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.     

钙磷陶瓷涂层种植体—骨界面的成骨反应的定量实验研究

本文以恒河猴为实验对象,用未涂层纯钛种植体作为对照,用~(45)Ca示踪测定法及定量组织学方法研究两种生物玻璃陶瓷(MA_3)涂层种植体植入股骨后,种植体周围的骨盐沉积及新骨生长情况。结果表明:两种生物陶瓷涂层种植体的周围骨组织的钙盐沉积活跃期较未涂层者早,M-涂层种植体植入后1个月,其种植区的钙盐沉积程度就达到较高的水平。另外,在界面骨生长量方面,M-涂层植体的界面骨生长量在种植3个月内迅速增加,虽然A_4-涂层种植体的界面骨生长量在种植后3个月内高于未涂层者,但两者均通过3个月以后的继续增加,才接近于M-涂层种植体。     

Histologic evaluation of the osseous adaptation to titanium and hydroxyapatite-coated titanium implants.

The aim of this study was to obtain more information about the bone reaction to titanium and hydroxyapatite (HA)-coated titanium implants during the first 3 months after implantation. Therefore, uncoated and coated implants were inserted into the tibia of rabbits for various implantation periods. The histological results demonstrated that although there were no marked differences in bony reaction at the cortical level to the different implant materials, HA-coating appeared to induce more bone formation in the medullary cavity. It was also noted, that 3 months after insertion loss of coating thickness had occurred.     

Characterization of nano hydroxyapatite/collagen surfaces and cellular behaviors

Osseointegration at the bone-implant interface is a prerequisite for endosseous implants to succeed in achieving and maintaining their long-term stability in bone tissue. The achievement of osseointegration is significantly affected by surface nature of implants. To optimize osseointegration, this study presents the characterization of synthesized nanocrystalline hydroxyapatite (nano HA) and in vitro studies on nano HA, nano-HA/collagen, and titanium surfaces. Voids were found within the grain of nano HA, which consisted of the shell and the core. The finding assists the clarification of microstructures of nano HA. By low-temperature mixing nano-HA sol with collagen gel (nano-HA/collagen 80:20), nano HA, and nano-HA/collagen coated on pure titanium or porous anodic titanium oxides resulted in higher wettability and lower roughness. The in vitro studies showed that porous structures produced by anodic oxides on titanium served as positive anchorage sites for cell filopodia to connect, and nano HA decreased cell attachment of osteoblasts and induced well-developed long filopodia and broad lamellipodia, thereby enhancing cellular motility. Collagen involvement enhanced cell adhesion to nano HA. Cell reactions to nano HA, nano-HA/collagen, native, and porous titanium surfaces provide some guidance for an optimal osseointegration by their application in surface modifications for implants.     

Hydroxyapatite grafting promotes new bone formation and osseointegration of smooth titanium implants

Titanium is the ideal metal for intra-osseous dental implants. It permits the natural formation of an oxide layer on its surface and thereby it prevents the release of potentially toxic molecules. New formation of bone around implants, partially placed into the bone marrow cavity, is a gradual process that runs from the endosteum to the surface of the implant. Deposition of hydroxyapatite crystals on collagen type I fibrils is initiated by acidic proteins and leads to bone mineralization. This study analyzed the effects of hydroxyapatite upon peri-implant bone formation after insertion of smooth titanium implants. Screw-shaped smooth titanium implants of 3.75 mm thickness and 8.5 mm length were inserted into the metaphysis of rabbit tibia, either together with bovine hydroxyapatite into the right tibia or in controls without hydroxyapatite into the left tibia. Polyfluorochrome tracers (alizarin complex, calcein, tetracycline) were injected subcutaneously at different time intervals after implantation to evaluate the time frame of bone new formation over a period of 8 weeks. All samples were processed for histology and analyzed by fluorescence and polarizing microscopy. Our results showed a higher quantity of mature type I collagen fibers around implants and an acceleration of bone formation in the presence of hydroxyapatite. Mainly immature organic matrix was formed at the surface of implants in controls. The presence of hydroxyapatite seems to promote the maturation of collagen fibers surrounding the titanium implants and to support osteoconduction. Moreover, new formation of bone was faster in all samples where implants were inserted together with hydroxyapatite.     

