Bone ingrowth into two porous ceramics with different pore sizes: an experimental study

Many properties of porous calcium phosphate ceramics have been described, but how pore size influences bony integration of various porous ceramics remains unclear. This study was performed to quantify the bony ingrowth and biodegradability of two porous calcium phosphate ceramics with four different pore size ranges (45-80 microm, 80-140 microm, 140-200 microm, and 200-250 microm). Hydroxyapatite (HA) and beta-tricalcium phosphate (TCP) cylinders were implanted into the femoral condyles of rabbits and were left in situ for up to 12 months. The percentage of bone ingrowth and the depth of ingrowth within the pores were determined. Biodegradability of the implants was also evaluated. Bone ingrowth occurred at a higher rate into the TCP than into the HA ceramics with the same pore size ranges. The amount of newly formed bone was statistically smaller (p < 0.05) into ceramics with 45-80 microm pore size than with larger pore size, whatever the implantation time for HA and until four months for TCP. No statistical difference was noted between the three highest pore size ranges. No implant degradation was noted up to four months. Our results suggest that a pore size above 80 microm improves bony ingrowth in both HA and TCP ceramics. Bone formation was higher in the TCP than in the HA implants.     

Acid-etched microtexture for enhancement of bone growth into porous-coated implants

We designed an in vivo study to determine if the superimposition of a microtexture on the surface of sintered titanium beads affected the extent of bone ingrowth. Cylindrical titanium intramedullary implants were coated with titanium beads to form a porous finish using commercial sintering techniques. A control group of implants was left in the as-sintered condition. The test group was etched in a boiling acidic solution to create an irregular surface over the entire porous coating. Six experimental dogs underwent simultaneous bilateral femoral intramedullary implantation of a control implant and an acid etched implant. At 12 weeks, the implants were harvested in situ and the femora processed for undecalcified, histological examination. Eight transverse serial sections for each implant were analysed by backscattered electron microscopy and the extent of bone ingrowth was quantified by computer-aided image analysis. The extent of bone ingrowth into the control implants was 15.8% while the extent of bone ingrowth into the etched implants was 25.3%, a difference of 60% that was statistically significant. These results are consistent with other research that documents the positive effect of microtextured surfaces on bone formation at an implant surface. The acid etching process developed for this study represents a simple method for enhancing the potential of commonly available porous coatings for biological fixation.     

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.     

Mechanical and histological evaluation of hydroxyapatite-coated,titanium-coated and grit-blasted surfaces under weight-bearing conditions

Cylindric titanium rods with different surfaces were axially implanted into the femora of sheep. The three surfaces were grit-blasted titanium, plasma-sprayed titanium and plasma-sprayed hydroxyapatite (HA). After 2 months, a 2-cm segment of the femoral shaft was completely resected to load the implant, and the animals were allowed full weight-bearing for 9 months. Biomechanical and histological evaluation of the implants was undertaken 2 months after implantation and 9 months after the segmental resection. The mechanical testings of well-fixed implants were performed 9 months after segmental resection. Loosening of 45% of the titanium-coated implants was observed in the first 3 weeks, but thereafter, no further loosening occurred. The HA-coated implants remained entirely fixed for 3 weeks, but thereafter, a progressively increasing incidence of loosening up to 55% after 9 months of loading was detected as subsidence on X-radiographs. The maximum push-out strength of the titanium-coated implants was 4.9 MPa compared with 2.3 MPa for HA-coated ones. No significant mechanical interlock between the grit-blasted surface and bone was observed. The HA coating was found to be delaminated in all unstable implants, whereas the titanium coating remained completely intact. Morphometric analyses of well-fixed rods showed complete bony ingrowth onto the HA surface, whereas the contact area between the bone and the two titanium surfaces was less than 40%. Concerning clinical significance bony ingrowth with long-term mechanical interlock between the implant surface and the bone cannot be achieved by grit-blasting or HA-coating. The titanium plasma-coating, however, can induce a bone-implant interface which resists the mechanical stress resulting from continuous cyclic loading in vivo.Supported by Deutsche Forschungsgemeinschaft (Da-272/1-2)     

The bone-implant interface: a dynamic surface.

