Biomechanical and histomorphometric evaluation of skin integration on titanium and PEEK implants with different surface treatments.

Percutaneous implants are frequently affected by bacterial growth at the skin-implant interface. Integration between implant and surrounding skin is important to prevent bacteria from spreading to the underlying tissue. The standard method to evaluate skin-implant integration is by histomorphometry on samples which have been placed in tissue grown in vivo or ex vivo. In this study, a biomechanical method was developed and evaluated. The integration of implants into porcine skin was studied in an ex vivo model, where pig skin samples were cultivated in a nutrient solution. Cylindrical shaped implants, consisting of polyether ether ketone (PEEK) and titanium (Ti) with different surface treatments, were implanted in the skin tissue and the skin was grown in nutrient solution for 2 weeks. The implants were then extracted from the implantation site and the mechanical force during extraction was measured as a quantitative assessment of skin-implant integration. Implants from each group were also processed for histomorphometry and the degree of epidermal downgrowth (ED) and tissue to implant contact (TIC) was measured. A higher mean pullout force was observed for the PEEK implants compared to the Ti implants. Applying nanosized hydroxyapatite (HA) on Ti and PEEK increased the pullout force compared to uncoated controls, 24% for machined and 70% for blasted Ti, and 51% for machined PEEK. Treatment of Ti and PEEK with nanosized zirconium phosphate (ZrP) did not increase the pullout force. The histomorphometry analysis showed correlation between ED and pullout force, where the pullout force was inversely proportional to ED. For TIC, no significant differences were observed between the groups of same material (i.e. Ti, Ti+HA, Ti+ZrP, and PEEK, PEEK + HA, PEEK + ZrP), but it was significantly higher for PEEK compared to Ti. Scanning electron microscopy analysis was done on samples before and after the pullout tests, showing that the ZrP coating was unaffected by the 2 week ex vivo implantation and pullout procedure, no dissolution or detachment of the coating was observed. For the HA coating, a loss of coating was seen on approximately 5% of the total surface area of the implant. Graphical abstract.

Review of PEEK implants and biomechanical and immunological responses to a zirconium phosphate nano-coated PEEK, a blasted PEEK, and a turned titanium implant surface.

PURPOSE: To investigate the biomechanical and immunological reactions to coated and non-coated blasted PEEK implants in vivo after 12 weeks and review the associated literature. METHODS: Two osteotomy sites were performed in each proximal tibia of 10 lop-eared rabbits (n= 4 per rabbit). Each rabbit received a randomly placed (1) blasted zirconium phosphate nano-coated PEEK- (nano-ZrP), (2) blasted PEEK- (PEEK) and (3) titanium implant (Ti) and an empty sham site. At 12 weeks, removal torque of all implants and biological investigation with qPCR was performed. The implant surfaces were analyzed prior to insertion with interferometry, SEM and XPS. RESULTS: The interferometry analysis showed that there was no difference in roughness for the uncoated PEEK compared to the ZrP coated PEEK implants. The titanium implants were considerably smoother (Sa= 0.23 µm) than the uncoated Sa= 1.11 µm) and ZrP coated PEEK implants (Sa= 1.12 µm). SEM analysis on the PEEK implants corroborated the interferometry results; no difference in structure between the uncoated vs. the ZrP coated PEEK was visible on the micrometer level. At higher magnifications, the ZrP coating was visible in the SEM as a thin, porous network. All tested implants displayed osseointegration with the highest RTQ for nano-ZrP (18.4 Ncm) followed by PEEK (14.5 Ncm) and Ti (11.5 Ncm). All implants activated the immune system, with elevated macrophage and M2 macrophage qPCR markers at 12 weeks compared to the sham site. CLINICAL SIGNIFICANCE: Nano-ZrP coating improves osseointegration of blasted PEEK implants at 12 weeks of follow-up. Osseointegration of titanium, PEEK and nano-ZrP PEEK is not a normal bone healing process, but rather a shield-off mechanism that appears to be regulated by the innate immune system.

Evaluation of Implant Surface Modification with Nanohydroxyapatite Associated with the Use of L-PRF: In Vivo Study in Rats.

