Backscattered electron microscopy evidences tight apposition between bone and bioactive glass coating in vivo

Abstract The rationale for a degradable bioactive glass coating is to lead the bone to appose gradually to the metal. A bioactive glass already described in the literature (bg A) and a sodium-calcium-silicate non-bioactive glass sprayed as a control were studied. Young adult New Zealand White rabbits were selected as animal model. A hole was drilled from the femural intercondylar groove and Ti6Al4Vcoated cylinders were implanted. Retrieval took place at 1, 2, 4, 6, 8 and 10 months and samples were analyzed by backscattered electron microscopy (BSEM). For all the samples of bioactive glass, bone was in tight apposition with the coating. As time progressed, bone showed characters of physiological remodelling (newly formed bone substituting areas of bone resorption) close to the coating. At the interface between bone and bioactive glass coating, the apposition was so tight that it was not possible to discern a clear demarcation, even at higher BSEM magnification. A second key feature, in the behavior of the bioactive glass coatings invivo, was the gradual degradation and the eventual apposition of bone directly to Ti6Al4V.     

Mechanical failure of hydroxyapatite- and polysulfone-coated titanium rods in a weight-bearing canine model.

Three types of material that have shown potential as coatings for orthopaedic implants were studied. Using a weight-bearing canine model, Ti-6A1-4V femoral intramedullary rods coated with (1) sintered titanium beads, (2) plasma-sprayed hydroxyapatite, and (3) silyl coupled polysulfone beads were evaluated for mechanical strength and bone ingrowth. The model was designed to secure optimal prosthetic stability by obtaining maximal bony ingrowth during an initial non-weight-bearing phase, then stressing the implant during a full-weight-bearing phase. None of the rods coated with titanium beads failed. All 17 polysulfone-coated rods failed, 13 of them at the interface between the polysulfone coating and the titanium core. Of 18 rods coated with hydroxyapatite, 15 suffered implant breakdown at the interface between the hydroxyapatite coating and the titanium core. This may be due to dissolution of the plasma-sprayed hydroxyapatite in vivo. Testing of retrieved specimens from both hydroxyapatite- and polysulfone-coated implants showed that the shear strength at the coating-rod interface had decreased to less than 40% of the shear strength at manufacture. Despite mechanical failure, histologic study showed extensive bone ingrowth or apposition onto both the polysulfone and hydroxyapatite coatings.     

Osseointegration of hydroxyapatite porous-coated femoral implants in a canine model.

This study evaluated the quality and quantity of osseointegration of two thicknesses of hydroxyapatite coating on press-fit, porous-coated titanium implants in a canine hip model. In 12 dogs, titanium press-fit porous-coated prostheses were implanted. The stems had a 50-microm thickness hydroxyapatite coating, 100-microm thickness hydroxyapatite coating, or no hydroxyapatite coating. The animals were randomized into one of three groups and received one of the three implants. The implants were retrieved and examined 4 months after implantation. Direct juxtaposition of bone to the surface of the hydroxyapatite-coated implants with no intervening fibrous tissue layer was observed. There was no histologic evidence that hydroxyapatite initiated any foreign body reaction, nor was there any irregularity or resorption of the hydroxyapatite coating. There was a statistically significant greater degree of total bone apposition and bone ingrowth in the implants coated with hydroxyapatite at the level of the isthmus and the calcar. No statistical difference was found between the two groups with hydroxyapatite coatings in the degree of bone ingrowth or bone apposition.     

Hydroxyapatite-coated femoral stems. Histological analysis of components retrieved at autopsy.

Plasma-sprayed coating of hydroxyapatite are biocompatible and, because of their osteoconductive properties, may contribute to the early fixation of total joint prostheses. To evaluate this interface, we histologically analyzed five hydroxyapatite-coated femoral stems which, along with the surrounding bone, were retrieved from three humans at autopsy. The five femoral components had been in situ for a mean duration of twelve months (range, almost five to twenty-five months) and had been inserted for osteonecrosis (two), osteoarthrosis (two), and as an uncermented revision for failure of a cemented stem. The three patients had had a good or excellent clinical result and had died of causes unrelated to the joint arthroplasty. A coating of hydroxyapatite was identified on each stem. There was a variable amount of apposition of bone (32 to 78 per cent of available surface per section). The deposition of bone was most prominent on the surface of the prosthesis that was close to the endosteal surface of the bone, especially in areas that are predicted by Wolff's law (anterior and medial aspects of the implant, and at lateral-oblique corners). There were occasional foci of bone-remodeling around the implant, including osteoclast-mediated removal of the coating of hydroxyapatite along with adjacent bone. Occasional particles of ceramic were present within macrophages in the adjacent bone marrow. Other areas showed formation of new bone with a few areas of bone directly against the metal substrate. The over-all histological features suggest mechanically stable implants with bone-remodeling at the surface of the bone-implant interface.     