This study quantifies and compares bone formation on and around roughened titanium implants with roughened cobalt chromium, polished solid implants, and titanium fibermetal implants. Cylindrical rods were implanted into the medullary canal of the distal femur of rabbits. The bone-implant interface was studied 3, 6, and 12 weeks after surgery using histomorphometric methods. Roughened surface implants demonstrated significantly more bone directly apposed to the surfaces when compared to the polished or fiber/metal implants at 6 and 12 weeks after surgery. New bone formation and remodeling of bone occurred directly on roughened surfaces as late as 12 weeks after implantation, but not on the unroughened implants. These results suggest that roughening of the surfaces of both titanium and cobalt chromium implants can enhance osseointegration and may be useful clinically for the fixation of prosthetic components.     

Histomorphometric analysis of the repair of a segmental diaphyseal defect with ceramic and titanium fibermetal implants: Effects of bone marrow

We used a rat femoral diaphyseal defect/implant model to quantify the ingrowth of bone, cartilage, and fibrous connective tissue in a comparative study of woven sintered titanium fibermetal and porous hydroxyapatite/tricalcium phosphate ceramic implanted with and without the addition of syngeneic bone marrow cells. The patterns of tissue growth into the implants were analyzed with respect to time, type of implant, and the presence or absence of syngeneic marrow. Significantly more bone was found in ceramic implants than in fibermetal implants, with the addition of syngeneic marrow than without it, and at 4 months than at 2 months. Significantly more bone was found at both time periods in ceramic implants with bone marrow than in any other combination studied. We hypothesize that these findings resulted from interactions between the implanted material and its surroundings, specifically its ability to serve as a substratum for cell attachment, and cells in and around the defect, whether surgically implanted or arising from the soft-tissue bed.     

Locally delivered rhBMP-2 enhances bone ingrowth and gap healing in a canine model.

The purpose of the present study was to determine if recombinant human bone morphogenetic protein-2 (rhBMP-2) enhances bone ingrowth into porous-coated implants and gap healing around the implants. In the presence of a 3-mm gap between the implant and host bone, porous-coated implants were placed bilaterally for four weeks in the proximal humeri of skeletally mature, adult male dogs. In three treatment groups, the test implant was treated with HA/TCP and rhBMP-2 in buffer at a dose of 100 microg/implant (n=5), 400 microg/implant (n=6), or 800 microg/implant (n=5) and placed in the left humerus. In these same animals, an internal control implant was treated only with HA/TCP and buffer and placed in the right humerus. These groups were compared with a previously reported external control group of seven animals in which no growth factor was delivered [J. Orthop. Res. 19 (2001) 85]. The BMP treated implants in the two lower dose groups had significantly more bone ingrowth than the external controls with the greatest effect in the 100 g/implant group (a 3.5-fold increase over the external control, p=0.008). All three dose groups had significantly more bone formation in the 3-mm gap surrounding the BMP treated implants than the external controls with the greatest effect in the 800 microg group (2.9-fold increase, p<0.001). Thus, application of rhBMP-2 to a porous-coated implant stimulated local bone ingrowth and gap healing. The enhancement of bone formation within the implant (bone ingrowth) was inversely related to dose.     

Locally delivered rhTGF-beta2 enhances bone ingrowth and bone regeneration at local and remote sites of skeletal injury.

