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Difference in penetration of horseradish peroxidase tracer as a foreign substance into the peri-implant or junctional epithelium of rat gingivae.

Horseradish peroxidase (HRP) tracer was applied to the gingival sulcus of implants or natural teeth at 5, 25, or 50 mg/ml to investigate the sealing capacities of the peri-implant epithelium (PIE) and junctional epithelium (JE); the extent of HRP penetration was observed under electron microscopy. A Ti-6Al-4V implant was inserted either immediately (immediate implantation) or 2 weeks (delayed implantation) after extraction of the maxillary left first molar of rats. The JE of the right molar was used as a control. Although the whole PIE of undecalcified sections appeared to be attached to the implant surface, electron microscopically, the internal basement lamina (IBL) and hemidesmosomes were deficient in the coronal-middle region of the PIE. There were extensive extracellular deposits of HRP in the intercellular spaces between PIE cells, and HRP was blocked to some extent by the lamina lucida and lamina densa of the external basal lamina and basal cell junction. HRP was detected in the connective tissue under the PIE, but was not found in the connective tissue under the JE. Intracellularly, HRP was found in the vesicles and granules of PIE cells and JE cells. These were fewer in number in PIE cells than in JE cells. There were no differences between the findings for immediate and delayed implantation. The results indicate that a deficiency in the IBL permitted penetration of HRP from the gingival sulcus into the connective tissue under the PIE, and suggest that the endocytotic capacity of PIE cells is inferior to that of JE cells.

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H2O2 production by cells on titanium and polystyrene surfaces using an in vivo model of exudate and surface related cell function.

The determination of secreted levels of reactive oxygen species by implant-adherent cells in vivo is required for understanding of the role(s) of such reactive oxygen species for the tissue response around medical devices. A model with subcutaneous implants of c.p. titanium (Ti) or polystyrene (PS) (cell culture grade) inserted on the back of rats were used. Implants and associated cells were retrieved and assayed after 1, 3, 5, 7, 14, 21 and 28 days. Morphological analysis of exudate cells showed that polymorphonuclear leukocytes (PMN) predominated after one day whereas macrophages were predominant after three days. The number of implant-adherent cells, as reflected by measurement of DNA, decreased with time. Ultrastructural observations showed that macrophages were predominant cells in contact with the implant surface. Measurement of hydrogen peroxide (H(2)O(2)) secretion by implant-adherent cells during 40 min incubation ex vivo revealed a constitutive generation of 40-400 pmol H(2)O(2)/microg DNA, depending on implantation time. Stimulation with protein kinase C agonist phorbol myristate acetate (PMA) caused an increased H(2)O(2) generation by adherent cells at early (up to five days) but not later (7-28 days) time periods. No major differences between Ti and PS were observed. Taken together, these findings show that Ti and PS implant-adherent cells secrete H(2)O(2) under in vivo conditions. Further, a reduced capacity to mount an enhanced H(2)O(2) secretion upon stimulation was demonstrated at late time periods. The role of this mediator for biocompatibility remains to be established.

Journal Article↗

Metal levels in cemented total hip arthroplasty. A comparison of well-fixed and loose implants.

In a prospective study, synovial fluid metal levels from stainless steel, cobalt-chromium, and titanium-alloy cemented total hip implants were measured. There were 37 well-fixed and 44 loose hip arthroplasties. Tissue-metal levels were quantitated in the cases revised for loosening. Retrieval analysis for implant wear was performed. Synovial fluid analysis showed a fivefold increase in metal levels of loose compared with well-fixed stainless steel implants. There was a sevenfold increase in metal levels of loose compared with well-fixed cobalt-chromium implants. There was a 21-fold increase in metal levels of loose compared with well-fixed titanium-alloy (Ti-6Al-4V) implants. Tissue-metal levels from revised cobalt-chromium implants averaged 45 micrograms/g dry tissue weight compared to 4,470 micrograms/g dry tissue weight from revised titanium-alloy implants, a 100-fold increase. Implant retrieval analysis showed severe burnishing and scratching in all titanium-alloy femoral heads and extensive burnishing and scratching in the majority of the femoral stems. Well-fixed cemented implants have similar low synovial fluid metal levels. However, when loosening of implants occurs, titanium-alloy implants release disproportionate levels of metal into synovial fluid and local tissues compared to stainless steel or cobalt-chromium.