Alendronate does not inhibit early bone apposition to hydroxyapatite-coated total joint implants: a preliminary study

BACKGROUND: Alendronate is a pyrophosphate analogue of bisphosphonate that has been shown to inhibit osteoclastic bone resorption. Bone formation and remodeling are necessary to establish initial fixation of uncemented implants, especially those coated with a bioactive surface such as hydroxyapatite. Because the process of bone-remodeling that culminates in new-bone formation is thought to be initiated by osteoclastic bone resorption, it is appropriate to test the influence of osteoclast-inhibiting medications on bone apposition to hydroxyapatite-coated implants. METHODS: Twelve dogs underwent staged bilateral total hip arthroplasty, with twenty weeks between the first and second operations, with use of a titanium-alloy femoral stem that had a proximal macrotextured surface and a plasma-sprayed hydroxyapatite coating. Six of the dogs received oral alendronate therapy from the time of the surgery until they were killed; the other six dogs were untreated controls. The animals were killed four weeks after the second operation. Sections from matched implant sites (proximal, middle, and distal) were histologically analyzed. The linear extent of bone apposition, the linear extent and the thickness of the hydroxyapatite coating, and the total amount of cortical and trabecular bone were measured with the use of an interactive image analysis system. RESULTS: There were no significant differences in radiographic or histologic findings between the two groups at either four or twenty-four weeks. Although the extent of the hydroxyapatite coating decreased significantly with time in both groups (p < 0.01), we identified no significant influence of alendronate on the extent of bone apposition, the extent or thickness of the hydroxyapatite coating, or the cortical or trabecular bone area around the implants. CONCLUSIONS: Many patients who are receiving alendronate for osteoporosis or other disorders may also be candidates for cementless total joint arthroplasty. Although bone formation is generally thought to be initiated by and coupled with bone resorption, our results suggest that alendronate has no discernible effect on the initial fixation of or the short-term bone-remodeling around hydroxyapatite-coated femoral total joint implants.     

Bone ingrowth to implant surfaces in an inflammatory arthritis model

Many studies have shown enhanced bone apposition to implants coated with hydroxyapatite, but the optimum implant texture, especially in abnormal trabecular bone, is unclean The purpose of this project was to evaluate the histological and mechanical properties of cylindrical implants with three different surface textures that were placed in the cancellous bone of the distal femur of the rabbit after the production of an inflammatory knee arthritis. The three implant surfaces included a beaded surface (Group A), a beaded surface coated with hydroxyapatite (Group B), and a smooth surface coated with hydroxyapatite (Group C). The right knees of 36 rabbits were injected with carrageenan twice a week for 2 weeks. Then bilateral implantations were performed, with 12 rabbits in each group receiving identical implants in the right and left knees. The rabbits were killed 6 weeks after surgery. Mechanical (push-out test) and histomorphometric analyses were performed to determine the quality and quantity of bone ingrowth. In Group A, there was virtually no direct contact (a 20–60-μm clearance) between the bone and the beaded surfaces. Direct contact between the bone and the implant surfaces was seen in Groups B and C. The thickness and number of trabeculae were smaller on the arthritic side than on the control side for all groups but were not different between groups for either the control or the arthritic side. Mechanical testing showed that the shear strength of the interface was weaker on the arthritic side in all groups. The results suggest that inflammatory arthritis induced by carrageenan may influence the quality of local bone (osteopenic changes) and hence compromise the bone apposition and mechanical stability of the interface between the implant and bone.     

Comparative study of human cancellous bone remodeling to titanium and hydroxyapatite-coated implants

The human cancellous bone response was compared in weight-bearing porous hydroxyapatite (HA) and titanium-coated implants placed in the distal medial femoral condyles of consenting staged bilateral knee patients. The Institutional Review Board approved study quantified the amount of bone ingrowth, the mineral apposition rate, and the bone mineral content. Results showed that the osteoconductive HA coating increased the amount of bone ingrowth by 8% (P=.018). The HA coating did not effect the mineral apposition rate of the bone but had an 8% lower bone mineral content at the implant interface (P=.042). The influence of HA coatings on human cancellous bone appears highly focal along the coating surface. Gaps of 50–500 μm filled with fibrous connective tissue were observed along the porous-coated surfaces of both implant types suggesting that HA coatings still require precision placement adjacent to human cancellous bone.     