The purposes of the present study were to determine if recombinant human transforming growth factor-beta-2 (rhTGF-beta2) enhances bone ingrowth into porous-coated implants and bone regeneration in gaps between the implant and surrounding host bone. The implants were placed bilaterally for four weeks in the proximal humeri of skeletally mature, adult male dogs in the presence of a 3-mm gap. In three treatment groups of animals, the test implant was treated with hydroxyapatite/tricalcium phosphate (HA/TCP) and rhTGF-beta2 in buffer at a dose per implant of 1.2 microg (n = 6), 12 microg (n = 7), or 120 microg (n = 7) and placed in the left humerus. In these same animals, an internal control implant treated only with HA/TCP and buffer was placed in the right humerus. In a non-TGF-beta treated external control group of animals (n = 7), one implant was treated with HA/TCP while the contralateral implant was not treated with the ceramic. In vitro analyses showed that approximately 15%, of the applied dose was released within 120 h with most of the release occurring in the first 24 h. The TGF-beta treated implants had significantly more bone ingrowth than the controls with the greatest effect in the 12 microg/implant group (a 2.2-fold increase over the paired internal control (P = 0.004) and a 4-fold increase over the external control (P < 0.001)). The TGF-beta treated implants had significantly more bone formation in the gap than the controls with the greatest effect in the 12 and 120 microg groups (1.8-fold increases over the paired internal controls (P = 0.003 and P = 0.012, respectively) and 2.8-fold increases over the external controls (P < 0.001 and P = 0.001, respectively)). Compared to the external controls, the internal control implants tended to have more bone ingrowth (1.9-fold increase, P = 0.066) and had significantly more bone formation in the gap (1.7-fold increase. P = 0.008). Thus, application of rhTGF-beta2 to a porous-coated implant-stimulated local bone ingrowth and gap healing in a weakly dose-dependent manner and stimulated bone regeneration in the 3-mm gap surrounding the contralateral control implant, a site remote from the local treatment with the growth factor.     

Porous cups with and without hydroxylapatite-tricalcium phosphate coating: 23 matched pairs evaluated with radiostereometry

Migration, wear, and presence of radiolucencies were studied in 23 matched pairs of patients operated with porous-coated acetabular cups with additional screw fixation. All implants had the same type of titanium fiber mesh. In each pair, one of the cups was plasma-sprayed with a coating consisting of 70% hydroxylapatite (HA) and 30% tricalcium phosphate (TCP). Radiostereometric analysis up to 2 years after the operation revealed smaller rotations around the horizontal axis in cups with HA/TCP coating. The migration of the cup center was not significantly influenced. Evaluation of femoral head penetration in 12 of the matched pairs did not reveal any significant difference. Immediately after operation, implants with HA/TCP coating had more central radiolucencies, which, despite minimal migration, disappeared during the follow-up. The clinical results did not differ between the 2 groups. The findings of less tilting and diminishing radiolucencies in the cups with HA/TCP coating suggest a more complete ingrowth of bone and a better sealing of the interface.     

Comparison of hydroxyapatite and hydroxyapatite tricalcium-phosphate coatings

This study compared the effects of hydroxyapatite (HA) coating and biphasic HA/tricalcium-phosphate (HA/TCP) coating on the osseointegration of grit-blasted titanium-alloy implants. Each coated implant was compared with uncoated grit-blasted implants as well. The implants were press-fit into the medullary canal of rabbit femora, and their osseointegration was evaluated 3 to 24 weeks after surgery. The coated implants had significantly (P<.05) greater new bone ongrowth than the uncoated implants (HA, 56.1 +/- 3.1%; HA/TCP, 53.8 +/- 2.6%; uncoated, 32.2 +/- 1.4% of the implant perimeter, 12 weeks). Unmineralized tissue (cartilage and osteoid) was seen on the uncoated implants but never on the coated implants. The coated implants had significantly (P<.05) greater interfacial shear strength than the uncoated implants (HA, 4.1 +/- 0.4 MPa; HA/TCP, 4.8 +/- 0.5 MPa; uncoated, 2.6 +/- 0.2 MPa, 12 weeks). There was no difference between HA and HA/TCP coating in regard to new bone growth or interfacial shear strength. These data show a comparable enhancement effect of HA and HA/TCP coatings on the osseointegration of titanium-alloy implants.     