Sphene ceramics for orthopedic coating applications: An in vitro and in vivo study

The host response to titanium alloy (Ti–6Al–4V) is not always favorable as a fibrous layer may form at the skeletal tissue–device interface, causing aseptic loosening. Recently, sphene (CaTiSiO₅) ceramics were developed by incorporating Ti in the Ca–Si system, and found to exhibit improved chemical stability. The aim of this study is to evaluate the in vitro response of human osteoblast-like cells, human osteoclasts and human microvascular endothelial cells to sphene ceramics and determine whether coating Ti–6Al–4V implants with sphene enhances anchorage to surrounding bone. The study showed that sphene ceramics support human osteoblast-like cell attachment with organized cytoskeleton structure and express increased mRNA levels of osteoblast-related genes. Sphene ceramics were able to induce the differentiation of monocytes to form functional osteoclasts with the characteristic features of f-actin and α_vβ₃ integrin, and express osteoclast-related genes. Human endothelial cells were also able to attach and express the endothelial cell markers ZO-1 and VE-Cadherin when cultured on sphene ceramics. Histological staining, enzyme histochemistry and immunolabelling were used for identification of mineralized bone and bone remodelling around the coated implants. Ti–6Al–4V implants coated with sphene showed new bone formation and filled the gap between the implants and existing bone in a manner comparable to that of the hydroxyapatite coatings used as control. The new bone was in direct contact with the implants, whereas fibrous tissue formed between the bone and implant with uncoated Ti–6Al–4V. The in vivo assessment of sphene-coated implants supports our in vitro observation and suggests that they have the ability to recruit osteogenic cells, and thus support bone formation around the implants and enhance osseointegration.     

Effect of hydroxyapatite/tricalcium-phosphate coating on osseointegration of plasma-sprayed titanium alloy implants

This study determined the effects of a plasma-sprayed hydroxyapatite/tricalcium phosphate (HA/TCP) coating on osseointegration of plasma-sprayed titanium alloy implants in a lapine, distal femoral intramedullary model. The effects of the HA/TCP coating were assessed at 1, 3, and 6 months after implant placement. The HA/TCP coating significantly increased new bone apposition onto the implant surfaces at all time points. The ceramic coating also stimulated intramedullary bone formation at the middle and distal levels of the implants. Fluorescent bone labeling indicated that new bone formation occurred primarily during the first 3 months after implantation, with comparatively little activity detected in the latter stages of the study. There was no associated increase in pullout strength at either 3 or 6 months; however, post-pullout evaluation of the implants indicated that the HA/TCP coating itself was not the primary site of construct failure. Rather, failure was most commonly observed through the periprosthetic osseous struts that bridged the medullary cavity. The demonstrated osteoconductive activity of HA/TCP coating on plasma-sprayed titanium alloy implant surfaces may have considerable clinical relevance to early host-implant interactions, by accelerating the establishment of a stable prosthesis-bone interface.     

In vitro investigation of titanium and hydroxyapatite dental implant surfaces using a rat bone marrow stromal cell culture system

In this study, rat bone marrow cells (RBM) were used to evaluate different titanium and hydroxyapatite dental implant surfaces. The implant surfaces investigated were: a titanium surface having a porous titanium plasma-sprayed coating (sample code Ti-TPS), a titanium surface with a deep profile structure (sample code Ti-DPS), an uncoated titanium substrate with a machined surface (sample code Ti-ma) and a machined titanium substrate with a porous hydroxyapatite plasma-sprayed coating (sample code Ti-HA). RBM cells were cultured on the disc-shaped test substrates for 14 days. The culture medium was changed daily and examined for calcium and phosphate concentrations. After 14 days specimens were examined by light microscopy, scanning electron microscopy, energy dispersive X-ray analysis and morphometry of the cell-covered substrate surface. All test substrates facilitated RBM growth of extracellular matrix formation. Ti-DPS and Ti-TPS to the highest degree, followed by Ti-ma and Ti-HA. Ti-DPS and Ti-TPS displayed the highest cell density and thus seem to be well suited for the endosseous portion of dental implants. RBM cells cultured on Ti-HA showed a delayed growth pattern. This may be related to its high phosphate ion release.     