Bone ingrowth into polymer coated porous synthetic coralline hydroxyapatite

Porous hydroxyapatite, converted hydrothermally from the calcium carbon exoskeleton of the coral genus Goniopora (CHAG), has been shown to be effective as a scaffold for bone ingrowth (2,3,5-7,9). However, the large pores in the material resulted in low compressive strengths. In a previous study, we found that microcoating the internal surfaces of CHAG with dilactic-polyactic acid (DL-PLA) improved significantly its compressive properties. The objective of this study was to determine the effect of PLA microcoating on bone ingrowth into CHAG plugs. Plugs of thick- (3:1 chloroform to DL-PLA by weight), medium- (10:1), and thin- (30:1) coated as well as uncoated CHAG were implanted transcortically in the proximal third of the diaphysis of the rabbit tibia. Specimens were harvested at 3, 12, and 24 weeks for mechanical testing and contralaterally for histological and histomorphometric assessment. At 12 weeks, uncoated CHAG plugs developed an average ultimate interface shear stress of 26.7 MPa, compared with 17 MPa for 30:1 and 8 MPa for 10:1 and 3:1 coated specimens. At 24 weeks, there were no significant differences in shear stress among any of the specimens. Histomorphometric assessments showed that the ratio of area fractions of new bone to area fractions of new bone and void space increased from 68-70% for 3:1 and 10:1 coated specimens at 3 weeks, and to 85.5-89.5% at 24 weeks. In comparison, uncoated and 30:1 specimens had area fraction ratios of about 82% at 3 weeks and 93% at 24 weeks. Histologic sections demonstrated direct apposition of new bone to both the coating and the hydroxyapatite as well as degradation of the coating.     

Coating titanium implants with bioglass and with hydroxyapatite

This study compares the osteointegration of titanium implants coated with bioglass (Biovetro GSB formula) and with hydroxyapatite (HAP). Twenty-four bioglass-coated and 24 HAP-coated cylinders were implanted in the femoral diaphyses of sheep, and examined after 2, 4, 6, 8, 12, and 16 weeks. The HAP coating gave a stronger and earlier fixation to the bone than did bioglass. Bioglass formed a tissue interface which showed a macrophage reaction with little new bone formation activity. In contrast, HPA, showed intense new bone formation, with highly mineralised osseous trabeculae in the neighbourhood of the interface.     

Bone graft incorporation around titanium-alloy- and hydroxyapatite-coated implants in dogs.

To evaluate cancellous allogenic bone graft incorporation into porous-coated implants, the fixation of titanium alloy-(Ti) and hydroxyapatite-(HA) coated implants with and without bone graft was compared. An unloaded model with unilateral carragheenin-induced osteopenia of the knee was used in 12 mature dogs. Ti- and HA-coated cylinders were implanted in the distal femoral condyles and centralized in 2-mm overreamed drill holes. Allogenic, fresh-frozen (-80 degrees) cancellous bone graft was packed around the implants in six dogs. In a matched group of six other dogs, the implants were left in overreamed canals without bone graft. After six weeks the interface shear strength of grafted Ti-coated implants had significantly increased compared to the nongrafted Ti implants. However, HA coating used without bone graft was capable of enhancing the bone-implant interface shear strength to nearly the same degree. The fixation of grafted Ti- and HA-coated implants was equal. No significant difference in implant fixation was found between osteopenic and control bone. Histomorphometric evaluation of mineralizing surfaces in direct contact with the implant confirmed the results from the push-out test. Bone-implant fixation when using allogeneic fresh-frozen cancellous bone graft in osteopenic and control bone was enhanced by hydroxyapatite coating but the HA coating alone appeared to offer almost the same improvement in anchorage in 2-mm defects. Loss of bone stock around loose prosthetic implants often requires bone grafting. However, because of anatomic constraints in joint prosthetic surgery, a complete filling of defects with bone graft is difficult, and areas of gaps between bone and implant will remain. Provided mechanical stability of the prosthesis, the results reported here suggest that these areas will probably be filled early with new mineralizing bone if the prosthesis is coated with a thin layer of hydroxyapatite.     

Bonding of bone to apatite-coated implants

Implants of solid sintered hydroxyapatite form very tight bonds with living bone, but are susceptible to fatigue failure. This problem can be overcome by using plasma-sprayed apatite coatings on titanium implants. A very strong bond is formed between bone and this composite material; this was studied in canine bone with plug implants, avoiding any mechanical retention. Mechanical testing showed an interface shear strength at six weeks of 49 MPa with a maximum of 64 MPa after six months. There was histological evidence of direct bonding between the apatite coating and living bone while uncoated control plugs were easily extracted. The results indicate that apatite-coated implants can form a chemical fixation with a strength comparable to that of cortical bone itself. This fixation is far stronger than that provided by current cemented or uncemented fixation techniques.     