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Fabrication and characterization of titanium-matrix composite with 20 vol% hydroxyapatite for use as heavy load-bearing hard tissue replacement.

Titanium-matrix composite with 20 vol% HA ceramic was fabricated by hot pressing technique and the microstructure of the composite was studied by transmission electron microscope (TEM). The mechanical and biological properties of the composite were investigated by mechanical and in vivo studies. The experimental results by TEM observation show the bonding state of Ti/HA interface in Ti-20 vol% HA composite with the relative density of 97.86% is good, however, there exists an interfacial transition zone between Ti and HA. In Ti matrix of the composite and pure Ti metal, an interesting substructure comprised of screw dislocations with Burgers vectors b of 1/3 < 11 20> was found. Screw dislocations are straight and regularly distributed, and cross slip can be observed. The subgrain boundaries consist of dislocation network walls with equidistant dislocation lines in the same direction. Elastic modulus and Vicker's hardness of Ti-20 vol%HA composite are 102.6 GPa and 3.41 GPa respectively. Owing to the existence of 20 vol% HA ceramic, bending strength and fracture toughness of the composite decrease sharply to 170.1 MPa and 3.57 MPa.m(1/2) respectively, which are only about 17.5 and 12% of those of pure Ti metal. In vivo studies indicate Ti-20 vol% HA composite has good biocompatibility, and even better osteointegration ability than pure titanium, especially in the early stage after the implantation. In conclusion, Ti-20 vol% HA composite is suitable for heavy load-bearing hard tissue replacement from the point of view of both mechanical properties and biocompatibility.

Animals↗

A histological comparison in the dog of porous-coated vs. threaded dental implants.

The histological findings of an 18-month trial, in the dog of a partially porous-coated endosseous dental implant made of Ti-6Al-4V, with a truncated conical shape, are described and compared with those for a cylindrical, threaded, endosseous implant made of commercially pure Ti. Six beagle dogs each received two porous-coated implants on one side of the mandible and two threaded implants on the contralateral side. Each set of two implants supported a two-unit fixed bridge for an 18-month functional period. Methylmethacrylate sections of both the buccolingual and mesiodistal aspects of each implant were examined qualitatively and by computer-assisted morphometry. The morphometric measurements were used for determination of the length of implant surface in direct contact with bone on each aspect of each implant. The data were expressed both as an absolute length and as a fraction of the maximum length available for contact (contact length fraction or CLF). On the buccal and lingual aspects of the implants, both the absolute lengths and CLF were significantly smaller for the porous-coated design. For the mesial and distal aspects, the absolute lengths and CLF were less for the porous-coated design, but the differences were not significant. However, when the absolute contact length was related to the corresponding vertical bone height, significant differences were observed, the absolute contact length being greater for any given bone height for the porous-coated design. Taken together, the data suggest that shorter implants may be used with the porous-coated design.

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Evaluating sol-gel ceramic thin films for metal implant applications. I. Processing and structure of zirconia films on Ti-6AI-4V.

Thin ceramic films or coatings over metallic bone-interfacing implant surfaces have the potential to improve implant performance with respect to implant fixation, wear, or corrosion. In this study, zirconia (ZrO2) thin films formed on Ti-6AI-4V using a polymeric alkoxide-based solgel process were investigated. ZrO2 films of uniform thickness on the order of 100 nm were obtained by dip coating Ti-6AI-4V samples into a zirconium propoxide containing solution using a substrate withdrawal speed ranging from 2 to 8 cm/min and a sol of nominal viscosity approximately 6 cps. These films were essentially free of surface macrodefects but had random submicron "pinholes." X-ray diffraction studies suggested that the films were at least partially crystalline, with some "metastable" cubic and/or tetragonal phases after annealing for 1 h at 500 degrees C. The demonstrated reproducibility of this approach for producing good quality ZrO2 films on Ti-6AI-4V warrants further studies to optimize processing conditions for implant applications.