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.     

The effect of surface macrotexture and hydroxylapatite coating on the mechanical strengths and histologic profiles of titanium implant materials

A mechanical and histological evaluation of uncoated and hydroxylapatite-coated titanium implant materials was performed. Cylindrical implants of uncoated commercially pure (CP) titanium and hydroxylapatite-coated Ti-6Al-4V alloy were studied using a transcortical model, with implants evaluated after periods of 3, 5, 10, and 32 weeks. All implants had a surface macrotexture consisting of a series of semicircular annular grooves, approximately 750 micron in maximum depth. The attachment characteristics of interface shear stiffness and interface shear strength were determined by mechanical push-out testing. Nondecalcified histologic and microradiographic techniques, with implants in situ, were used to evaluate the response to the implant materials and the presence of the surface macrotexture. Mechanical testing results indicated that the hydroxylapatite-coated implants exhibited significantly greater values of maximum interface shear strength than the uncoated implants after all time periods. Interface shear stiffness was also significantly greater at all time periods for the hydroxylapatite-coated implants as compared to the uncoated implants. Histological evaluation after 3 weeks revealed an osteoid layer covering on all areas coated with the hydroxylapatite material; mineralization of this layer appeared to be complete after 10 weeks. In all cases, longer-term implants demonstrated mineralization of interface bone directly onto the hydroxylapatite coating, and in no case was a fibrous layer observed between the hydroxylapatite coating and the interface bone. Sections from the uncoated CP titanium implants revealed a thin fibrous layer present in nearly all areas. Only isolated regions of direct bone-implant apposition were observed for the uncoated implants. The presence of this fibrous tissue layer, however, apparently did not adversely affect the development of considerable attachment strength. The results from this study indicate that the hydroxylapatite coating can significantly increase the attachment strength of implants which rely upon bone apposition for fixation. In addition, the hydroxylapatite coating provides an osteophilic surface for bone deposition, and allows for a more rapid development of implant-bone attachment.     

Stem cell-coated titanium implants for the partial joint resurfacing of the knee

The goal of the present study was to evaluate the partial surface replacement of the knee with stem cell-coated titanium implants and to provide a basis for a successful treatment of large osteochondral defects. Mesenchymal stem cells (MSCs) were isolated from bone marrow aspirates of adult sheep. Round titanium implants with a diameter of 2 x 7.3 mm were seeded with autologous MSC and inserted into an osteochondral defect in the medial femoral condyle. As controls, defects received either an uncoated implant or were left untreated. Nine animals with 18 defects were sacrificed after 6 months. Histological evaluation was performed by intravital polychrome fluorescent labelling, intravital perfusion with Indian ink, microradiographs and differential staining with toluidine blue. The quality of regenerated cartilage was assessed by in situ hybridization of collagen type II and immunohistochemistry of collagen types I and II. In 50% of the cases, defects treated with MSC-coated implants showed a complete regeneration of the subchondral bone layer. In these cases collagen type II and only traces of collagen type I were detected. A high level of collagen type II mRNA expression compared to articular cartilage indicates regenerating hyaline-like cartilage. A total of 50% of MSC-coated and uncoated implants failed to osseointegrate and formation of fibrocartilage was observed. Untreated defects as well as defects treated with uncoated implants demonstrated incomplete healing of subchondral bone and formation of fibrous cartilage. A modified histological score according to Wakitani significantly demonstrated better results for cell-coated implants (8.8+/-6.4) than for uncoated implants (5.5+/-3.9) and for untreated defects (2.8+/-2.5). Our results demonstrate that, in a significant number of cases, a partial joint resurfacing of the knee with stem cell-coated titanium implants occur. A slow bone and cartilage regeneration and an incomplete healing in half of the MSC-coated implants are limitations of the presented method. To improve our approach and optimize the experimental parameters, further investigations are needed prior to clinical application.     