An in vivo evaluation of bone response to three implant surfaces using a rabbit intramedullary rod model

Our study was designed to evaluate osseointegration among implants with three surface treatments: plasma-sprayed titanium (P), plasma-sprayed titanium with hydroxyapatite (PHA), and chemical-textured titanium with hydroxyapatite (CHA). Average surface roughness (Ra) was 27 microns for the P group, 17 microns for the PHA group, and 26 microns for the CHA group. Bilateral distal intramedullary implants were placed in the femora of thirty rabbits. Histomorphometry of scanning electron microscopy images was used to analyze the amount of bone around the implants at 6 and 12 weeks after implantation. Greater amounts of osseointegration were observed in the hydroxyapatite-coated groups than in the noncoated group. For all implant surfaces, osseointegration was greater at the diaphyseal level compared to the metaphyseal level. No significant differences were seen in osseointegration between the 6 and 12 week time points. Although the average surface roughness of the P and the CHA groups was similar, osseointegration of the CHA implants was significantly greater. The results of this in vivo lapine study suggest that the presence of an hydroxyapatite coating enhances osseointegration despite similarities in average surface roughness.     

Enhanced bone ingrowth and fixation strength with hydroxyapatite-coated porous implants

The effect of a hydroxyapatite (HA) coating on the interface attachment strength, rate of development of attachment strength, and degree of bone ingrowth of porous implants was investigated. Implants with ideal surgical fits and those having interface gap spaces were evaluated using femoral transcortical and intramedullary models. The application of a thin HA coating to porous implants significantly enhanced both interface attachment strength and bone ingrowth. The rate of development of interface strength and bone ingrowth was also more rapid with the HA-coated system. There was no evidence of any disruption, loss, or resorption of the HA coating.     

Hydroxyapatite coating modifies implant membrane formation. Controlled micromotion studied in dogs.

We studied the influence of controlled micromovements between bone and porous titanium alloy implants with and without hydroxyapatite coating. A dynamically loaded unstable device producing approximately 150-microns axial translation of knee implants during each gait cycle was developed. Stable implants served as controls. Matched stable and unstable implants with either porous titanium (Ti) or hydroxyapatite (HA) coating surrounded by a gap of 0.75 mm were inserted into the weight-bearing regions of the medial femoral condyles in 14 mature dogs. Histologic analysis after 4 weeks showed a fibrous membrane surrounding both types of implants subjected to micromovements, whereas various amounts of bone ingrowth was obtained in the stable implants. The membrane around unstable HA implants was thinner than that around unstable Ti implants. Islands of fibrocartilaginous tissue characterized the membrane around unstable HA implants, whereas fibrous connective tissue surrounded unstable Ti implants. The collagen concentration of the fibrous membranes was higher around unstable HA implants compared with Ti implants. Instability reduced the shear strength of the implants. However, the shear strength of unstable HA implants exceeded that of the Ti implants, both unstable and stable. The greatest shear strength was obtained by stable HA implants, i.e., tenfold greater than that of stable Ti implants. The gap-healing capacity around stable HA implants increased toward the HA surface, and was greater than that around Ti implants. Our study demonstrates that micromovements between bone and implant inhibit bone ingrowth and lead to the development of a fibrous membrane. The superior fixation of unstable HA implants compared with unstable Ti implants may be ascribed to the presence of fibrocartilage, a higher collagen concentration, and radiating orientation of collagen fibers in the membrane. The strongest mechanical anchorage and the greatest amount of bone ingrowth was obtained by stable implants coated with hydroxyapatite.     

Bone ingrowth into porous calcium phosphate ceramics: influence of pulsing electromagnetic field

The effect of a pulsing electromagnetic field (PEMF) on bone ingrowth into porous hydroxyapatite (HA) and porous tricalcium phosphate (TCP) implanted in rabbit tibiae was studied. To quantitate the biological response, a recently developed method of surface measurement using a scanning electron microscope was used. The morphometrical findings in the HA pores demonstrated a significantly greater amount of bone and thicker bone trabeculae in the PEMF group as compared with the nonpulsed control group at 3 to 4 weeks postimplantation. No significant differences for these parameters were found in the TCP pores. Histologically, more bone and wider bone trabeculae were observed in the HA implants for the PEMF-treated animals at the early time periods when compared with those of the control animals. Alternatively, the histological findings of the TCP implants were similar between these two groups. These histological results tended to correlate with the morphometrical data. Together, these results suggest that accelerated bone formation and bone maturation occurred in response to PEMF in the HA pores but was without effect in the TCP pores. This stimulatory effect is most significant after 3-4 weeks of PEMF stimulation.     

