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Bone formation in transforming growth factor beta-I-loaded titanium fiber mesh implants.

The osteoconductive properties of porous titanium (Ti) fiber mesh with or without a calcium phosphate (Ca-P) coating and osteoinductive properties of noncoated Ti fiber mesh loaded with recombinant human Transforming Growth Factor beta-1 (rhTGF-beta1) were investigated in a rabbit non-critical size cranial defect model. Nine Ca-P-coated and 18 non-coated porous titanium implants, half of them loaded with rhTGF-beta1, were bilaterally placed in the cranium of 18 New Zealand White rabbits. At 8 weeks postoperative, the rabbits were sacrificed and the skulls with the implants were retrieved. Histological analysis demonstrated that in the TGF-beta1-loaded implants, bone had been formed throughout the implant, up to its center, whereas in the non-loaded implants only partial ingrowth of bone was observed. Bone formation had a trabecular appearance together with bone marrow-like tissue. No difference in ingrowth could be observed between the non-TGF-beta1-loaded non-coated implants and the Ca-P-coated ones. All histological findings were confirmed by image analysis: 97% ingrowth was seen in the rhTGF-beta1-loaded implants, while only 57% and 54% ingrowth was observed in the non-loaded Ca-P-coated and non-coated implants, respectively. Bone surface area and bone fill were significantly higher in the rhTGF-beta1-loaded implants (1.37 mm2 and 36%, respectively) than in the non-loaded implants (0.57 mm2 and 26%). No statistical difference was found for any parameter between the Ca-P-coated and noncoated implants. Quadruple fluorochrome labeling showed that in the Ti and Ti-CaP implants mainly bone guidance had occurred from the former defect edge, while in the Ti-TGF-beta1 implants bone formation had mainly started in the center of a pore and proceeded in a centrifugal manner. Our results show that: (1) the combination of Timesh with TGF-beta1 can induce orthotopic bone formation; (2) Ti-fiber mesh has good osteoconductive properties; (3) a thin Ca-P coating, as applied in this study, does not seem to further enhance the bone-conducting properties of a titanium scaffold material.

Animals↗

Histomorphometric and microhardness assessments of sheep cortical bone surrounding titanium implants with different surface treatments.

Several factors influence the healing process and the long-term mechanical stability of cementless fixed implants, such as bone remodeling and mineralization processes. Histomorphometric and bone hardness measurements were taken in implants inserted in sheep femoral cortical bone at different times to compare the in vivo osseointegration of titanium screws (diam.; 3.5 x 7 mm length) with the following surface treatments: machined (Ti-MA); acid-etched (Ti-HF); HA vacuum plasma spray (Ti-HA); and Ca-P anodization followed by a hydrothermal treatment (Ti-AM/HA). Ti-MA and Ti-AM/HA implants presented the lowest (Ra = 0.20 +/- 0.01 microm) and highest (Ra = 1.97 +/- 0.64 microm) significant (p < 0.0005) roughness value, respectively. Bone-to-implant contact of Ti-HF was lower than that of the other surface treatments at both experimental times (8 weeks: -20%, ns; 12 weeks: -30%, p < 0.01). Significant differences in MAR (mineral apposition rate) were also found between the different experimental times for Ti-MA (115%, p < 0.01) and Ti-HF (57%, p < 0.01), demonstrating that bone growth had slowed inside the screw threads of Ti-HA and Ti-AM/HA after 12 weeks. No bone microhardness changes in preexisting host bone were found, while Ti-MA showed the lowest value for the inner thread area at 8 weeks (HV(200 microm)= 49.8 +/- 3.8 HV). These findings confirm that osseointegration may be accelerated by adequate surface roughness and bioactive ceramic coating such as Ca-P anodization followed by a hydrothermal treatment, which enhance bone interlocking and mineralization.

Animals↗

Static coefficient of friction between Ti-6Al-4V and PMMA for cemented hip and knee implants.