Leukocyte-platelet-rich fibrin (L-PRF) contains growth factors that stimulate bone regeneration. This study evaluated the bone repair in a tibia rat model around two implant surfaces in combination or not with L-PRF by assessing microtomographic and histomorphometric parameters. A total of 48 female rats were used in the study, in which 24 received implants with two types of surface treatments (dual acid etched-DAE or nanohydroxyapatite-nanoHA), and the other 24 received the same mini implants with L-PRF, which was collected by cardiac puncture, centrifugated, and inserted in the bone bed. The animals were euthanized 7 and 30 days after implant placement, and the retrieved samples were prepared for microtomographic and histomorphometric (bone-to-implant contact-BIC; and Bone Area Fraction Occupancy-BAFO) analyses. The adhesion of the nanoHA surface onto the implant surface was investigated by insertion and removal in simulated bone medium (Sawbones). The adhesion evaluation revealed that the loss of nanoHA after this procedure (as measured with SEM) from the implant surface was less than 1%. Overall, the nanoHA surface presented more bone in contact and in proximity to the implant, a higher bone surface/tissue volume fraction, a higher number of bone trabeculae, as well as trabecular separation relative to the DAE surface. Such results were more evident when the nanoHA surface was combined with L-PRF and after 30 days in vivo. The nanoHA surface presented higher BAFO when compared to DAE, with or without association with L-PRF. Therefore, implants with a nanoHA surface potentially benefit from the association to L-PRF.

Effect of Smoke Exposure on Gene Expression in Bone Healing around Implants Coated with Nanohydroxyapatite.

This study evaluated the effect of smoke exposure on the expression of genes related to bone metabolism in implants coated with nanohydroxyapatite (NHA). A total of 36 rats were exposed to cigarette smoke for 60 days. The animals were allocated into three groups: machined implants (MAC), dual acid-etched implants (DAE), and NHA-coated implants (NHA). Implants were installed in the left tibia of the rats after 30 days of smoke exposure. The implants were retrieved 7 and 30 days after implantation, and the adjacent bone analyzed using a real-time polymerase chain reaction for gene expression of alkaline phosphatase (ALP), osteopontin (OPN), receptor activator of the nuclear factor kappa ligand (RANKL), osteoprotegerin (OPG), the RANKL/OPG ratio, osteocalcin (OCN) and runt-related transcription factor 2 (Runx2). After 7 days, Runx2, OPN and OPG expression demonstrated significantly higher levels for the NHA surface treatment relative to DAE and MAC surfaces. NHA presented the lowest RANKL and RANKL/OPG levels. After 30 days, NHA-coated implants showed significantly higher levels of Runx2, ALP, OPN, OPG, OC, RANKL and RANKL/OPG relative to DAE and MAC implants. The results indicated a greater osteogenic and high osteoclastic activity around NHA implants, in comparison to DAE and MAC implants.

Nucleation and growth of calcium phosphates in the presence of fibrinogen on titanium implants with four potentially bioactive surface preparations. An in vitro study.

The aim of this study was to compare the nucleating and crystal growth behaviour of calcium phosphates on four types of potentially bioactive surfaces, using the simulated body fluid (SBF) model with added fibrinogen. Blasted titanium discs were modified by alkali and heat treatment, anodic oxidation, fluoride treatment, or hydroxyapatite coating. The discs were immersed in SBF with fibrinogen for periods of 3 days and 1, 2, 3 and 4 weeks. The topography, morphology, and chemistry of the surfaces were evaluated with optical interferometry, scanning electron microscopy/energy dispersive X-ray analysis (SEM/EDX), and x-ray photoelectron spectroscopy (XPS), respectively. All surface modifications showed early calcium phosphate formation after 3 days, and were almost completely covered by calcium phosphates after 2 weeks. After 4 weeks, the Ca/P ratio was approximately 2.0 for all surface groups except the fluoride modified surface, which had a Ca/P ratio of 1.0-1.5. XPS measurements of the nitrogen concentration, which can be interpreted as an indirect measure of the protein content, reached a peak value after 3 days immersion and decreased thereafter. In conclusion, the results in the present study, when compared to earlier SBF studies without proteins, showed that fibrinogen stimulates calcium phosphates formation. Furthermore, no pronounced differences could be detected between blasted controls and the potentially bioactive specimens.

Onyx Embolization for Occlusion of the Proximal Internal Iliac Artery During EVAR in Patients with Unsuitable Landing Zones in the Common Iliac Artery.