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Evaluation of metallic and polymeric biomaterial surface energy and surface roughness characteristics for directed cell adhesion.

Directed cell adhesion remains an important goal of implant and tissue engineering technology. In this study, surface energy and surface roughness were investigated to ascertain which of these properties show more overall influence on biomaterial-cell adhesion and colonization. Jet impingement was used to quantify cellular adhesion strength. Cellular proliferation and extracellular matrix secretion were used to characterize colonization of 3T3MC fibroblasts on: HS25 (a cobalt based implant alloy, ASTM F75), 316L stainless steel, Ti-6Al4V (a titanium implant alloy), commercially pure tantalum (Ta), polytetrafluoroethylene (PTFE), silicone rubber (SR), and high-density polyethylene (HDPE). The metals exhibited a nearly five-fold greater adhesion strength than the polymeric materials tested. Generally, surface energy was proportional to cellular adhesion strength. Only polymeric materials demonstrated significant increased adhesion strength associated with increased surface roughness. Cellular adhesion on metals demonstrated a linear correlation with surface energy. Less than half as much cellular proliferation was detected on polymeric materials compared to the metals. However the polymers tested demonstrated greater than twice the amount of secreted extracellular matrix (ECM) proteins on a per cell basis than the metallic materials. Thus, surface energy may be a more important determinant of cell adhesion and proliferation, and may be more useful than surface roughness for directing cell adhesion and cell colonization onto engineered tissue scaffoldings.

3T3 Cells↗

A finite element model of the L4-L5 spinal motion segment: biomechanical compatibility of an interspinous device.

The biomechanical compatibility of an interspinous device, used for the "dynamic stabilization" of a diseased spinal motion segment, was investigated. The behaviour of an implant made of titanium based alloy (Ti6Al4V) and that of an implant made of a super-elastic alloy (Ni-Ti) have been compared. The assessment of the biomechanical compatibility was achieved by means of the finite element method, in which suitable constitutive laws have been adopted for the annulus fibrosus and for the metal alloys. The model was aimed at simulating the healthy, the nucleotomized and the treated L4-L5 lumbar segment, subjected to compressive force and flexion-extension as well as lateral flexion moments. The computational model has shown that both the implants were able to achieve their main design purpose, which is to diminish the forces acting on the apophyseal joints. Nevertheless, the Ni-Ti implant has shown a more physiological flexural stiffness with respect to the Ti6Al4V implant, which exhibited an excessive stiffness and permanent strains (plastic strains), even under physiological loads. The computational models presented in this paper seems to be a promising tool able to predict the effectiveness of a biomedical device and to select the materials to be used for the implant manufacturing, within an engineering approach to the clinical problem of the spinal diseases.

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The effect of calcium phosphate implant coating on osteoconduction.

OBJECTIVE: The purpose of this study was to determine whether a calcium phosphate (CaP) coating would have a significant impact on osteoconduction. STUDY DESIGN: In this investigation, porous-surfaced titanium alloy (Ti-6Al-4V) implants were prepared with or without the addition of a thin surface layer of CaP applied by means of sol-gel coating and implanted into the tibiae of 16 rabbits. Implant sites were allowed to heal for 2 weeks, after which specimens were retrieved for morphometric assessment by using backscatter scanning electron microscopy and the Bioquant Image Analyzer. RESULTS: The absolute contact length was significantly (P <.01) higher for CaP-coated implants (1.18 mm) than for the noncoated implants (0.74 mm), as were the contact length fraction (40.4% vs 27.0%; P <.01) and the straight-line bone growth (1.19 mm vs 1.04 mm; P <.01). CONCLUSION: On the basis of the findings in this study, the addition of a thin layer of CaP to the implant promotes accelerated bone healing around porous-surfaced implants-even after only 2 weeks of initial healing.

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Intramedullary implant of plasma-sprayed hydroxyapatite coating: an interface study.

