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Effects of laser-modified versus osteopromotively coated titanium membranes on bone healing: a pilot study in rat mandibular defects.

Regeneration of alveolar bone with membrane techniques has become an integral part of implant dentistry. The aim of the present study was to determine if laser-modified titanium membranes are of value in the regeneration of so-called critical size defects in the rat model compared with titanium membranes that were coated with growth factors. A total of 24 rats were included in the study. Critical size defects were created bilaterally and covered by titanium membranes coated with (1). polylactide, (2). polylactide and clindamycin, (3). polylactide and growth factors, (4). polylactide, clindamycin and growth factors and (5). uncoated but laser-modified titanium membranes. All 18 control defects were covered by titanium membranes without any substrate. Four weeks after treatment the animals were killed. Laser-modified titanium membranes (group 5) showed new bone formation in many areas. Nevertheless, complete bridging was found only in one specimen. In contrast, in groups 3 and 4, most defects showed almost complete bridging of the defects. In particular, clindamycin had no inhibitory effect on bone healing. Furthermore, after 28 days, there was no significant difference between the individual groups (including controls) with respect to the total amount of lamellar bone. Growth-factor-coated membranes can significantly accelerate the healing process of bony defects in the rat mandibular model. Nevertheless, it is not possible to accelerate bone healing with laser-irradiated membranes or to enhance the quality of bone within the time period examined.

Animals↗

Improved mechanical properties of acrylic bone cement with short titanium fiber reinforcement.

Acrylic bone cements are widely used in total joint arthroplasties to grout the prosthesis to bone. The changes in the tensile properties and fracture toughness of polymethylmethacrylate (PMMA) bone cements obtained by the addition of control and heat treated short titanium fibers are studied. Heat treatment of titanium fibers is conducted to precipitate titania particles on the fiber surface, which may improve the biocompatibility of the metal. Control (non-heat treated) and heat treated short titanium fibers (250 microm long and 20microm diameter) were used as reinforcements at 3 volume %. X-ray diffraction indicated the presence of a rutile form of titania due to the heat treatments. Results indicate that the tensile and fracture properties of unfilled bone cement were improved by the addition of control and heat-treated fibers. The fracture properties of bone cements reinforced with control titanium fibers were at least 10% higher than those reinforced with heat treated titanium fibers. Therefore, we recommend further studies on the use of non-heat treated titanium fibers to reinforce acrylic bone cement.

Coated Materials, Biocompatible↗

Titanium alloy cylinders in implant framework fabrication: a study of the cylinder-alloy interface.

Titanium alloy has been proposed as a suitable alternative to noble metal alloys in premanufactured metal cylinders for fabricating implant prosthesis frameworks. The interfaces produced by standard lost wax procedures that use titanium and conventional noble metal implant cylinders with both high-fusing (high-palladium) and low-fusing (high-gold) noble metal casting alloys were compared. Elemental analysis by scanning electron microscopy and energy-dispersive spectroscopy (SEM/EDS) of the two titanium cylinders revealed that one commercial product had a composition similar to that for commercially pure titanium and the other had a composition consistent with that for Ti-6Al-4V alloy. The SEM observations and complementary elemental line scans indicated that the interfaces between the titanium cylinders and the two noble metal casting alloys did not meet proposed criteria for acceptable metal-to-metal unions. The interfaces between the noble metal cylinders and noble metal casting alloys did exhibit acceptable unions. The SEM observations and elemental analyses were supported by push-shear test results that showed that the resistance to failure was appreciably greater for interface specimens produced with alloy cast to noble metal cylinders compared with titanium cylinders.

Alloys↗

Effect of titanium implants on primary mineralization following 6 and 14 days of rat tibial healing.

The effect of pure commercial titanium implants on the process of primary mineralization was studied. This was examined by insertion of titanium implants into rat tibial bone after ablation. The effects of the titanium were studied through the behaviour of extracellular matrix vesicles (MV). Methods of morphometric analysis at the TEM level were applied. The insertion of titanium implants was followed by an increase in the number of MV as well as vesicular diameter and by a decrease in vesicular distance from the calcified front when compared to normal healing. These results suggest that the process of MV maturation around titanium implants was delayed when compared to normal primary bone formation during bone healing. The delay in mineralization was compensated by an increase in vesicle production, resulting in an enhancement of primary mineralization by the titanium.