BACKGROUND: Twenty percent of the patients with AAA have an aneurysm involving the common iliac arteries. Large common iliac diameter can be treated with an iliac branched device or extension of the stent graft to the external iliac artery with occlusion of the ipsilateral internal iliac artery (IIA) to prevent type 2 endoleaks. This study describes and evaluates a embolization technique using Onyx in conjunction with EVAR in aneurysms with poor landing zones in the common iliac arteries. METHODS: Patients with Onyx IIA embolization during EVAR, identified from the hospital operating code database, constitute the study population. Onyx embolization was performed by injection at the IIA origin. Peri- and postoperative complications were collected from the medical records. Thin-sliced CT scan was performed 1 month and 1 year after the procedure. RESULTS: Thirty-six patients with complex iliac anatomy and insufficient landing zones (without sealing possibility for standard stent grafts) were identified out of 243 consecutive EVAR treatments during a 13-year period. In seventeen patients (7%), the IIA was embolized with Onyx. Technical success was obtained in all 17 patients, without adverse event or procedural complication. No complication related to the embolization procedure was noted during follow-up. CONCLUSIONS: During EVAR treatment of patients with aneurysm involving the common iliac artery, Onyx embolization of IIA is a feasible option without need of selective catheterization of the IIA orifice, potentially preserving important branches of the IIA and simplifying emergency procedures.

Biomechanical, histological, and computed X-ray tomographic analyses of hydroxyapatite coated PEEK implants in an extended healing model in rabbit.

A nanosized hydroxyapatite (HA) modification on polyetheretherketone (PEEK) using a novel spin coating technique was investigated in a rabbit model. Spin coating technique creates a 20-40 nm thick layer of nanosized HA particles with similar shape, size, and crystallinity as human bone. Some implants were designed with a perforating hole in the apical region to mimic a fusion chamber of a spinal implant. The coating nano-structures were assessed using a scanning electron microscope. The in vivo response to HA-PEEK was compared to untreated PEEK with respect to removal torque, histomorphometry, and computed microtomography. The HA-coated and pure PEEK implants were inserted in the tibia and femur bone according to simple randomization. The rabbits were sacrificed 20 weeks after implantation. Removal torque analysis showed significantly higher values for HA-PEEK. Qualitative histological evaluation revealed an intimate contact between PEEK and the bone at the threads and perforated hole. Histomorphometric assessment showed higher bone-implant and bone area values for HA-PEEK but without statistical significance. The effect of the HA coating showed most prominent effect in the removal torque which may be correlated to an alteration in the bone quality around the HA-PEEK implants. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1440-1447, 2018.

On the complexity of screening detected abdominal aortic aneurysms: a retrospective observational multicenter cohort study.

BACKGROUND: Ultrasound screening for abdominal aortic aneurysms (AAA) has been shown to decrease aneurysm related mortality. Likely by providing an opportunity to intervene while the aneurysm is still intact, but possibly also when and the anatomy still relatively uncomplicated which would provide a less complex procedure. Our aim was to retrospectively investigate the complexity of repair for screening-detected AAAs in a cohort of 65-year-old men. METHODS: All screening detected AAA cases that underwent repair between Sept 2010 and June 2014 in the most southern region of Sweden were included. Procedures were classified as either standard or complex. A standard procedure was defined as either standard EVAR (endovascular aneurysm repair) within the manufacturers Instructions For Use (IFU) or open repair with infrarenal clamping followed by a tube graft repair. All other types of procedures were defined as complex. The prevalence rate of AAA, screening compliance, short- and midterm outcome of the operations were reported. RESULTS: From the 35513 men invited to screening, 27 951 (78.7%) attended screening with ultrasound. AAA≥30 mm was found in 561 cases, yielding a prevalence rate of 2.0%. Forty-eight patients underwent AAA repair. A percentage of 43.8% of these were classified as complex procedures. These consisted mostly of branched/fenestrated EVAR or EVAR with simultaneous exclusion of common iliac aneurysm. CONCLUSIONS: Our study confirms contemporary prevalence rates of AAA. Almost half (43.8%) of screening-detected AAA required complex operations, a significant proportion. The complex aneurysms were, on average, larger than the non-complex cases and they were more likely to be cases that required surgery immediately after screening detection. Our data suggests that the nature of AAA is heterogenous, even in the screening-detected group requiring operation. This should spur interest in more studies to investigate this issue.

Polyether ether ketone implants achieve increased bone fusion when coated with nano-sized hydroxyapatite: a histomorphometric study in rabbit bone.