The static coefficient of friction between Ti-6Al-4V and PMMA was determined experimentally. A microtopographic surface analysis of the Ti-6Al-4V and PMMA specimens used in the experiments was performed to characterize the surfaces. The coefficient of friction between Ti-6Al-4V and PMMA in both dry and wet conditions, using both Ringer's solution and bovine serum, was determined by the standard inclined plane test, following the ASTM 4516-91 method, and by a prototype computerized sliding friction tester. The effects of surface roughness and of contact pressure on the coefficient of friction also have been investigated. Tests were performed at 26 degrees C and at body temperature of 37 degrees C. Considering all the tests, the overall range of the mean coefficients of friction varied between 0.17 and 0.32 in dry or wet conditions. For the same surface roughness in contact, in general the coefficient of friction using Ringer's solution was slightly lower than it was in dry conditions whereas bovine serum had a very high surface tension, which significantly increased the static coefficient of friction.

Alloys↗

Enhanced bone apposition around biofunctionalized sandblasted and acid-etched titanium implant surfaces. A histomorphometric study in miniature pigs.

Microrough titanium (Ti) surfaces of dental implants have demonstrated more rapid and greater bone apposition when compared with machined Ti surfaces. However, further enhancement of osteoblastic activity and bone apposition by bio-functionalizing the implant surface with a monomolecular adsorbed layer of a co-polymer - i.e., poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) and its derivatives (PLL-g-PEG/PEG-peptide) - has never been investigated. The aim of the present study was to examine early bone apposition to a modified sandblasted and acid-etched (SLA) surface coated with an Arg-Gly-Asp (RGD)-peptide-modified polymer (PLL-g-PEG/PEG-RGD) in the maxillae of miniature pigs, and to compare it with the standard SLA surface. Test and control implants had the same microrough topography (SLA), but differed in their surface chemistry (polymer coatings). The following surfaces were examined histomorphometrically: (i) control - SLA without coating; (ii) (PLL-g-PEG); (iii) (PLL-g-PEG/PEG-RDG) (RDG, Arg-Asp-Gly); and (iv) (PLL-g-PEG/PEG-RGD). At 2 weeks, RGD-coated implants demonstrated significantly higher percentages of bone-to-implant contact as compared with controls (61.68% vs. 43.62%; P < 0.001). It can be concluded that the (PLL-g-PEG/PEG-RGD) coatings may promote enhanced bone apposition during the early stages of bone regeneration.

Acid Etching, Dental↗

Healing of gaps around calcium phosphate-coated implants in trabecular bone of the goat.

Hydroxylapatite coatings are under clinical investigation in orthopaedics and dentistry. Bone formation on apatite coatings in the presence of gaps is important for clinical applications. The importance of the stability of the coating is not known at present. By varying the plasma-spray parameters, and by the addition of fluoride, the crystallinity and stability of calcium phosphates can be changed. It is suggested that bone formation is enhanced by dissolution of the apatite coating. We studied apatite coatings of varying stability with regard to their gap-healing characteristics, and we examined what the maximum gap would be that can be bridged if a coating is applied. Ti-6A1-4V implants coated with 62% crystalline hydroxylapatite, 30% crystalline hydroxylapatite or fluorapatite, or noncoated Ti-6A1-4V were implanted in 16 goats. The implants were surrounded by gaps of 1 or 2 mm, and the follow-up period was 6 weeks. Histological examination and histometry revealed that gaps of 1 mm can be bridged by bone if an apatite coating is applied. However, only a minimal amount of bone contact was seen on the apatite coatings with 2 mm gaps. Uncoated implants demonstrated no bone contact at all. Among the three different coatings there were no differences in gap healing. It can be concluded that in the goat, gaps of 2 or more mm between coated implants and host bone tissue inhibit bone deposition on the coating (p < 0.05), but the stability of the coating does not influence gap-healing characteristics.

Alloys↗

[Scanning electron microscopic study of titanium-hydroxyapatite, the functionally graded-material implanted in rabbits].