Tissue ingrowth into titanium and hydroxyapatite-coated implants during stable and unstable mechanical conditions.

Lack of initial mechanical stability of cementless prostheses may be responsible for fibrous tissue fixation of prosthetic components to bone. To study the influence of micromovements on bony ingrowth into titanium alloy (Ti) and hydroxyapatite (HA)-coated implants, a loaded unstable device producing movements of 500 microns during each gait cycle was developed. Mechanically stable implants served as controls. The implants were inserted into the weight-bearing regions of all four femoral condyles in each of seven mature dogs. Histological analysis after 4 weeks of implantation showed a fibrous tissue membrane surrounding both Ti and HA-coated implants subjected to micromovements, whereas variable amounts of bony ingrowth were obtained in mechanically stable implants. The pushout test showed that the shear strength of unstable Ti and HA implants was significantly reduced as compared with the corresponding mechanically stable implants (p less than 0.01). However, shear strength values of unstable HA-coated implants were significantly greater than those of unstable Ti implants (p less than 0.01) and comparable to those of stable Ti implants. The greatest shear strength was obtained with stable HA-coated implants, which was threefold stronger as compared with the stable Ti implants (p less than 0.001). Quantitative determination of bony ingrowth agreed with the mechanical test except for the stronger anchorage of unstable HA implants as compared with unstable Ti implants, where no difference in bony ingrowth was found. Unstable HA-coated implants were surrounded by a fibrous membrane containing islands of fibrocartilage with higher collagen concentration, whereas fibrous connective tissue with lower collagen concentration was predominant around unstable Ti implants. In conclusion, micromovements between bone and implant inhibited bony ingrowth and led to the development of a fibrous membrane. The presence of fibrocartilage and a higher collagen concentration in the fibrous membrane may be responsible for the increased shear strength of unstable HA implants. Mechanically stable implants with HA coating had the strongest anchorage and the greatest amount of bony ingrowth.     

A long-term study on defect filling and bone ingrowth using a canine fiber metal total hip model.

When performing primary and revision total hip arthroplasty (THA), bone defects are often encountered. At present, grafting osseous defects with autogeneic bone is a common means of treatment. In this study, defects in bone were created in the femora and acetabula of dogs being treated with cementless THA with a fiber metal implant (Group A) or a hydroxyapatite tricalcium phosphate (HA/TCP) sprayed implant (Group B). The following methods of defect filling were compared: (1) leaving defects unfilled, (2) filling with autogeneic bone graft, (3) filling with a 50:50 mixture of autograft and a biphasic ceramic composed of HA/TCP, and (4) filling with a collagen-HA/TCP-bone marrow mixture. Analysis of defect healing and the extent of ingrowth into the overlying fiber metal, at defect sites and sites distant from defects, was made at six, 12, and 24 weeks postimplantation. Defect healing was enhanced at six and 12 weeks in all grafted groups when compared with ungrafted controls. Bone ingrowth into the porous fiber metal overlying the defects was not significantly affected by grafting the defects, compared with the ungrafted defects. The extent of bone ingrowth into the fiber metal acetabular implant at sites away from the defects increased during the entire study. In contrast, the extent of bone ingrowth on the femoral side was maximal at 12 weeks. The HA/TCP coating enhanced ingrowth into the acetabular component at 12 weeks, compared with the uncoated prosthesis, but did not enhance ingrowth on the femoral side. The data from this study demonstrate that defect healing is enhanced with graft materials. However, this does not necessarily result in increased ingrowth into porous surfaces overlying osseous defects. General bone ingrowth and ingrowth at defect sites at 12 weeks postimplantation can be enhanced on the acetabular side with the use of HA/TCP-sprayed implants. However, no positive effect is seen with the use of an HA/TCP-sprayed femoral implant.     