The influence of BMP-2 and its mode of delivery on the osteoconductivity of implant surfaces during the early phase of osseointegration

Osteogenic agents, such as bone morphogenetic protein-2 (BMP-2), can stimulate the degradation as well as the formation of bone. Hence, they could impair the osteoconductivity of functionalized implant surfaces. We assessed the effects of BMP-2 and its mode of delivery on the osteoconductivity of dental implants with either a naked titanium surface or a calcium-phosphate-coated one. The naked titanium surface bore adsorbed BMP-2, whilst the coated one bore incorporated, adsorbed, or incorporated and adsorbed BMP-2. The implants were inserted into the maxillae of adult miniature pigs. The volume of bone deposited within a defined "osteoconductive" (peri-implant) space, and bone coverage of the implant surface delimiting this space, were estimated morphometrically 1-3 weeks later. After 3 weeks, the volume of bone deposited within the osteoconductive space was highest for coated and uncoated implants bearing no BMP-2, followed by coated implants bearing incorporated BMP-2; it was lowest for coated implants bearing only adsorbed BMP-2. Bone-interface coverage was highest for coated implants bearing no BMP-2, followed by coated implants bearing either incorporated, or incorporated and adsorbed BMP-2; it was lowest for uncoated implants bearing adsorbed BMP-2. Hence, the osteoconductivity of implant surfaces can be significantly modulated by BMP-2 and its mode of delivery.     

Bone growth enhancement in vivo on press-fit titanium alloy implants with acid etched microtexture

Early bone ongrowth secures long-term fixation of primary implants inserted without cement. Implant surfaces roughened with a texture on the micrometer scale are known to be osseoconductive. The aim of this study was to evaluate the bone formation at the surface of acid etched implants modified on the micro-scale. We compared implants with a nonparticulate texture made by chemical milling (hydrofluoric acid, nitric acid) (control) with implants that had a dual acid etched (hydrofluoric acid, hydrochloric acid) microtexture surface superimposed on the primary chemically milled texture. We used an experimental joint replacement model with cylindrical titanium implants (Ti-6Al-4V) inserted paired and press-fit in cancellous tibia metaphyseal bone of eight canines for 4 weeks and evaluated by histomorphometric quantification. A significant twofold median increase was seen for bone ongrowth on the acid etched surface [median, 36.1% (interquartile range, 24.3-44.6%)] compared to the control [18.4% (15.6-20.4%)]. The percentage of fibrous tissue at the implant surface and adjacent bone was significantly less for dual acid textured implants compared with control implants. These results show that secondary roughening of titanium alloy implant surface by dual acid etching increases bone formation at the implant bone interface.     

Biological performance of biomimetic calcium phosphate coating of titanium implants in the dog mandible

The aim of the present study was to analyze the in vivo effect of biomimetic calcium phosphate coating of titanium implants on periimplant bone formation and bone-/implant contact. Five types of implants were used: 1) Ti6Al4V implants with a polished surface; 2) Ti6Al4V implants with collagen coating; 3) Ti6Al4V implants with a mineralized collagen layer; 4) Ti6Al4V implants with sequential coating of hydroxyapatite (HA) and collagen; and 5) Ti6Al4V implants with HA coating only. All implants had square cross sections with an oblique diameter of 4.6 mm and were inserted press fit into trephine burr holes of 4.6 mm in the mandibles of ten beagle dogs. The implants of five animals each were evaluated after a healing period of 1 month and 3 months, respectively, during which time sequential fluorochrome labeling of bone formation had been performed. Bone formation was evaluated by morphometric measurement of the newly formed bone around the implants and the percentage of implant bone contact. After 1 month, there was a significantly higher percentage of mean bone/implant contact in the HA-coated implants compared to those with polished surface and those with the collagen-coated surface. After 3 months, these differences were not present anymore. Bone apposition was significantly higher next to implants with sequential HA/collagen coating compared to polished surfaces and mineralized collagen layer. It is concluded that biomimetic coating of titanium implants with HA has shown the clearest trend to increase bone-implant contact in the early ingrowth period. The addition of collagen to an HA coating layer may hold some promise when used as sequential HA/collagen coating with mineralized collagen as the surface layer.     