OBJECTIVE: The purpose of this study was to observe the tissue tolerance of Ti-HA functionally graded-material (FGM) and the form of the material-bone interface. METHODS: The sintered Ti -HA FGM, pure HA and pure Ti were respectively implanted into the parietal bone of rabbits. The specimens were observed by SEM at 2, 4, 8 postoperative weeks. RESULTS: In the early stage, the new bone surrounding the Ti -HA FGM formed earlier with larger amount and better maturity than the pure Ti. The condition was similar to the pure HA. Two months after the operation, direct bonding of material-bone interface was formed between the Ti -HA FGM and the new bone as an integral body. However, there was a little space left between the new bone and the pure Ti. CONCLUSIONS: The Ti -HA FGM has good tissue tolerance. Its early integration with bone is similar to pure HA and better than pure Ti.

Animals↗

Evaluation of the adaptation interface of one-piece implant-supported superstructures obtained in Ni-Cr-Ti and Pd-Ag alloys.

Several formulations of alternative alloys have been proposed for the substitution of gold-based alloys used in Dentistry. Recently, a Ni-Cr-Ti-based alloy has been introduced. The purpose of this work was to verify the marginal adaptation obtained with one-piece superstructures for implant-supported prostheses obtained in Ni-Cr-Ti alloy, compared to a semi-noble alloy Pd-Ag. Eight superstructures for each alloy were produced over 4 implants in the anterior region of the mandible. The superstructures were placed in a torquemeter and the fixation screw of implant #1 was tightened with a 20 Ncm load with the others loosened (Sheffield test). The unfitness (in mm) was measured using a three-dimensional optical measurer in each implant, in the buccal and lingual aspects. The obtained data were submitted to statistical analysis by the analysis of variance and Tukey's test at 5% level. Significant differences were found for the factors material (p< or =0.05), with Ni-Cr-Ti pieces better than Pd-Ag ones, and implants (p< or =0.01). There were no significant differences for the factor position and interactions among factors (p>0.05). Based on the analysis of the data, it is possible to conclude that the Ni-Cr-Ti alloy makes possible the obtainment of one-piece implant-supported superstructures with a smaller misfit compared to the one obtained in Pd-Ag alloy, traditionally indicated for this situation. Additional tests may verify the superiority of the Ni-Cr-Ti alloy.

Chromium Alloys↗

Response of normal female human osteoblasts (NHOst) to 17beta-estradiol is modulated by implant surface morphology.

Titanium (Ti) surfaces with rough microtopographies enhance osteogenic differentiation, local factor production, and response to osteogenic agents in vitro and increase pullout strength of dental implants in vivo. Estrogens regulate bone formation, resorption, and remodeling in females and may be important in implant success. Here, we tested the hypothesis that estrogen modulates osteoblast response to implant surface morphology. Primary female human osteoblasts were cultured to confluence on three Ti surfaces (pretreatment, PT - R(a) 0.60 microm; sandblasted and acid-etched, SLA - R(a) 3.97 microm; and Ti plasma-sprayed, TPS - R(a) 5.21 microm) and treated for 24 h with 10(-7) or 10(-8) M 17beta-estradiol (E(2)). Cell number decreased with increasing surface roughness, but was not sensitive to E(2). Alkaline phosphatase specific activity of isolated cells and cell layer lysates was lower on rough surfaces. E(2) increased both parameters on smooth surfaces, whereas on rough surfaces, the stimulatory effect of E(2) on alkaline phosphatase was evident only when measuring cell layer lysates. Osteocalcin levels were higher in the conditioned media of cells grown on rough surfaces; E(2) had no effect in cultures on the plastic surfaces, but increased osteocalcin production on all Ti surfaces. TGF-beta1 and PGE(2) production was increased on rough surfaces, and E(2) augmented this effect in a synergistic manner; on smooth surfaces, there was no change in production with E(2). The response of osteoblasts to surface topography was modulated by E(2). On smooth surfaces, E(2) affected only alkaline phosphatase, but on rough surfaces, E(2) increased levels of osteocalcin, TGF-beta1, and PGE(2). These results show that normal adult human female osteoblasts are sensitive to surface microtopography and that E(2) can alter this response.

Alkaline Phosphatase↗

Formation of highly adherent nano-porous alumina on Ti-based substrates: a novel bone implant coating.