Porous hydroxyapatite and tricalcium phosphate cylinders with two different pore size ranges implanted in the cancellous bone of rabbits. A comparative histomorphometric and histologic study of bony ingrowth and implant substitution

To investigate the histophysiology of implant degradation, hydroxyapatite and tricalcium phosphate cylinders with a diameter of 3 mm were implanted in the cancellous bone of the distal femur and the proximal tibia of 15 New Zealand White rabbits for up to six months. All implants had a homogeneous pore distribution and a porosity of 60%. Ceramics with a pore size range of 50-100 micron and 200-400 micron were compared. Morphometric analysis showed that up to 85.4% of the originally implanted tricalcium phosphate was degraded after six months, whereas the volume reduction of the hydroxyapatite was only 5.4% after the same period. Within the first months bone and tissue ingrowth and implant resorption occurred at a higher rate in the smaller-pored tricalcium phosphate than in the larger-pored material. Hydroxyapatite cylinders with small pores were totally infiltrated by bone or bone marrow after four months, whereas in the larger-pored hydroxyapatite implants tissue did not penetrate all pores after six months and the amount of bone within the implant was small. Scanning electron microscopy of the material before implantation revealed the existence of numerous pore interconnections with diameters of about 20 micron in the smaller-pored ceramics. Such interconnections were rare in the larger-pored implants. The pore interconnections seem to promote vascular and tissue ingrowth and consequently the initial rate of implant resorption. Implant resorption is an active process and involves two different cell types. Acid phosphatase-positive osteoclast-like cells suggesting active resorption adhere directly to the surface, especially in tricalcium phosphate implants. Clusters of macrophages tightly packed with granular material are found in the pores and along the perimeter of all implant cylinders. They may play an active role in the intracellular degradation of small detached ceramic particles.     

Osseointegration in arthroplasty: can simvastatin promote bone response to implants?

Cementless fixation depends on bone ingrowth for long-term success. Simvastatin as a lipid lowering agent has been demonstrated to have osteoanabolic effects. This study was designed to measure the possible effect of simvastatin on implant osseointegration. Bilateral femoral implantation of titanium cylinders was performed in 20 rabbits. Blood lipid levels were measured pre- and postoperatively. Scanning electron microscopy (SEM) was used to measure the percentage of the surface of each implant in contact with bone and mechanical pull-out testing was performed. The blood lipid levels were significantly reduced in the simvastatin group. Histomorphometric examination revealed increased bone ingrowth and mechanical examination showed increased interface strength in the simvastatin group. Mechanical and histological data showed superior stability and osseous adaptation at the bone/implant interface for the simvastatin group. We conclude that simvastatin has potential as a means of enhancing bone ingrowth, which is a key factor in the longevity of cementless implants.     

Postmortem comparative analysis of titanium and hydroxyapatite porous-coated femoral implants retrieved from the same patient: A case study

The results of bilateral postmortem analysis of titanium and plasma-sprayed hydroxyapatite (HA) porous-coated femoral components of the same Anatomic Porous Replacement design retrieved from a 35-year-old female donor are reported. Analysis was conducted using backscattered electron imaging, histology, and radiographic techniques. The appositional bone index, percent bone ingrowth, and mineral content were measured for both implants. The results showed a 177% higher appositional bone index (P=.014) for the HA porous-coated Anatomic Porous Replacement component compared to the titanium Anatomic Porous Replacement component. Backscattered electron analysis showed 50% more bone in the HA porous-coated implant (P=.028). The mineral content analysis demonstrated that the bone ingrown into the HA porous-coated device was 23% less mineralized (P=.016). The data from this case study suggested that plasma-sprayed HA porous-coated implants may assist in increasing the amount of bone ingrowth and skeletal attachment in total hip arthroplasties.     

Lipid extraction enhances bank bone incorporation. An experiment in rabbits

We implanted frozen allogeneic cancellous bone in rabbit skeletal defects and compared the bone-forming response with that from similar implants that had also been extracted with chloroform/methanol. The donor bone was harvested from a previously implanted titanium chamber that is spontaneously filled with reproducible amounts of cancellous bone. It was processed as frozen bank bone, then transferred to an identical, but empty, chamber in another rabbit. Extraction of lipids before implantation increased the ingrowth of new bone into the transferred bone, as measured by 45Ca and 99mTc-MDP activity. A simple treatment with fat solvents may reduce some of the drawbacks of ordinary bank bone.