羟基磷灰石涂层钢板螺钉改善骨折稳定性的实验研究

目的　比较国产羟基磷灰石 (hydroxyapatite ,HA)涂层钢板螺钉与非涂层钢板螺钉在体负重情况下对骨折的内固定效果。方法　四孔钢板螺钉 2 4套 ,附加螺钉 60枚 ,对其中 12套钢板、30枚螺钉行HA等离子喷涂 ,其余的钢板、螺钉不喷。 12只成年杂种犬 ,每只犬的两侧肱骨线型骨折后分别以HA涂层钢板、螺钉或非涂层钢板、螺钉内固定 ,每只犬的右胫骨间隔 2cm交递置入HA涂层钉和非涂层钉共 5枚。按术后即刻、3、6周将犬分成 3组。处死后行组织学观察和生物力学测试。结果　组织学观察 :术后 3、6周组HA涂层均无明显降解 ,其与不锈钢基底结合良好 ,与骨组织结合紧密 ,3、6周组HA涂层钢板下都有不同程度的皮质骨上新生骨形成 ,该处未见明显的骨质疏松。相反非涂层钢板、螺钉与骨接触不紧密 ,其间有薄层纤维组织生长 ,钢板下可见不同程度的骨质疏松。力学测试 :3、6周组犬中HA涂层组的三点折弯强度及螺钉拔出强度均比非涂层组大。结论　国产HA涂层的钢板螺钉可增加骨折内固定的稳定性。     

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.     

Hydroxyapatite-coated porous titanium for use as an orthopedic biologic attachment system

The biologic attachment characteristics of hydroxyapatite (HA)-coated porous titanium and uncoated porous titanium implants were investigated. The implants were placed transcortically in the femora of adult mongrel dogs and evaluated after periods of three, six, and 12 weeks. The HA coating was applied using a modified plasma spray process to samples with pore volume and pore size of the porous coating expanded to equal the pore morphology of uncoated porous specimens. Mechanical push-out testing revealed that the bone-porous material interface shear strength increased with time in situ for both the uncoated and HA-coated implants. The use of the HA coating on porous titanium, however, did not significantly increase attachment strength. Histologic and microradiographic sections yielded similar qualitative results in the amount of bone grown into each system. After three weeks, both systems displayed primarily woven bone occupying approximately 50% of the available porous structure. Six and 12 weeks postimplantation, each system displayed more extensive bone ingrowth, organization, and mineralization, with only limited areas of immature bone. Histologically, differences were noted at the ingrown bone-porous material interface between the two implant types. The HA coating supported mineralization directly onto its surface, and a thin osseous layer was found lining all HA-coated surfaces. An extremely thin fibrous layer was observed separating the uncoated titanium particle surface from ingrown bone. There was no extensive direct apposition or lining of the ingrown bone to the uncoated porous titanium particle surfaces.     

Tissue reactions adjacent to well-fixed hydroxyapatite-coated acetabular cups. Histopathology of ten specimens retrieved at reoperation after 0.3 to 5.8 years

Ten acetabular cups coated with hydroxyapatite (HA) had originally been inserted in five primary and five revision total hip replacements. The thickness of the HA was 155 +/- 35 microm. The cups, which were well-fixed, were retrieved, with their adherent tissue, at reoperation after 0.3 to 5.8 years because of infection (five hips), wear of polyethylene (three hips), and instability (two hips). Undecalcified sections showed a direct contact between bone and osteoid-like tissue which had formed directly onto the HA coating. The area within the threads and their mirror images, as well as the implant-tissue interfaces consisted of similar amounts of bone and soft tissue. Degradation of HA was seen in all hips. The mean thickness of the remaining HA coating was 97 microm (95% CI 94 to 101). The metal interface comprised 66% HA. The HA-tissue interface contained more bone than soft tissue (p = 0.001), whereas the metal-tissue interface included more soft tissue than bone (p = 0.019). Soft tissue at the implant interface and poor replacement of HA by bone may interfere with long-term fixation.     