Animals↗

Behaviour of titanium and titania-based ceramics in vitro and in vivo.

Cylinders, 2.8 mm in diameter and 6 mm in length which were produced from hot isostatically pressed titania (TI), titania-hydroxyapatite composite (TI/HA) and hydroxyapatite (HA) as well as commercially pure titanium (c.p. Ti), were implanted in the femurs of New Zealand white rabbits. The animals were killed at 1 and 3 months after implantation to study the development of bonding strength between the bone and the implant using the push-out test. The surface roughness and ion release of these materials in different solutions were also studied. Titania and titanium showed similar push-out strengths at 1 and 3 months after implantation. This indicates that bone has a similar response to titanium and titania due to the similarity of the surface constitute. Titania-hydroxyapatite composite showed a significant increase in bonding strength at 1 and 3 months after implantation compared with titania. A gap, < 2 microns wide, was observed by SEM at the bone-TI interface while the bone-HA interface was fully mineralized. No release of titanium ions from titania (rutile) was detected in the solutions used, including 0.1 M HCl, whilst release of titanium ions from c.p. Ti was measured in 0.02 M lactic acid and 0.1 M HCl. The results suggested that a further oxidation of titanium may improve the stability of c.p. Ti and that titania-based material may be a strong biocompatible ceramic suitable for medical applications.

Animals↗

Titanium, chromium and cobalt ions modulate the release of bone-associated cytokines by human monocytes/macrophages in vitro.

Osteolysis has become a major cause of aseptic loosening in total joint arthroplasty (TJA). Titanium, cobalt and chromium are commonly used in orthopaedic implants (e.g. joint prostheses). The release of bone-associated cytokines has been associated with the development of osteolysis in patients with prostheses. We evaluated the effects of these metals on the release of bone-associated cytokines (IL-1 beta, IL-6, TNF-alpha and TGF-beta 1) by human blood monocytes/macrophages and monocyte-like U937 cells upon lipopolysaccharide (LPS) stimulation, the cell proliferation, and their cytotoxic effects on these cells in vitro. We found that the release of IL-1 beta was enhanced by titanium, chromium and cobalt, the release of TNF-alpha was enhanced by titanium and chromium, and the release of IL-6 was enhanced by titanium. All three metal ions inhibited the release of TGF-beta 1. We also found that titanium and chromium, but not cobalt, enhanced blood monocyte/macrophage proliferation in response to LPS while only titanium enhanced U937 cell proliferation in response to LPS. The metals in concentrations ranging from 0.01 to 100 ngml-1 did not stimulate the cells to secrete detectable cytokines in the absence of LPS. Furthermore, a 4-h pre-exposure of blood monocytes/macrophages or U937 cells to the metals did not alter cytokine release when the metals were removed from the media prior to the addition of LPS. Similarly, a 4-h pre-exposure of blood monocytes/macrophages or U937 cells to LPS did not alter cytokine release when LPS was removed from the media prior to the addition of the metals. The metals did not reduce cell viability and induce cell injury after 72h incubation with the cells. The data suggest that the three metals at clinically relevant concentrations modulated cytokine expression, whereas they did not induce any cytotoxic effects. A metal-induced enhancement of bone-resorbing cytokine release with a concomitant inhibition of bone-forming cytokine release may be an important factor in the development of osteolysis, which can severely compromise the outcome of TJA.

Biocompatible Materials↗

Effects of sandblasting and silica-coating procedures on pure titanium.

