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Surface biofunctionalization by covalent co-immobilization of oligopeptides.

Functionalization of implants with multiple bioactivities is desired to obtain surfaces with improved biological and clinical performance. Our objective was developing a simple and reliable method to obtain stable multifunctional coatings incorporating different oligopeptides. We co-immobilized on titanium surface oligopeptides of known cooperative bioactivities with a simple and reliable method. Appropriately designed oligopeptides containing either RGD or PHSRN bioactive sequences were mixed and covalently bonded on CPTES-silanized surfaces. Coatings made of only one of the two investigated peptides and coatings with physisorbed oligopeptides were produced and tested as control groups. We performed thorough characterization of the obtained surfaces after each step of the coating preparation and after mechanically challenging the obtained coatings. Fluorescence labeling of RGD and PHSRN peptides with fluorescence probes of different colors enabled the direct visualization of the co-immobilization of the oligopeptides. We proved that the coatings were mechanically stable. The surfaces with co-immobilized RGD and PHSRN peptides significantly improved osteoblasts response in comparison with control surfaces, which assessed the effectiveness of our coating method to bio-activate the implant surfaces. This same simple method can be used to obtain other multi-functional surfaces by co-immobilizing oligopeptides with different targeted bioactivities--cell recruitment and differentiation, biomineral nucleation, antimicrobial activity--and thus, further improving the clinical performance of titanium implants.

Aging of Retrieved Zirconia Femoral Heads

Yttria-stabilized tetragonal zirconia may undergo extensive transformation to the monoclinic phase under mechanical and/or hydrothermal stress, with degradation of mechanical and tribologic properties. We hypothesized progressive phase transformation of zirconia in service in vivo is directly related to the time of implantation and to patient-related factors. The subsequent decrease in fracture toughness and increase in surface roughness and wear are related to the increased monoclinic content. We performed a study on 47 yttria-stabilized tetragonal zirconia femoral heads retrieved from failed total hip arthroplasties after 2 to 10 years implantation. Age, weight, and activity of the patients were retrieved from clinical records. Monoclinic content, fracture toughness, surface roughness, and wear were measured. Strong correlations were found between monoclinic content in the weightbearing surface and time of implantation (r = 0.97) and between increase in monoclinic content and decrease in fracture toughness (r = −0.92), increase in surface roughness (r = 0.88), and increase in surface wear (r = 0.89). No correlation was observed between the increase in monoclinic content and the age, weight, or activity of the patients. Aging of zirconia in vivo is then a function of time in service, and the loss of surface properties is caused by the corresponding increase in monoclinic content. Level of Evidence: Level II, prognostic study. See Guidelines for Authors for a complete description of levels of evidence.

Evaluation of ion release, cytotoxicity, and platelet adhesion of electrochemical anodized 316 L stainless steel cardiovascular stents

316L Stainless steel is one of the most used metallic material in orthopedical prosthesis, osteosinthesis plates, and cardiovascular stents. One of the main problems this material presents is the nickel and chromium release, specially the Ni ion release that provokes allergy in a high number of patients. Recently, experimental applications in vitro and in vivo seem to indicate that the thickness of the nature oxide of the stainless steel results in very strong reinforcement of the biological response and reduce the ion release due to the thicker surface oxide. It is possible to grow the natural chromium oxide layer by electrolytic method such anodization. In this study, two main anodization methods to grow chromium oxide on the 316L stainless steel have been evaluated. Nickel and Chromium ions release in human blood at 37 degrees C were detected at times of 1, 6, 11, and 15 days by means of atomic absorption in a graphite furnace (GAAF). Moreover, cytocompatibility tests were carried out. Perfusion experiments were performed to evaluate morphometrically platelet interaction with the material and to explore the potential thrombogenicity. The results showed a good cytocompatibility between the material and the osteoblast-like cells. However, these anodization methods released between 2 and 10 times more nickel and chromium than the original stainless steel, depending on the method used. Besides, anodized samples shown an increase of the percentage of surface covered by platelets. Consequently, the anodization methods studied do not improve the long-term behavior of the stainless steel for its application as cardiovascular stents.