Pull-out strength and bone tissue reaction of plasma-sprayed hydroxyapatite coatings with different thicknesses or substrates]

Synthetic hydroxyapatite has been reported to have good histocompatibility, while metal to have good mechanical properties in vivo. This author coated hydroxyapatite on SUS316L stainless steel and titanium rods, using plasma-spraying techniques. Then they were implanted into tubular bones of five dogs. The pull-out strength of hydroxyapatite-coated metals was found to be stronger than that of non-coated metals and increased with passage of the time. After four weeks postoperatively, the coating layer fractured between hydroxyapatite and the metal by the pull-out test. A numerous new bone in contact with hydroxyapatite was observed. Fifty microns is found to be an ideal thickness of the coating layer. The pull-out strength of hydroxyapatite-coated titanium was higher, comparing to that of hydroxyapatite-coated SUS316L stainless steel. These results suggested that perovskite and rutile existed at the interface between hydroxyapatite and titanium after plasma-spraying and made hydroxyapatite to bond tightly to the titanium. Synthetic perovskite showed no pathological reactions to canine bone tissue.     

The morphogenesis of bone in replicas of porous hydroxyapatite obtained from conversion of calcium carbonate exoskeletons of coral.

The morphogenesis of bone in a porous hydroxyapatite substratum was studied after intramuscular implantation in adult primates. Replicas of porous hydroxyapatite that had been obtained after hydrothermal conversion of the calcium carbonate exoskeleton of coral (genus Goniopora) were implanted intramuscularly in twenty-four adult male baboons (Papio ursinus). Serial sections from specimens that had been harvested at three, six, and nine months showed that initially the formation of fibrous connective tissue was characterized by a prominent vascular component and by condensations of collagen fibers assembled at the interface of the hydroxyapatite. The morphogenesis of bone was intimately associated with the differentiation of the connective-tissue condensations. Bone formed without an intervening endochondral phase. Although the amount of bone varied considerably, in several specimens extensive bone developed, filling large portions of the porous spaces and culminating in total penetration by bone within the implants. The mean volume fraction composition of the specimens was 20.8 +/- 1.0 per cent (mean and standard error) for bone, 17.3 +/- 1.7 per cent for connective-tissue condensation, 31.9 +/- 1.0 per cent for fibrovascular tissue, 6.4 +/- 0.6 per cent for bone marrow, and 34.6 +/- 0.5 per cent for the hydroxyapatite framework. The amount of bone and marrow increased at each time-period, and the hydroxyapatite framework was significantly reduced between six and nine months. This indicated a moderate biodegradation over time, which was possibly a result of incomplete conversion of carbonate to hydroxyapatite. Linear regression analysis showed a negative correlation between the hydroxyapatite framework and the magnitude of bone formation within the porosities of the hydroxyapatite (p = 0.0001). Biochemical coating of the hydroxyapatite substratum with an allogeneic fibrin-fibronectin protein concentrate prepared from baboon plasma did not significantly increase the amount of bone formation within the porous spaces. The hydroxyapatite substratum may have functioned as a solid-phase domain for anchorage of bone morphogenetic proteins.     

Bonding of hydroxyapatite-coated femoral prostheses. Histopathology of specimens from four cases.

We examined specimens of hydroxyapatite-coated femoral prostheses from four patients who had died within nine months of implantation for fractured neck of femur. Histology showed newly formed immature bone overlying the hydroxyapatite coating with new trabeculae bridging to the endosteal bone layer. In the diaphysis, where there had been contact between the hydroxyapatite and the cortex, there was dense, firmly anchored bone with an haversian architecture. In other places the newly formed bone had a trabecular structure, containing bone marrow tissue with normal cellularity. It appeared that biological osseointegration had taken place.     

Hydroxyapatite coating enhances fixation of porous coated implants: A comparison in dogs between press fit and noninterference fit

Intimate contact at the bone-porous surface interface is not always achievable in noncemented prosthetic implantation. We investigated the effect of hydroxyapatite (HA) coating on skeletal attachment in non-interference fit 4 weeks after implantation in 6 mature dogs. The push-out test of HA-coated implants surrounded by a 1-mm gap showed a twofold increased shear strength and fivefold increased shear stiffness compared with titanium alloy (Ti) coated implants. The fixation of Ti implants was reduced by two thirds when inserted in a gap as compared with press fit, whereas HA-coated implants in gap showed anchorage close to implants in press fit. Only minor differences were found between HA and Ti implants in press fit. Histomorphometric analysis showed a significant increase in bone in direct contact to HA-coated implant as compared with Ti implants inserted both in gap and press fit. The study indicates that tightness of surgical fit is an important factor for sufficient fixation of the implant. However, our results demonstrate that hydroxyapatite coating almost eliminates the negative influence of noninterference fit between bone and unloaded implant.