Polyether ether ketone (PEEK) possesses excellent mechanical properties similar to those of human bone and is considered the best alternative material other than titanium for orthopedic spine and trauma implants. However, the deficient osteogenic properties and the bioinertness of PEEK limit its fields of application. The aim of this study was to limit these drawbacks by coating the surface of PEEK with nano-scaled hydroxyapatite (HA) minerals. In the study, the biological response to PEEK, with and without HA coating, was investigated. Twenty-four screw-like and apically perforated implants in the rabbit femur were histologically evaluated at 3 weeks and 12 weeks after surgery. Twelve of the 24 implants were HA coated (test), and the remaining 12 served as uncoated PEEK controls. At 3 weeks and 12 weeks, the mean bone-implant contact was higher for test compared to control (P<0.05). The bone area inside the threads was comparable in the two groups, but the perforating hole showed more bone area for the HA-coated implants at both healing points (P<0.01). With these results, we conclude that nano-sized HA coating on PEEK implants significantly improved the osteogenic properties, and in a clinical situation this material composition may serve as an implant where a rapid bone fusion is essential.

Precipitation of Calcium Phosphates in the Presence of Collagen Type I on Four Different Bioactive Titanium Surfaces: an in Vitro Study.

OBJECTIVES: To compare the properties of calcium phosphate precipitation on four different bioactive surface preparations and one control surface in the simulated body fluid model with added collagen type I. MATERIAL AND METHODS: Blasted titanium discs were treated with four different surface modifications, alkali and heat, sodium fluoride, anodic oxidation and hydroxyapatite coating. The discs were divided into five groups where one group, the blasted, served as control. The discs were immersed in simulated body fluid and collagen for 24 h, 3 days, 1 week and 2 weeks and then analysed by optical interferometry, scanning electron microscopy/energy dispersive X-ray analysis and X-ray photoelectron spectroscopy. RESULTS: All surfaces show small precipitates after 3 days which with longer immersion times increase. After 2 weeks the surfaces were completely covered with precipitates, and Ca/P ratios were approximately 1.3, independently on surface preparation. The fluoridated discs showed significantly (P ≤ 0.05) higher degree of CaP after one week of immersion as compared to the other surface preparations. The collagen type I content increased with time, as reflected by increased nitrogen content. CONCLUSIONS: The results from this study indicate that a fluoridated titanium surface may favour precipitation of calcium phosphate in the presence of collagen type I, as compared to the other surface treatments of the present study.

Nanosized Hydroxyapatite Coating on PEEK Implants Enhances Early Bone Formation: A Histological and Three-Dimensional Investigation in Rabbit Bone.

Polyether ether ketone (PEEK) has been frequently used in spinal surgery with good clinical results. The material has a low elastic modulus and is radiolucent. However, in oral implantology PEEK has displayed inferior ability to osseointegrate compared to titanium materials. One idea to reinforce PEEK would be to coat it with hydroxyapatite (HA), a ceramic material of good biocompatibility. In the present study we analyzed HA-coated PEEK tibial implants via histology and radiography when following up at 3 and 12 weeks. Of the 48 implants, 24 were HA-coated PEEK screws (test) and another 24 implants served as uncoated PEEK controls. HA-coated PEEK implants were always osseointegrated. The total bone area (BA) was higher for test compared to control implants at 3 (p < 0.05) and 12 weeks (p < 0.05). Mean bone implant contact (BIC) percentage was significantly higher (p = 0.024) for the test compared to control implants at 3 weeks and higher without statistical significance at 12 weeks. The effect of HA-coating was concluded to be significant with respect to early bone formation, and HA-coated PEEK implants may represent a good material to serve as bone anchored clinical devices.

Biomechanical evaluation and surface characterization of a nano-modified surface on PEEK implants: a study in the rabbit tibia.

Polyether ether ketone (PEEK) is today frequently used as a biomaterial in different medical operations due to its excellent mechanical and chemical properties. However, the untreated surface of PEEK is bioinert and hydrophobic, and it does not osseointegrate in its pure form. The aim of this study was to evaluate a unique nano-modified surface of PEEK with respect to osseointegration. Forty-eight threaded, non-cutting PEEK implants were inserted bilaterally in the tibia of 24 rabbits. Half of the implants (n=24) were coated with nanocrystalline hydroxyapatite (test) and the remaining implants (n=24) were left uncoated (control). Half of the animals (n=12) were euthanized after 3 weeks of healing and the remaining (n=12) after 12 weeks. The implant retention was measured with a removal torque apparatus. Surface analysis was performed with interferometry, scanning electron microscopy, and X-ray photon spectroscopy to relate the removal torque to the applied surface. The test implants revealed a significantly higher retention after 3 weeks (P=0.05) and 12 weeks (P=0.028) compared to controls. The result of the present study proves that the addition of nanocrystalline hydroxyapatite coating to PEEK surfaces significantly increases its removal torque and biocompatibility.