In vivo evaluation of anodic TiO2 nanotubes: an experimental study in the pig

Because of their ability to mimic the dimensions of constituent components of natural bone and the possibility to serve as a gene and drug-delivery carrier, nanotubes seem to be a promising coating for medical implants. Aim of this study was to investigate the effects of a TiO(2) nanotube structured surface on periimplant bone formation in vivo when compared with an untreated standard titanium surface. Twenty-five titanium implants covered with an ordered TiO(2) nanotube layer with an individual tube diameter of 30 nm and 25 commercially pure titanium (cp-Ti) implants were placed in the frontal skull of 25 domestic pigs. To evaluate the effects of the nanotube structured implants on the periimplant bone formation, bone-implant contact (BIC), and immunohistochemistry analysis were performed at day 3, 7, 14, 30, and 90. Evaluating immunohistochemistry, a significantly higher collagen type- I expression occurred at day 7 (p = 0.003), day 14 (p = 0.016), and day 30 (p = 0.044), for the nanostructured implants in comparison with the control group. It could be found that a nanotube structured implant surface with a diameter of 30 nm does influence bone formation and bone development by enhancing osteoblast function. SEM evaluation of the specimen surfaces revealed that the nanotube coatings do resist shearing forces that evoked by implant insertion. Because of their simple, low cost, flexible manufacturing and the possibility for the usage as drug or growth factor delivery system, nanotubes seem to be a promising method for future medical implant coatings.     

Permucosal implantation pilot study with HA-coated dental implant in dogs.

The histological response of transmucosal one-stage titanium dental implants coated with hydroxyapatite is described. The gingival adhesion to the implant was examined with regard to coated, partially coated or non-coated surfaces in the cervical region. From each coating type, 9 implants were inserted into dogs. Six months after the insertion, 19 implants could be evaluated, but 8 implants were lost. From these 19 implants, 6 implants showed severe pockets with inflammation up to the bony tissue. The 13 successful implants showed direct bone bonding with the hydroxyapatite coating and adhesion of submucosal connective tissue to the implant surface, with inflammation. The marginal gingiva showed slight inflammation. A totally coated implant will probably introduce inflammation by debris formation against the rough implant surface more easily. The hydroxyapatite coating often disappeared in the soft tissue or in the oral cavity. Bone which directly adapted to the coating seemed to prevent it from resorption.     

阳极氧化伴水热处理制备纯钛羟基磷灰石薄涂层

为了解纯钛阳极氧化伴水热处理的技术路线及羟基磷灰石薄涂层在体内的成骨效应，以钛片为阳极，β－磷酸甘油钠和醋酸钙为电解质，经200－400V直流电解15min，维持电流密度≤50mA/cm^2，而后经280℃－300℃水热处理2h，取光滑，喷砂表面作对照，分别植入12只兔股骨内，枯术后4，8，16周取出带种植体骨块制作磨片，观察新骨生成情况及术后8周光滑和涂层种植体的界面超微结构，并作表面能谱分析，结果发现水热处理后，钛片表面出现了羟基磷灰石涂层，动物实验显示涂层种植体8周后编织骨的转化和成熟较快，未见剥脱的羟基磷灰石碎片，其表面的钙，磷含量在种植后增加明显，这说明阳极氧化伴水热处理可以制备出纯钛表面羟基磷灰石薄涂层，其早期促编织骨形成作用明显，可以加快编织骨转化成板层骨。     

Histological and three-dimensional evaluation of osseointegration to nanostructured calcium phosphate-coated implants