Thin, nano-porous, highly adherent layers of anodised aluminium formed on the surface of titanium alloys are being developed as coatings for metallic surgical implants. The layers are formed by anodisation of a 1-5 microm thick layer of aluminium which has been deposited on substrate material by electron beam evaporation. The surface ceramic layer so produced is alumina with 6-8 wt % phosphate ions and contains approximately 5 x 10(8) cm(-2) pores with a approximately 160 nm average diameter, running perpendicular to the surface. Mechanical testing showed the coatings' shear and tensile strength to be at least 20 and 10 MPa, respectively. Initial cell/material studies show promising cellular response to the nano-porous alumina. A normal osteoblastic growth pattern with cell number increasing from day 1 to 21 was shown, with slightly higher proliferative activity on the nano-porous alumina compared to the Thermanox control. Scanning electron microscopy (SEM) examination of the cells on the porous alumina membrane showed normal osteoblast morphology. Flattened cells with filopodia attaching to the pores and good coverage were also observed. In addition, the pore structure produced in these ceramic coatings is expected to be suitable for loading with bioactive material to enhance further their biological properties.

Alloys↗

Ultrastructural features of the bone response to a plasma-sprayed hydroxyapatite coating in sheep.

The intentions of this study were to characterize the macroscopic, microscopic, and structural aspects of a plasma-sprayed implant and to thoroughly investigate bone tissue response after its implantation in sheep. Therefore, we used scanning electron microscopy, transmission electron microscopy (TEM), high-resolution TEM, X-ray diffraction, and energy-dispersive X-ray analyses. Assessment of the biomaterial prior to implantation showed a coating with irregular outlines and varying thickness, mainly consisting of hydroxyapatite (HA) covering a rough metallic implant core. Six months after insertion of the HA-coated Ti-6A1-4V implant, neither mechanical failure of the coating-substrate interface nor a significant loss of coating thickness was evident. However, an occasional lack of HA coating and phagocytosis of HA particles were noted. More generally, the implant was surrounded by well-mineralized bone investing the smallest cavities of the plasma-sprayed layer. Newly formed microcrystals with size, shape, and structure similar to those of bone apatite crystals were growing directly at the coating surface. These results suggest that the bone-bonding behavior of the considered grooved implant should provide satisfactory osseointegration and be suitable for fixed prostheses.

Animals↗

Enhancement of bone growth into metal screws implanted in the medullary canal of the femur in rats.

The kinetics of growth of bone into control (nontreated) and heat-treated screws made of stainless steel (type 316L) and Ti-6Al-4V, implanted in the medullary canal of the femur in rats, were studied by mechanical, histological, and biochemical methods. A progressive and significant increase in the ingrowth of bone, as reflected by interfacial shear strengths of the screws, was measured with time after implantation. At all time intervals for as long as 35 days after implantation, the shear strength of the heat-treated Ti-6Al-4V and stainless-steel screws was significantly higher than (1.6-3.4 times) that of the control screws. The specific activity of alkaline phosphatase in extracts of tissue from around the implanted screws peaked 6 days after insertion, with significantly higher values at 5, 6, and 7 days postoperatively for the heat-treated screws than for the controls. The extent of calcification also was higher at all time intervals for the heat-treated screws than for the controls. The histological evaluation of formation of bone between the ridges of the implanted screw corroborated the mechanical and biochemical measurements. At each time interval, a more mature bone was noted around the heat-treated screws than around the controls. It was concluded that the heat treatment of metal implants before insertion can result in augmentation of osseous ingrowth 1.6-5.3 times that into control implants in an in vivo experimental model.

Alkaline Phosphatase↗

The relevance of hydroxyapatite and spongious titanium coatings in fixation of cementless stems. An experimental comparative study in rat femur employing histological and microangiographic techniques.