An intramedullary implant model in the canine femora was developed to evaluate the mechanical and histological responses between cancellous bone and plasma-sprayed hydroxyapatite coatings (HACs) on ti-6A1-4V implants, with 12- and 24-week follow-ups. HACs of different thicknesses were investigated. Results of the mechanical testings revealed that after 24 weeks of implantation, the mean shear strength (2.49 +/- 0.12 MPa) of the 50 microns HACs was significantly higher (p < 0.05) than that of the 200 microns HACs (1.44 +/- 0.19 MPa). However, using backscattered electron images (BEIs) throughout all the implant periods, no substantial histological variations in the extent of new bone apposition between the two HACs were observed. Occasionally, solution-mediated disintegration of the 50 microns HAC was found 24 weeks postimplantation. Histomorphometric studies from the BEIs demonstrated that for both HACs the percentage of the direct HAC-cancellous bone contact was approximately 50% at 12 weeks and 75% at 24 weeks. After the mechanical tests, the 200 microns HACs had fracture sites either inside the coating layers or at the HAC-titanium interfaces, which might explain why the mechanical performance of the 200 microns HACs was inferior to that of the 50 microns HACs even though both HACs had the same histological behaviors.

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Tissue compatibility to different surfaces of dental implants: in vitro studies.

Factors affecting cell and tissue responses to dental implant biomaterials are typically characterized as spatial and temporal. Spatially, a dental implant must form an interface with both bone tissue for the development and maintenance of biomechanical stability and soft tissue for the prevention of microbial infection that can lead to peri-implantitis. There is a developing body of knowledge regarding the reactions of host tissues to implant materials, although the specific mechanisms by which these responses are not yet totally understood. From a temporal standpoint, there seems to be a sequence of events after placement of an implant that involves cell attachment, migration, and differentiation. These early wound healing responses appear to be influenced significantly by the properties of the underlying implant surface. Our laboratory has focused its attention on the first cellular event, i.e., cell attachment at the interface between bone and the implant surface. An in vitro primary cell model has been developed and used to study the influence of materials selection (Ti, Ti-6Al-4V, hydroxyapatite-like coatings), surface topography (smooth to rough), and surface chemistry (as a function of preparation treatments) on the cellular events that occur at implant surfaces. Currently, we believe that both uncoated Ti implants and those fabricated with calcium phosphate or hydroxyapatite-like coatings are capable of supporting initial cellular attachment, although they probably occur by different and incompletely understood mechanisms. The initial interactions of the host tissues with the implant surface remain key for long-term acceptance and must be understood if new generations of tissue engineered devices are to be developed and used clinically.

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Interface mechanics and histomorphometric analysis of hydroxyapatite-coated and porous glass-ceramic implants in canine bone.

A canine study was performed to make a histological and biomechanical evaluation of the interface between bone and two different bioceramic implants. A newly developed glass-ceramic formed by P2O5, CaO, SiO2, and Al2O3, giving a crystal phase composed of CaP2O6-AlPO4-SiP2O7, was compared to hydroxyapatite (HA) coated Ti-6Al-4V implants. A total of 24 implants were inserted into the femoral condyle of 15 adult female golden retriever dogs weighing 20-25 kg. There was a 12 week follow-up. Implants were examined by mechanical testing, histology, histomorphometry, microradiograpic methods, and EDAX analysis. The ultimate shear strength for the HA-coated implants was significantly higher than in the glass-ceramic group. When these values were related to the histomorphometric measurements, the difference could be explained by the tissue-to-implant contact. The glass-ceramic showed direct contact only with nonmineralized, osteoid bone. The HA-coated implants, however, were integrated into the bone. The study indicated that porous glass-ceramic containing AlPO4 causes local osteomalacia and might not be suitable for clinical purposes.

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Microstructure of interface regions and mechanical properties of Ti/Al 2O 3 and Ti-alloy/Al 2O 3 joints for dental implants.