In vitro evaluation of surface topographic changes and nickel release of lingual orthodontic archwires

The objective of the article is to study surface topographic changes and nickel release in lingual orthodontic archwires in vitro. Stainless steel (SS), nickel–titanium (NiTi) and copper–nickel–titanium (CuNiTi) lingual orthodontic archwires were studied using atomic absorption spectrometry for nickel release after immersion in a saline solution. Surface roughness changes were measured using atomic force microscopy. Differences between groups were analyzed using independent sample t-tests. Statistically significant changes in roughness were seen in all archwires except NiTi. Surface changes were most severe in the CuNiTi alloy. SS archwires released the highest amount of nickel. In conclusion, only roughness changes in CuNiTi archwires seemed to be clinically significant. The amount of nickel released for all archwires tested is below the levels known to cause cell damage.

Anhydride-functional silane immobilized onto titanium surfaces induces osteoblast cell differentiation and reduces bacterial adhesion and biofilm formation.

Bacterial infection in dental implants along with osseointegration failure usually leads to loss of the device. Bioactive molecules with antibacterial properties can be attached to titanium surfaces with anchoring molecules such as silanes, preventing biofilm formation and improving osseointegration. Properties of silanes as molecular binders have been thoroughly studied, but research on the biological effects of these coatings is scarce. The aim of the present study was to determine the in vitro cell response and antibacterial effects of triethoxysilypropyl succinic anhydride (TESPSA) silane anchored on titanium surfaces. X-ray photoelectron spectroscopy confirmed a successful silanization. The silanized surfaces showed no cytotoxic effects. Gene expression analyses of Sarcoma Osteogenic (SaOS-2) osteoblast-like cells cultured on TESPSA silanized surfaces reported a remarkable increase of biochemical markers related to induction of osteoblastic cell differentiation. A manifest decrease of bacterial adhesion and biofilm formation at early stages was observed on treated substrates, while favoring cell adhesion and spreading in bacteria-cell co-cultures. Surfaces treated with TESPSA could enhance a biological sealing on implant surfaces against bacteria colonization of underlying tissues. Furthermore, it can be an effective anchoring platform of biomolecules on titanium surfaces with improved osteoblastic differentiation and antibacterial properties.

Analysis of tantalum implants used for avascular necrosis of the femoral head: a review of five retrieved specimens

Aim The effective results shown in the porous systems of tantalum employed for the use of osseointegrates has been demonstrated by means of animal experimentation. However, there is a total lack of any research studies on the osseointegration of tantalum implants from retrieval of the same after a period of time whereby the material had been implanted within the human body. Materials and Methods For this study, five rod implants used for the treatment of avascular necrosis of the femoral head were retrieved following collapse of the femoral head and conversion to total hip arthroplasty. The time of implantation ranged between six weeks and twenty months. Results Observation during this study has confirmed the effectiveness of osseointegration within this period of time. New bone was observed around and within the porous system of the on rod devices at retrieval date. The bone ingrowth, however, proved to be slower and less intense than that resulting within animal species during the first few months after implantation. Conclusions The results obtained in the quantitative assessment of this process proved to be similar to those results achieved by other authors in previous experimental work studies.

Evaluation of bone loss in antibacterial coated dental implants: an experimental study in dogs

The aim of this study was to evaluate the in vivo effect of antibacterial modified dental implants in the first stages of peri-implantitis. Thirty dental implants were inserted in the mandibular premolar sites of 5 beagle dogs. Sites were randomly assigned to Ti (untreated implants, 10units), Ti_Ag (silver electrodeposition treatment, 10units), and Ti_TSP (silanization treatment, 10units). Coated implants were characterized by scanning electron microscopy, interferometry and X-ray photoelectron spectroscopy. Two months after implant insertion, experimental peri-implantitis was initiated by ligature placement. Ligatures were removed 2months later, and plaque formation was allowed for 2 additional months. Clinical and radiographic analyses were performed during the study. Implant-tissue samples were prepared for micro computed tomography, backscattered scanning electron microscopy, histomorphometric and histological analyses and ion release measurements. X-ray, SEM and histology images showed that vertical bone resorption in treated implants was lower than in the control group (P<0.05). This effect is likely due to the capacity of the treatments to reduce bacteria colonization on the implant surface. Histological analysis suggested an increase of peri-implant bone formation on silanized implants. However, the short post-ligature period was not enough to detect differences in clinical parameters among implant groups. Within the limits of this study, antibacterial surface treatments have a positive effect against bone resorption induced by peri-implantitis.