Silica coating titanium improves chemomechanical bonding. Sandblasting is recommended as a pretreatment to thermal silica coating (Silicoater MD) or as part of a tribochemical silica coating process (Rocatec). This study evaluated the effects of sandblasting and coating techniques on volume loss, surface morphology and composition changes in pure titanium. Volume loss of titanium was similar to values reported for base alloys and does not seem to be critical for the clinical fit of restorations. Embedded alumina particles were found in the titanium after sandblasting and the alumina content increased to a range of 27.5-39.3 wt% as measured by EDS. Following tribochemical silica coating, a layer of small silica particles remained on the surface, increasing the silica content to a range of 17.9-19.5 wt%. Ultrasonic cleaning removed loose alumina or silica particles from the surface, resulting in only slight decreases in alumina or silica contents, suggesting firm attachment of most of the alumina and silica to the titanium surface. Silica content following thermal silica coating treatment increased only slightly from the sandblasted specimen to 1.4 wt%. The silica layer employed by these silica coating methods differs widely in both morphology and thickness. These results provide a basis for explanation of adhesive failure modes in bond strength tests and for developing methods to optimize resin bonding. Clinically, ultrasonic cleaning of sandblasted and tribochemically silica coated titanium should improve resin bonding as loose surface particles are removed without relevant changes in composition.

Aluminum Oxide↗

In-vitro evaluation of the material characteristics of stainless steel and beta-titanium orthodontic wires.

INTRODUCTION: The exact composition and material properties of the metal alloys used in orthodontics are usually not identified by or even available from manufacturers. This makes meaningful comparisons between wires impossible and is unacceptable with regard to biocompatibility issues. The aim of this study was to investigate the material characteristics of contemporary stainless steel (SS) and beta-titanium (beta-Ti) wires, also known as titanium-molybdenum alloy (TMA), for comparison. METHODS: Twenty-two different SS and beta-Ti wires, preferably straight wires sized 0.43 x 0.64 mm, (0.017 x 0.025 in) were tested blindly for wire dimensions, chemical compositions, bending and tensile properties, and surface characteristics. RESULTS: Four chemical compositions were found for the beta-Ti wires: titanium-11.5, molybdenum-6, zirconium-4.5 tin; titanium-3, aluminum-8, vanadium-6, chromium-4, molybdenum-4, zirconium; titanium-6, aluminum-4, vanadium, and titanium-45 niobium. The SS wires were of AISI type 304 or the nickel-free variant BioDur 108. All beta-Ti wires showed high surface roughness values. TMA 02 significantly had the highest E-modulus, TMA 02 and TMA 11 had the highest 0.2% yield strength, TMA 02 had the highest hardness, and TMA 12 was the most ductile wire of the beta-Ti wires. All SS wires showed high 0.2% yield strength, SS 10 significantly had the lowest E-modulus and was the most ductile wire, and SS 08 significantly showed the lowest hardness values of all SS wires. CONCLUSIONS: Significant differences were found between SS and beta-Ti wires, but there was little or no difference between the mechanical and physical characteristics tested in each subgroup. However, the morphological analysis clearly demonstrated that the finishing phase (annealing, polishing) of the wires' production process lacks the quality one would expect with regard to good mechanical properties and biocompatibility issues. Accurate specifications are urgently needed concerning the quality of orthodontic wires on the market.

Analysis of Variance↗

Biomimetic apatite formation on chemically treated titanium.

Titanium treated in NaOH can form hydroxycarbonated apatite (HCA) after exposition in simulated body fluid (SBF). Generally, titanium is covered with a passive oxide layer. In NaOH this passive film dissolves and an amorphous layer containing alkali ions is formed on the surface. When exposed to SBF, the alkali ions are released from the amorphous layer and hydronium ions enter into the surface layer, resulting in the formation of Ti-OH groups in the surface. The released Na(+) ions increase the degree of supersaturation of the soaking solution with respect to apatite by increasing pH, and Ti-OH groups induce apatite nucleation on the titanium surface. The acid etching of titanium in HCl under inert atmosphere was examined as a pretreatment to obtain a uniform initial titanium surface before alkali treatment. Acid etching in HCl leads to the formation of a micro-roughened surface, which remains after alkali treatment in NaOH. It was shown by SEM, gravimetric and solution analysis that the apatite nucleation was uniform and the thickness of precipitated HCA layer increased continuously with time. The treatment of titanium by acid etching in HCl and subsequently in NaOH is a suitable method for providing the metal implant with bone-bonding ability.

Apatites↗

Glow discharge plasma pretreatment enhances osteoclast differentiation and survival on titanium plates.