Pure titanium rods plasma-spray coated with hydroxyapatite (HA) or porous titanium (Ti) of controlled roughness were implanted bilaterally in the distal femur of Sprague-Dawley rats to compare the extent of bone growth on the two types of coating. The relevance of other factors, like mechanical stability and biological adaptation of the bone to the insertion of a foreign body implant, were investigated in femora which were over-reamed (absence of primary fit) or reamed without insertion of the rod. Continuous tetracycline labeling for the first 30 days and for the last 2 weeks in the 90-day group was performed; histological/histometric, fluorescence and microangiographic studies were carried out on serial sections of the implanted and control femora. In the group of stable implants, HA-coated rods showed 90% integration versus 53% with Ti-coated implants (P < 0.001); in over-reamed implants neither surface bone growth nor endosteal fixation occurred, and both types of rods were surrounded by a thick layer of connective tissue. The study documented early adhesion of osteoblasts and direct deposition of bone matrix on the substrate, while on spongious titanium osteogenesis was observed only in proximity to the surface. Remodeling of the reactive, primary bone to mature, lamellar bone took the form of a capsule surrounding the implants and radial bridges connecting the latter to the endosteal surface. The number, height and thickness of these bridges appeared to be the factors determining implant stability, rather than the extent of the bony capsule on the perimeter of the implant. Integration was a function not only of mechanical conditions and surface geometry, but also of the biological response of the whole bone to changes in the vascularization pattern. The reported phenomena can be seen more easily in experimental models involving small rodents because of their fast bone turnover and revascularization, but it is expected that they take place, even at a lower speed, in clinical situations like cementless stems of total hip replacement.

Angiography↗

Quantitative and qualitative investigations of surface enlarged titanium and titanium alloy implants.

Screw shaped implants of commercially pure (c.p.) titanium and titanium-6aluminum-4vanadium (Ti6A14V) were blasted with particles of TiO2 of mean sizes of 25 microns (Group I) and 75 microns (Group II) and inserted in rabbit bone for 3 months. The surface roughness of the implants was examined and quantified with an optical scanning 3-dimensional instrument (TopScan 3D system), revealing the two alloy surfaces in each group had similar surface roughness. Biomechanical (removal torque) tests showed the c.p. titanium implants to be significantly more stable in the bone bed than those of Ti6A14V. In Group I, the c.p. titanium implants demonstrated a mean removal torque of 38 N cm while the Ti6A14V demonstrated a mean removal torque of 27 N cm (P = 0.004). Group II implants revealed a mean removal torque of 70 N cm for the c.p. ti and 50 N cm for the alloy samples (P = 0.003). The removal torque values were converted to shear forces/strengths by three calculation methods, based on (a) the entire length of the implant surface in the cortical region, (b) the thickness of the cortical bone measured in close vicinity to the thread peaks and (c) the bone-metal contact length measured on the non-unscrewed neighbouring implants. Group I: (a) the c.p. ti implants revealed a mean shear force of 4 vs a mean of 3 N/mm2 for the alloy samples. Shear strengths based on (b); were 8 for c.p. ti vs 6 N/mm2 for the alloy. The mean shear strength/force if calculated according to (c) revealed 23 for c.p. ti vs 18 N/mm2 for the alloy. Corresponding numbers for Group II; (a) c.p. ti 8 compared to 6 N/mm2 for the alloy, (b) c.p. ti demonstrated a mean value of 17 vs 11 N/mm2 for the alloy. According to method (c); c.p. ti had a mean shear strength of 26 vs 22 N/mm2 for the alloy samples. Histomorphometrical comparisons were performed on 10 microns thick undecalcified ground sections in the light microscope. In both Group I and Group II, the calculations of the mean bone-to-metal contact demonstrated more bone in contact to the c.p. titanium implants than to the Ti6A14V ones. Whereas comparisons of the bone volume inside the threads demonstrated slightly higher bone volumes around the alloy samples, no statistically significant difference was obtained between the two materials histomorphometrically.

Alloys↗

Tissue reactions to titanium endoprostheses. Autopsy studies in four cases.

