on Ti PPA with polyethylene glycol diacid (PEG-DA), catalyzed by carbodiimide (CDI). The physico-chemical surface analytical techniques like XPS, FT-IR, water contact angle and zeta-potential verified the retention of the allylamine precursor structure. Human osteoblasts were cultured in serum-free Dulbecco's modified Eagle medium (DMEM). Adhesion and cell cycle phases were calculated by flow cytometry. Spreading and actin cytoskeleton were visualized by confocal microscopy. Gene expression of osteogenic markers was detected by real-time RT-PCR. Ti PPA is significantly advantageous concerning initial adhesion and spreading during the first hours of the cell contact to the surface. The proliferation of osteoblasts is positively influenced. Gene expression of the differentiation marker bone sialoprotein was upregulated after 24h. Our results demonstrate that functionalization of titanium with positively charged amino-groups is sufficiently enough to significantly improve initial steps of the cellular contact to the material surface.

Antibacterial and anti-encrustation biodegradable polymer coating for urinary catheter.

Bacterial biofilm and crystalline deposits are the common causes of failure of long-term indwelling urinary catheter. Bacteria colonise the catheter surface causing serious infections in the urinary tract and encrustations that can block the catheter and induce trauma in patients. In this study, the strategy used to resist bacterial adhesion and encrustation represents a combination of the antibacterial effects of norfloxacin and silver nanoparticles and the PLGA-based neutralisation of alkali products of urea hydrolysis gained through the degradation of the polymer in an aqueous milieu. Silver nanoparticles were coated with tetraether lipids (TEL) to avoid aggregation when dispersed in acetone and during the film formation. The polymer films loaded with the two antibacterial agents were applied on Polyurethane (PUR) and Silicon sheets. We demonstrated the antibacterial and anti-adhesion effectiveness of the coatings whereby commercially available biocompatible polymers PUR and Silicon were used as controls. Using artificial urine and an in vitro encrustation model, it was shown that the coatings resist the encrustation for at least 2 weeks. This combination of a biodegradable polymer and wide-range antibacterial agents represents a potentially attractive biocompatible coating for urinary catheters.