Titanium and alumina are very well suited as constituents of dental metal/ceramic implants because of their excellent biocompatibility and their special chemical and mechanical properties which can be exploited to tailor composite implant structures. However, prior attempts to join pure titanium without any intermediate layer to alumina ceramic led to unsatisfactory results mainly due to thermal expansion mismatch between both materials. Therefore we used recently developed Ti alloys containing 30%wt Ta or 40%wt Nb for manufacturing dental implants. Moreover, we studied two alternative methods to join pure titanium with alumina using intermediate layers to reduce internal stresses within the joint caused by thermal expansion mismatch. We examined the interface region of these joints by metallographic, mechanical, analytical, and electron microscopy methods. Additionally, a comparison of the properties of the hitherto investigated types of joints with a view of their applicability in dental implants is given. Promising results were obtained for Ti/alumina joints with Nb interlayers. The studies are continuing.

Aluminum Oxide↗

In vitro evaluation of amorphous calcium phosphate and poorly crystallized hydroxyapatite coatings on titanium implants.

Studies of various apatite coatings on metal orthopaedic prostheses suggest that coating dissolution may promote enhanced bone bonding. Little is known concerning the effects of crystallinity and the underlying roughness on calcium phosphate (Ca/P) coating dissolution rate. To address these issues, the surface chemistry of amorphous Ca/P and poorly crystallized hydroxyapatite (HA) coatings on "smooth" and "rough" titanium (Ti) alloy (Ti-6A1-4V) implants was studied following immersion in Hank's physiologic solution at pH 7.2 and 5.2 for 0-, 4-, and 12-week periods. Changes in Calcium (Ca) ion concentrations in the solutions, coating chemistry, and surface morphology were studied by ion selective electrode, x-ray diffraction (XRD), and scanning electron microscopy (SEM) respectively. The amount of Ca dissolved from Ca/P-coated implants was strongly dependent on the chemistry of the coating and less dependent on pH or time of incubation. The effect of the underlying surface (smooth vs. rough) was not significant. The poorly crystallized HA coating underwent the most degradation, greatest crystallographic alteration, and greatest surface film formation. The amorphous coating was more stable in the saline environment, and may be more suitable in vivo if coating longevity is desired. These results suggest that this in vitro method is an effective way of determining differences in HA coating integrity.

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In vivo study of stainless steel and Ti-13Nb-13Zr bone plates in a sheep model.

A sheep study was performed to compare the in vivo performance of bone plates of 316L stainless steel and a new titanium alloy, titanium + 13% niobium + 13% zirconium (Ti-13Nb-13Zr), which had been subjected to a diffusion hardening treatment to produce a blue, wear resistant surface. Bone plates and screws of stainless steel and diffusion hardened Ti-13Nb-13Zr were implanted in adult sheep, in one group (with unosteotomized femurs) for 16 weeks, and in the other (with osteotomized femurs) for 8 weeks. At harvest, the diffusion hardened Ti-13Nb-13Zr devices had superior fixation strength, with greater screw torque out strength and fewer loose screws. In the osteotomized animals, the femurs with diffusion hardened Ti-13Nb-13Zr plates had higher torsional strength after removal of the implants; however, the difference was not statistically significant. In the unosteotomized animals, the torsional strength of the femurs was identical for both materials. There was a slightly reduced incidence of infection (bacterial adhesion) for the sheep with diffusion hardened Ti-13Nb-13Zr implants. In a parallel in vitro study, the magnetic resonance imaging compatibility of Ti-13Nb-13Zr was significantly superior to that of stainless steel. This indicates that diffusion hardened Ti-13Nb-13Zr may be an attractive alternative material for osteosynthesis.

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Improved bone anchorage of hydroxypatite coated implants compared with tricalcium-phosphate coated implants in trabecular bone in dogs.