Despite the fact that several reports have demonstrated osteoclast activity on various bioactive ceramics, osteoclast functions on surface-modified titanium have not come under focus. This study aimed to examine whether the increasing surface energy of glow discharge plasma (GDP) involved in protein adhesion containing the RGD (Arg-Gly-Asp) sequence affects osteoclast responses on titanium plates. We examined osteoclast differentiation and survival rates on titanium plates with and without GDP. The amounts of osteoclasts on titanium plates were not increased by GDP after 1 week. However, osteoclast differentiation was greatly activated by GDP pretreatment, as tartrate-resistant acid phosphatase synthesis significantly increased on the titanium plates with GDP. Additionally, since the presence of osteoclasts was detected only on the titanium plates with GDP, even after 4h cultivation in a coculture test, the osteoclasts survival rate was increased by GDP pretreatment. As osteoclast responses were affected even on surface modified metallic materials, we concluded that novel approaches are needed not only for osteoclastic resorption on ceramic materials but also for osteoclast responses on surface-modified metallic materials.

Acid Phosphatase↗

Nano-scale study of the nucleation and growth of calcium phosphate coating on titanium implants.

The nucleation and growth of a calcium phosphate (Ca-P) coating deposited on titanium implants from simulated body fluid was investigated by using atomic force microscopy (AFM) and environmental scanning electron microscopy (ESEM). Forty titanium alloy plates were assigned into two groups. One group with a smooth surface having a maximum roughness R(max) < 0.10 microm (s-Ti6Al4V) and a group with a rough surface with an R(max) < 0.25 microm (r-Ti6Al4V) were used. Titanium samples were immersed in SBF concentrated by five (SBF x 5) from 10 min to 5 h and examined by AFM and ESEM. Scattered Ca-P deposits of approximately 15 nm in diameter appeared after only 10 min of immersion in SBF x 5. These Ca-P deposits grew up to 60-100 nm after 4 h on both s- and r-Ti6Al4V substrates. With increasing immersion time, the packing of Ca-P deposits with size of tens of nanometers in diameter formed larger globules and then a continuous Ca-P film on titanium substrates. A direct contact between the Ca-P coating and the Ti6Al4V surface was observed. The Ca-P coating was composed of nanosized deposits and of an interfacial glassy matrix. This interfacial glassy matrix might ensure the adhesion between the Ca-P coating and the Ti6Al4V substrate. In the case of s-Ti6Al4V substrate, failures within this interfacial glassy matrix were observed overtime. Part of the glassy matrix remained on s-Ti6Al4V while part detached with the Ca-P film. The Ca-P coating detached from the smooth substrate, whereas the Ca-P film extended onto the whole rough titanium surface over time. In the case of r-Ti6Al4V, the Ca-P coating covered evenly the substrate after immersion in SBF x 5 for 5 h. The present study suggested that the heterogeneous nucleation of Ca-P on titanium was immediate and did not depend on the Ti6Al4V surface topography. The further growth and mechanical attachment of the final Ca-P coating strongly depended on the surface, for which a rough topography was beneficial.

Alloys↗

Platelet adhesion on titanium oxide gels: effect of surface oxidation.

The correlations between titanium oxide layers on oxidized titanium (Ti) substrates and platelet adhesion were examined. Ti substrates were prepared by three different oxidation methods: the first one was treated with hydrogen peroxide (H2O2) solution, the second one was heated in air at moderate temperatures, and the third one was processed with both H2O2 and heating. The titanium oxide layers formed on the Ti substrates were characterized by wettability, chemical composition, thickness, and crystal phase. The platelet adhesion on these oxide layers was examined and correlated to the characterizations of the surface layers. The number of adhesive platelets seemingly correlated with the contact angle towards distilled water, because the number increased close to 70-80 degrees of the contact angle. The effect of surface oxidation on platelet adhesion was examined in detail and it was found that the composition and thickness of the oxide layer influenced platelet adhesion rather than wettability. Thick titanium oxide layers formed on Ti substrates by heating displayed less platelet adhesion than thin oxide layers on untreated Ti substrates. The largest number of adhesive platelets was found on H2O2-oxidized substrates; the substrates found on amorphous titanium oxide contained the Ti-peroxide radical species. The number of platelets, on the other hand, could hardly be observed on Ti substrates treated with H2O2 and subsequently heated above 300 degrees C. The titanium oxide layer on the substrate was thick and we found it to consist of only a few radical species. That is, the effect of heat treatment accelerates the growth of the oxide layer, and decomposes or decreases the number of radical species. Ti substrates with H2O2 and heat treatment above 300 degrees C held the least number of platelets, and were concluded to be the most inhibitory for platelet adhesion.