Evaluation of bone healing on sandblasted and Acid etched implants coated with nanocrystalline hydroxyapatite: an in vivo study in rabbit femur.

This study aimed at investigating if a coating of hydroxyapatite nanocrystals would enhance bone healing over time in trabecular bone. Sandblasted and acid etched titanium implants with and without a submicron thick coat of hydroxyapatite nanocrystals (nano-HA) were implanted in rabbit femur with healing times of 2, 4, and 9 weeks. Removal torque analyses and histological evaluations were performed. The torque analysis did not show any significant differences between the implants at any healing time. The control implant showed a tendency of more newly formed bone after 4 weeks of healing and significantly higher bone area values after 9 weeks of healing. According to the results from this present study, both control and nano-HA surfaces were biocompatible and osteoconductive. A submicron thick coating of hydroxyapatite nanocrystals deposited onto blasted and acid etched screw shaped titanium implants did not enhance bone healing, as compared to blasted and etched control implants when placed in trabecular bone.

Nano-hydroxyapatite-coated PEEK implants: a pilot study in rabbit bone.

Osseointegration of surface-modified polyetheretherketone (PEEK) implants was studied in vivo. A total of 18 cylinder-shaped PEEK implants were inserted in the femurs of nine New Zealand rabbits; half were coated with nanocrystalline hydroxyapatite (nanoHA) and half were uncoated controls. Healing time was 6 weeks. Samples were retrieved with the implant and surrounding tissue, processed to cut and ground sections, and analyzed histomorphometrically. The implant surfaces were analyzed with optical interferometry, scanning electron microscopy (SEM), atomic force microscopy, and X-ray photoelectron spectroscopy (XPS). NanoHA-coated PEEK surfaces had lower height deviation (Sa) than controls [mean ± SD: 0.41 µm (± 0.14) vs. 0.96 µm (± 0.28)]. SEM images showed the nanoHA crystals as a thin layer on the polymer surface. XPS analysis of the coated implants showed a Ca/P ratio of 1.67. Histomorphometry indicated that the nanoHA-coated implants had more bone-to-implant contact [16% (± 4.7) vs. 13% (± 9.3)] and more bone area [52% (± 9.5) vs. 45% (± 11.9)]. We found no difference between smooth nanoHA-coated cylinder-shaped PEEK implants and uncoated controls. However, higher mean bone-to-implant contact indicated better osseointegration in the coated implants than in the uncoated controls. The large number of lost implants was interpreted as a lack of primary stability due to implant design.

Early interactions between leukocytes and three different potentially bioactive titanium surface modifications.

The aim of the present study was to compare the early interactions between leukocytes and three different surface modifications, suggested as bioactive. Blasted titanium discs were modified by alkali and heat treatment, sodium fluoride treatment, or hydroxyapatite coating. A number of these discs were also immersed in simulated body fluid (SBF) for a week, a treatment which yielded high levels of calcium and phosphate on each surface type. The specimens were exposed for human venous blood for 32 minutes and the respiratory burst response was measured in terms of reactive oxygen species with a luminometer, and coverage of viable cells with a fluorescence microscope after staining steps. The topography, morphology, and chemistry of the surfaces were evaluated with optical interferometry and scanning electron microscopy/energy dispersive X-ray analysis (SEM/EDX). A high respiratory burst response was found for HA coated titanium in comparison with the other surface groups (p < 0.0005). The SBF immersion resulted in an increased respiratory burst response (p < 0.0005) and removed statistically significant differences between the surface groups. Thus, the results in the present study indicate that different titanium surface modifications influence the early inflammatory response differently, and that calcium phosphate compounds increase the inflammatory response.

Bone reaction to nano hydroxyapatite modified titanium implants placed in a gap-healing model.