Tricalcium phosphate (TCP) and hydroxyapatite (HA) ceramic coatings are bioactive coatings that have been shown to stimulate bone apposition onto ceramic-coated implants. TCP and HA ceramics have well-documented differences in physical properties, but both types of ceramics are used for stimulation of bone ongrowth to cementless endo-prosthetic components clinically. However, little is known about the difference in osteoconductive properties between these coatings when inserted into trabecular bone in a controlled experimental situation. Unloaded cylindrical gritblasted titanium (Ti-6A1-4V) implants (6 x 10 mm) coated with either hydroxyapatite (HA) or tricalcium phosphate (TCP) ceramic were inserted into the proximal humerus of 20 skeletally mature dogs. The implants were initially surrounded by a 2 mm gap. Each animal received one HA-coated implant and one TCP-coated implant. All dogs were sacrificed 6 weeks after surgery. Results were evaluated by histomorphometry and mechanical push-out test. Push-out tests demonstrated that HA-coated implants were 10-fold stronger fixated in comparison to TCP-coated implant. Bone ongrowth was significantly higher for HA-coated implants compared to TCP-coated implants. Bone volume in the gap showed a tendency to less bone volume around HA-coated implants compared to TCP-coated implants but this difference was insignificant. As expected almost all of the TCP coating were resorbed after 6 weeks and almost none of the HA coating. HA-coated implants with a grit-blasted surface provide a favorable early mechanical implant anchorage most likely due to superior ceramic stability compared to TCP-coated implants.

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Migration of polyethylene particles around stable implants in an animal model.

The aim of this study was to test the hypothesis that a tight seal between bone and implant will eliminate the avenue of particle migration around stable implants. Three types of implants were used in rabbits (polished press-fit Ti-6Al-4V or plasma-sprayed hydroxyapatite [HA]-coated Ti-6Al-4V) or doughy stage polymethyl methacrylate (PMMA). Implants were placed in the condylar notch. Each animal received an intra-articular injection of high density polyethylene (PE) particles (10(8) in 0.4 mL; mean size 4.7 microns) at 4 and 6 weeks postoperatively. Eight weeks postoperatively, peri-implant tissues were examined for PE particles and osteolysis. In all cases, intracellular PE particles were seen at the bone-implant interface and within marrow. No osteolysis was observed. Bone apposition was determined by computerized image analysis. There was no significant difference in the percentage of bone apposition (+/- SD) among the three groups of implants: Ti-6Al-4V (68% +/- 19%), HA-coated Ti-6Al-4V (70% +/- 10%), and PMMA (59% +/- 12%). These results indicate that a polished Ti-6Al-4V surface is as effective as PMMA or HA coating in limiting migration of PE particles around stable osseointegrated implants in rabbits.

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Histomorphometric comparison of implant anchorage for two types of dental implants after 3 and 6 months' healing in baboon jaws.

STATEMENT OF PROBLEM: A complete understanding of dental implant prognosis requires better knowledge of the bone anatomy after implant healing. Such baseline data are necessary to compare against load-induced changes in anatomy. PURPOSE: The purpose of this article is to describe and compare measures of implant support (percentage [%] integration and percentage [%] bone area) for various implants in baboon jaws after healing times of 3 and 6 months. MATERIAL AND METHODS: Commercially pure titanium (cpTi) and titanium alloy (Ti-alloy) screw-shaped implants were placed in the posterior jaws of 9 female baboons after 2 months of postextraction healing. Specimens were harvested after 3 months (5 baboons: 8 cpTi, 7 Ti-alloy) and after 6 months (4 baboons: 8 cpTi, 8 Ti-alloy). Each implant provided 6 polished horizontal sections for data collection, which was accomplished from digitized images with the IMAGE analysis system (reliability at 1.6%). Three- and six-month data for each parameter were compared with the use of ANOVA (P<.01). RESULTS: The results revealed a significant increase in the % integration (cpTi 39.1 to 56.2; Ti-alloy 40.0 to 55.2) and the % bone area (cpTi 38.8 to 47.9; Ti-alloy 38.9 to 49.2) from 3 to 6 months for both implants. This significant increase was also true for comparisons by jaw for each implant material (P<.01 for overall and by jaw comparisons). CONCLUSION: A time-dependent increase in jawbone anchorage was measured in this nonhuman primate population, and it was shown that the 6-month maxillary data were comparable to the 3-month mandibular data. These results lend support to the clinical strategy of waiting longer to load implants in the maxilla.

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