Nanostructures on implant surfaces have been shown to enhance osseointegration; however, commonly used evaluation techniques are probably not sufficiently sensitive to fully determine the effects of this process. This study aimed to observe the osseointegration properties of nanostructured calcium phosphate (CaP)-coated implants, by using a combination of three-dimensional imaging and conventional histology. Titanium implants were coated with stable CaP nanoparticles using an immersion technique followed by heat treatment. Uncoated implants were used as the control. After topographical and chemical characterizations, implants were inserted into the rabbit femur. After 2 and 4weeks, the samples were retrieved for micro-computed tomography and histomorphometric evaluation. Scanning electron microscopy evaluation indicated that the implant surface was modified at the nanoscale by CaP to obtain surface textured with rod-shaped structures. Relative to the control, the bone-to-implant contact for the CaP-coated implant was significantly higher at 4weeks after the implant surgery. Further, corresponding 3-D images showed active bone formation surrounding the implant. 3-D quantification and 2-D histology demonstrated statistical correlation; moreover, 3-D quantification indicated a statistical decrease in bone density in the non-coated control implant group between 2 and 4weeks after the surgery. The application of 3-D evaluation further clarified the temporal characteristics and biological reaction of implants in bone.     

Nanocrystalline hydroxyapatite/titania coatings on titanium improves osteoblast adhesion

Bulk hydroxyapatite (HA) and titania have been used to improve the osseointegration of orthopedic implants. For this reason, composites of HA and titania have been receiving increased attention in orthopedics as novel coating materials. The objective of this in vitro study was to produce nanophase (i.e., materials with grain size less than 100 nm) HA/titania coatings on titanium. The adhesion of bone forming cells (osteoblasts) on the composite coatings were also assessed and compared with single-phase nanotitania and nano-HA titanium coatings. Nanocrystalline HA powders were synthesized through wet chemistry and hydrothermal treatments at 200 degrees C. Nanocrystalline titania powders obtained commercially were mixed with the nanocrystalline HA powders at various weight ratios. The mixed powders were then deposited on titanium utilizing a room-temperature coating process called IonTite. The results of the present study showed that such coatings maintained the chemistry and crystallite size of the original HA and titania powders. Moreover, osteoblasts adherent on single-phase nanotitania coatings were well-spread whereas they became more round and extended distinct filopodia on the composite and single-phase HA coatings. Interestingly, the number of osteoblasts adherent on the nanotitania/HA composite coatings at weight ratios of 2/1 and 1/2 were significantly greater compared with single-phase nanotitania coatings, currently-used plasma-sprayed HA coatings, and uncoated titanium. These findings suggest that nanotitania/HA coatings on titanium should be further studied for improved orthopedic applications.     

In vivo evaluation of bone-bonding of titanium metal chemically treated with a hydrogen peroxide solution containing tantalum chloride

Apatite formation on implants is important in achieving a direct bonding to bone tissue. We recently showed that titanium metal chemically treated with a hydrogen peroxide solution containing tantalum chloride has the ability to form a hydroxyapatite layer in simulated body fluid which had inorganic ion composition similar to human blood plasma. In this study, a pure titanium cylinder (4.0 mm in diameter, 20.0 mm in length) treated with this method was implanted into a hole (4.2 mm in diameter) in a rabbit's tibia. After implantation for predetermined periods up to 16 weeks, the specimens were extracted with bone tissue, and were examined by push-out test to evaluate the shearing force between the implant and bone tissue. The results were compared with those of non-treated pure titanium. Eight weeks after surgery, the shearing force of the treated titanium implanted in the 4.2 mm-hole was significantly higher than that of non-treated titanium, although the surface roughness was not changed after the treatment. Scanning electron microscopic (SEM) observation and energy-dispersive X-ray (EDX) microanalysis showed that the bone comes very close to the surface of the treated titanium. Moreover, the shearing force was higher for the implanted sample in the 4.0 mm-hole than that in the 4.2 mm-hole. Thus, it is confirmed that the treatment with hydrogen peroxide solution containing tantalum chloride provides higher bonding ability on titanium implants in vivo.