Nanohydroxyapatite materials show similar chemistry to the bone apatite and depending on the underlying topography and the method of preparation, the nanohydroxyapatite may simulate the specific arrangement of the crystals in bone. Hydroxyapatite (HA) and other CaP materials have been indicated in cases in which the optimal surgical fit is not achievable during surgery, and the HA surface properties may enhance bone filling of the defect area. In this study, very smooth electropolished titanium implants were used as substrata for nano-HA surface modification and as control. One of each implant (control and nano HA) was placed in the rabbit tibia in a surgical site 0.7 mm wider than the implant diameter, resulting in a gap of 0.35 mm on each implant side. Implant stability was ensured by a fixating plate fastened with two side screws. Topographical evaluation performed with an optical interferometer revealed the absence of microstructures on both implants and higher resolution evaluation with AFM showed similar nanoroughness parameters. Surface pores detected on the AFM measurements had similar diameter, depth, and surface porosity (%). Histological evaluation demonstrated similar bone formation for the nano HA and electropolished implants after 4 weeks of healing. These results do not support that nano-HA chemistry and nanotopography will enhance bone formation when placed in a gap-healing model. The very smooth surface may have prevented optimal activity of the material and future studies may evaluate the synergic effects of the surface chemistry, micro, and nanotopography, establishing the optimal parameters for each of them.

Effect of hydroxyapatite and titania nanostructures on early in vivo bone response.

PURPOSE: Hydroxyapatite (HA) or titania nanostructures were applied on smooth titanium implant cylinders. The aim was to investigate whether nano-HA may result in enhanced osseointegration compared to nano-titania structures. MATERIALS AND METHODS: Surface topography evaluation included detailed characterization of nano-size structures present at the implant surface combined with surface roughness parameters at the micro- and nanometer level of resolution. Microstructures were removed from the surface to ensure that bone response observed was dependent only on the nanotopography and/or chemistry of the surface. Early in vivo histological analyses of the bone response (4 weeks) were investigated in a rabbit model. RESULTS: In the present study, nano-titania-coated implants showed an increased coverage area and feature density, forming a homogenous layer compared to nano-HA implants. Bone contact values of the nano-titania implants showed a tendency to have a higher percentage as compared to the nano-HA implants (p = .1). CONCLUSION: Thus, no evidence of enhanced bone formation to nano-HA-modified implants was observed compared to nano-titania-modified implants. The presence of specific nanostructures dependent on the surface modification exhibiting different size and distribution did modulate in vivo bone response.

Nano hydroxyapatite structures influence early bone formation.

In a study model that aims to evaluate the effect of nanotopography on bone formation, micrometer structures known to alter bone formation, should be removed. Electropolished titanium implants were prepared to obtain a surface topography in the absence of micro structures, thereafter the implants were divided in two groups. The test group was modified with nanosize hydroxyapatite particles; the other group was left uncoated and served as control for the experiment. Topographical evaluation demonstrated increased nanoroughness parameters for the nano-HA implant and higher surface porosity compared to the control implant. The detected features had increased size and diameter equivalent to the nano-HA crystals present in the solution and the relative frequency of the feature size and diameter was very similar. Furthermore, feature density per microm(2) showed a decrease of 13.5% on the nano-HA implant. Chemical characterization revealed calcium and phosphorous ions on the modified implants, whereas the control implants consisted of pure titanium oxide. Histological evaluation demonstrated significantly increased bone formation to the coated (p < 0.05) compared to uncoated implants after 4 weeks of healing. These findings indicate for the first time that early bone formation is dependent on the nanosize hydroxyapatite features, but we are unaware if we see an isolated effect of the chemistry or of the nanotopography or a combination of both.

Formation of calcium phosphates on titanium implants with four different bioactive surface preparations. An in vitro study.

The aim of the present study was to compare the nucleating and growing behaviour on four types of bioactive surfaces by using the simulated body fluid (SBF) model. Titanium discs were blasted and then prepared by alkali and heat treatment, anodic oxidation, fluoridation, or hydroxyapatite coating. The discs were immersed in SBF for 1, 2, 4 and 6 weeks. Calcium phosphates were found on all specimens, as analysed with scanning electron microscopy/energy dispersive X-ray analysis (SEM/EDX). After 1 and 2 weeks of SBF immersion more titanium was accessible with SEM/EDX on the blasted surfaces than the four bioactive surface types, indicating a difference in coverage by calcium phosphates. The Ca/P mean ratio of the surfaces was approximately 1.5 after 1 week, in contrast to the fluoridated specimens which displayed a Ca/P mean ratio of approximately 2. Powder X-ray diffraction (P-XRD) analyses showed the presence of hydroxyapatite on all types of surfaces after 4 and 6 weeks of immersion. The samples immersed for 6 weeks showed a higher degree of crystallinity than the samples immersed for 4 weeks. In conclusion, differences appeared at the early SBF immersion times of 1 and 2 weeks between controls and bioactive surface types, as well as between different bioactive surface types.