Four cementless prosthetic hip implants of Ti-6Al-4V-alloy with and without longitudinal grooves and Al2O2 ceramic ball heads, with implant periods of 3, 4, 5, and 10 months, were sectioned and examined histologically in undecalcified thin ground sections. Especially on the medial and lateral sides of the prostheses, there was direct bone-to-metal contact without interposed connective tissue, spreading to the dorsal and ventral areas of the prosthesis shaft, which was considered osseointegration. This is based mainly on the technique of implantation with primary stability in the cortical bone by press-fit and on the bioinertness of the titanium alloy. Ungrooved prosthesis areas seem to favor bone growth at the shaft. The secondary bone ring, occurring on the dorsal and ventral areas of the prosthesis, with evidence of further bone remodeling with the passage of time, indicates that osseointegration is a dynamic process that continues to improve prosthetic stability.

Aged↗

In vitro and in vivo mechanical evaluations of plasma-sprayed hydroxyapatite coatings on titanium implants: the effect of coating characteristics.

This study was undertaken to evaluate the effect of coating characteristics on the mechanical strengths of the plasma-sprayed HA-coated Ti-6Al-4V implant system both in vitro and in vivo. Two types of HA coatings (HACs) with quite different microstructures, concentrations of impurity-phases, and indices-of-crystallinity were used. In vitro testings were done by measuring the bonding-strength at the Ti-6Al-4V-HAC interface, with HACs that had and had not been immersed in a pH-buffered, serum-added simulated body fluid (SBF). The shear-strength at the HAC-bone interface was investigated in a canine transcortical femoral model after 12 and 24 weeks of implantation. The results showed a bonding degradation of approximately 32% or higher of the original strength after 4 weeks of immersion in SBF, and this predominantly depended on the constructed microstructure of the HACs. After the push-out measurements, it was demonstrated that the HACs with higher bonding-strength in vitro would correspondingly result in significantly higher shear-strength at each implant period in vivo. Nevertheless, there were no substantial histological variations between the two types of HACs evaluated. The most important point elucidated in this study was that, among coating characteristics, the microstructure was the key factor in influencing the mechanical stability of the HACs both in vitro and in vivo. As a consequence, a denser HAC was needed to ensure mechanical stability at both interfaces.

Adhesiveness↗

[Research of bone bridge-like on plasma sprayed HA coating].

Bone formation on hydroxyapatite (HA) coating in the presence of gaps is important for clinical application. Pure Ti and hydroxyapatite coated by plasma sprayed samples were implanted in dog respectively. The implants were surrounded by gaps of 2 mm, and the follow-up period was 12 weeks. Histological examination and histomorphometry revealed that gaps could be bridged by bone provided the hydroxyapatite coating was applied, and that pure Ti implants were surrounded by fibrous tissue with no bone contact at all.

Animals↗

A light and scanning electron microscopic evaluation of electro-discharge-compacted porous titanium implants in rabbit tibia.

This study used light and scanning electron microscopic (SEM) histomorphometric methods to quantitate the rate of osseointegration of totally porous titanium alloy (Ti-6Al-4V) implants prepared by a novel fabrication technique--electrodischarge compaction (EDC). EDC was used to fuse 150-250-micrometer spherical titanium alloy beads into 4 X 6 mm cylindrical implants through application of a 300-microsecond pulse of high-voltage/high-current density. Two sterilized implants were surgically placed into each tibia of 20 New Zealand white rabbits and left in situ for periods corresponding to 2, 4, 8, 12, and 24 weeks. At each time point, 4 rabbits were humanely killed, and the implants with surrounding bone were removed, fixed, and sectioned for light and SEM studies. The degree of osseointegration was quantitated by means of a True Grid Digitizing Pad and Jandel Scan Version 3.9 software on an IBM PS/2 computer. The total pore area occupied by bone was divided by the total pore area available for bone ingrowth, and a Bone Ingrowth Factor (BIF) was calculated as a percent. The light microscopic results showed BIFs of 4% at week 2, 47% at week 4, 62% at week 8, 84% at week 12, and greater than 90% at week 24. The SEM results showed BIFs of 5% at week 2, 34% at week 4, 69% at week 8, 75% at week 12, and in excess of 90% at week 24. The results of this study show that EDC implants are biocompatible and support rapid osseointegration in the rabbit tibia and suggest that, after additional studies, they may be suitable for use as dental implants in humans.

Alloys↗