Blood Platelets↗

Biocompatibility of beta-stabilizing elements of titanium alloys.

In comparison to the presently used alpha + beta titanium alloys for biomedical applications, beta-titanium alloys have many advantageous mechanical properties, such as an improved wear resistance, a high elasticity and an excellent cold and hot formability. This will promote their future increased application as materials for orthopaedic joint replacements. Not all elements with beta-stabilizing properties in titanium alloys are suitable for biomaterial applications-corrosion and wear processes cause a release of these alloying elements to the surrounding tissue. In this investigation, the biocompability of alloying elements for beta- and near beta-titanium alloys was tested in order to estimate their suitability for biomaterial components. Titanium (grade 2) and the implant steel X2CrNiMo18153 (AISI 316 L) were tested as reference materials. The investigation included the corrosion properties of the elements, proliferation, mitochondrial activity, cell morphology and the size of MC3T3-E1 cells and GM7373 cells after 7 days incubation in direct contact with polished slices of the metals. The statistical significance was considered by Weir-test and Lord-test (alpha = 0.05). The biocompatibility range of the investigated metals is (decreasing biocompatibility): niobium-tantalum, titanium, zirconium-aluminium-316 L-molybdenum.

3T3 Cells↗

The bisphosphonate pamidronate on the surface of titanium stimulates bone formation around tibial implants in rats.

Many materials with differing surfaces have been developed for clinical implant therapy in dentistry and orthopedics. We analyzed the quantity of new bone formed in vivo around calcium-immobilized titanium implants with surfaces modified using pamidronate (PAM), a nitrogen-containing bisphosphonate (N-BP), implants of pure titanium, and titanium implants immobilized with calcium ions. New bone formation was visualized using fluorescent labeling (calcein blue and alizarin complexone) with intravenous injection at 1 and 3 weeks after implantation. After 4 weeks, undecalcified sections were prepared, and new bone formation around the implants was examined by morphometry using confocal laser scanning microscopy images. After 1 week, more new bone formed around the PAM-immobilized implant than around the calcium-immobilized and pure titanium implants. This was also seen with the new bone formation after 3 weeks. After 4 weeks, significantly more new bones were formed around the BP-immobilized implant than around the calcium ion-implanted and pure titanium implants. The new N-BP-modified titanium surface stimulates new bone formation around the implant, which might contribute to the success of implant therapy.

Animals↗

A comparative study of electrochemical deposition and biomimetic deposition of calcium phosphate on porous titanium.

Coating porous titanium with calcium phosphate (Ca-P) is an effective way to enhance titanium's osteoinduction capability. This study investigated the effectiveness of two coating methods: biomimetic deposition (BD) and electrochemical deposition (ED) in aqueous solutions. The titanium surfaces were treated by acidic etching and alkaline before coating. Effects of the pre-coating treatments on Ca-P coating were also investigated. Both deposition methods could produce Ca-P coatings on the inner pore surfaces of the titanium. The BD coatings were thicker and more uniform than were the ED coatings. On the other hand, ED was less sensitive to the condition of the titanium surface, and much faster in the coating deposition. However, ED produces less uniform and thinner coating layers on the inner pore surfaces of the titanium than does BD. The crystal structure of the coating is octacalcium phosphate (OCP) regardless of the deposition method. The morphology of flake-like OCP crystals in the deposition layers is similar for both deposition methods, except that the crystal flakes rupture after ED.

Biocompatible Materials↗

Improvement of surface bioactivity on titanium by water and hydrogen plasma immersion ion implantation.