Biofunctionalization of titanium surfaces for osseintegration process improvement

This study aims to improve the osseointegration of titanium implants through surface immobilization of peptides that induce a beneficial biological response. This was carried out biofunctionalizating titanium surfaces by silanization and subsequent covalent binding of a peptide with a sequence that promotes cell adhesion. Objective: The development of a new technique of immobilization of oligopeptides on the surface of titanium by using 3-chloropropyltrietoxisilane (CPTES) as bonding agent between the surface of titanium and the peptide. Materials and methods: A physicochemical characterization of the surfaces through the techniques of XPS, ToF-SIMS and contact angle was performed. Also cell adhesion studies have been conducted to evaluate in vitro biological response. Results: Through the process of silanization the titanium surface is completely covered with CPTES, which allows the subsequent accession of oligopeptides. The cell adhesion results show a higher cell adhesion and cell extension on biofunctionalized samples. Conclusions: We developed a system of covalent binding of oligopeptides on titanium surfaces that can modify the biological response of the attached cells.

Surface immobilization and bioactivity of TGF-β1 inhibitor peptides for bone implant applications

TGF-β1 is the most related cytokine with the production of fibrotic tissue. It plays an important role on the production of collagen by fibroblasts and other types of cells. The inhibition of this cytokine reduces fibrosis in various types of tissue. Biofunctionalization of dental and orthopedic implants with biomolecules enables modification of the physical, chemical and biochemical properties of their surfaces to improve its biological and clinical performance. Our objective was to develop a reliable method to immobilize oligopeptides on Ti surfaces to obtain a surface with TGF-β1 inhibitory activity that will potentially minimize fibrotic encapsulation of implants during the process of osseointegration. We covalently immobilized TGF-β1 inhibitor P17-peptides on Ti surfaces and assessed by characterizing each step of the process that we successfully biofunctionalized the implant surfaces. High amounts of peptides were anchored and homogeneously distributed on the surfaces with mechanical and thermochemical stability after in vitro simulated challenges. Notably, the immobilized peptides retained their TGF-β1 inhibitory activity in vitro. Thus, these biofunctional coatings are potential candidates for inducing a fast and reliable osseointegration in vivo.

Fatigue Life of Bioactive Titanium Dental Implants Treated by Means of Grit-Blasting and Thermo-Chemical Treatment

OBJECTIVE This study focuses on the fatigue behavior of titanium dental implants as-received, with a grit-blasted surface and with a new bioactive surface treatment (2Steps). BACKGROUND The 2Step process consists of (1) an initial grit-blasting process to produce a micro-rough surface, followed by (2) a combined thermo-chemical treatment that produces a potentially bioactive surface, that is, that can form an apatitic layer when exposed to biomimetic conditions in vitro. The 2Step treatment produced micro-rough and apatitic coating implants. METHODS Residual stresses were determined by means of X-ray diffraction. The fatigue tests were carried out at 37°C on 500 dental implants, and the S-N curve was determined. The fatigue-crack nucleation for the different treatments was analyzed. RESULTS The fatigue tests show that the grit-blasting process improves the fatigue life. This is a consequence of the layer of compressive residual stresses that the treatment generates in titanium surfaces. Dental implants that had its surfaced prepared with the 2Step procedure (grit-blasting and thermo-chemical treatment) had its fatigue life decreased by 10% due to the incorporation of oxygen to the surface and the relaxation of the compressive residual stress produced by the heat treatment. CONCLUSIONS Thermo-chemical treatment is an excellent compromise between the improvement of bioactive and mechanical long-life behaviors.