We have investigated the surface bioactivity of titanium after water and hydrogen plasma immersion ion implantation. Plasma immersion ion implantation (PIII) excels in the surface treatment of components possessing a complicated shape such as medical implants. In addition, water and hydrogen PIII has been extensively studied as a method to fabricate silicon-on-insulator (SOI) substrates in the semiconductor industry and so it is relatively straightforward to transfer the technology to the biomedical field. In our investigation, water and hydrogen were plasma-implanted into titanium sequentially. Our objective is that water PIII introduces near-surface damages that trap hydrogen implanted in the subsequent step to improve the surface bioactivity while the desirable bulk properties of the materials are not compromised. Ti-OH functional groups can be detected on the (H(2)O+H(2))-implanted titanium surface by X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) spectroscopy. After incubation in simulated body fluids (SBF) for cytocompatibililty evaluation in vitro, bone-like hydroxyapatite was found to precipitate on the (H(2)O+H(2)) implanted samples while no apatite was found on titanium samples plasma implanted with water or hydrogen alone. Human osteoblast cells were cultured on the (H(2)O+H(2))-implanted titanium surface and they exhibited good adhesion and growth. Our results suggest a practical means to improve the surface bioactivity and cytocompatibility of medical implants made of titanium.

Biocompatible Materials↗

Osteoinductive porous titanium implants: effect of sodium removal by dilute HCl treatment.

In a previous study, we observed that chemically and thermally treated plasma-sprayed porous titanium possesses intrinsic osteoinductivity and that bone formation occurs after 12 months in the muscles of beagle dogs. The aim of this study was to optimize the surface treatment and to accelerate the osteoinductivity. Previous studies have reported that sodium removal converts the sodium titanate layer on the surface of an alkali-treated titanium plate into a more bioactive titania layer. In this study, we developed a dilute hydrochloric acid (HCl) treatment for porous titanium, which removed sodium from the complexly shaped porous structure more effectively than conventional hot water treatment. Three types of surface treatments were applied: (a) alkali and heat treatment (AH treatment); (b) alkali, hot water, and heat treatment (Water-AH treatment); and (c) alkali, dilute HCl, hot water, and heat treatment (HCl-AH treatment). The osteoinductivity of the materials implanted in the back muscles of adult beagle dogs was examined at 3, 6, and 12 months. The HCl-AH-treated porous bioactive titanium implant had the highest osteoinductivity, with induction of a large amount of bone formation within 3 months. The dilute HCl treatment was considered to give both chemical (titania formation and sodium removal) and topographic (etching) effects on the titanium surface, although we cannot determine which is the predominant factor. Nevertheless, adding the dilute HCl treatment to the conventional chemical and thermal treatments is a promising candidate for advanced surface treatment of porous titanium implants.

Animals↗

The role of whole blood in thrombin generation in contact with various titanium surfaces.

Understanding of the thrombotic response (activation of the intrinsic coagulation system followed by platelet activation) from blood components upon contact with a titanium dental implant is important and not fully understood. The aims of this study were to evaluate: (1) the thrombogenic response of whole blood, platelet-rich plasma (PRP) and platelet-poor plasma (PPP) in contact with a highly thrombogenic surface as titanium, (2) the thrombogenic response of clinically used surfaces as hydroxyapatite (HA), machined titanium (mTi), TiO2 grit-blasted titanium (TiOB) and fluoride ion-modified grit-blasted titanium (TiOB-F). An in vitro slide chamber model, furnished with heparin, was used in which whole blood, PRP or PPP came in contact with slides of the test surfaces. After incubation (60 min rotation at 22 rpm in a 37 degrees C water bath), blood/plasma was mixed with EDTA or citrate, further centrifuged at +4 degrees C (2200 g at 10 min). Finally, plasma was collected pending analysis. Whole blood in contact with Ti alloy resulted in the binding of platelets to the material surface and in the generation of thrombin-antithrombin (TAT) complexes. With whole blood TAT levels increased 1000-fold compared with PRP and PPP, in which both almost no increase of TAT could be detected. In addition, the platelet activation showed a similar pattern with a 15-fold higher release of beta-TG in whole blood. In the in vitro chamber model with the clinically relevant materials, the fluoride-modified surface (TiOB-F) showed pronounced TAT generation compared with TiOB, mTi and HA. Similar results were achieved for platelet consumption and activation markers of the intrinsic coagulation system. Taken together these results implicate first that whole blood is necessary for sufficient thrombin generation and platelet activation during placement of implants. Second, a fluoride ion modification seems to augment the thrombogenic properties of titanium.

Biocompatible Materials↗