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[BMP-2 gene modified tissue-engineered bone repairing segmental tibial bone defects in goats].

OBJECTIVE: To evaluate the effectiveness of the tissue-engineered bone substitute loaded with adenovirus mediated human bone morphogenetic protein-2 gene (Adv-hBMP-2) transfected bone marrow derived mesenchymal stem cells (BMSC) in the repair of diaphyseal segmental bone defect of large animal. METHODS: The right tibial bone defects (2.6 cm) model of 26 goats were established and divided into 5 groups: I. Adv-hBMP-2 transfected BMSC/calcined bone (CB) group (n = 9); II. adenovirus-beta-galactosidase (Adv-betagal) gene transfected BMSC/CB group (n = 6); III. untransfected BMSC/CB group (n = 6); IV. single CB group (n = 3); VI. untreated group (n = 2). The above tissue-engineered bone substitutes were implanted in the bone defects respectively except group VI. Roentgenography, histomorphometrical analysis and biomechanical measurement were studied at various times. RESULTS: X-ray: at 4 - 8th weeks after implantation, more bony callus was found in the bone defects of group I. The complete healing rates of group I, II, III, IV, and V were 5/8, 1/5, 0/5, 0/2, 0/1 respectively at 26th week after implantation. Histomorphometrical analysis showed much more new bony callus including cortical bone formed in group I than those of other groups. The compression strength of the implanted bone substitute of group I is significantly higher than those of group II and III. CONCLUSION: The tissue-engineered bone substitute loaded with human BMP-2 gene transfected BMSC can repair diaphyseal segmental bone defect of large animal (goat).

Animals↗

[Clinical application of medical grade calcium sulfate as a bone graft substitute].

OBJECTIVE: To investigate the clinical effect of medical grade calcium sulfate(Osteoset) as a bone graft substitute. METHODS: From December 2004 to May 2005, 9 cases of bone defect(limb group) were repaired with Osteoset pellets; bone defect was caused by benign tumor in limbs, including 3 cases of fibroma xanthomas in humerus (1 case) and acetabulum (2 cases), 2 cases of bone cysts in humerus (1) and radius (1), 1 case of nonossifying fibroma, 1 case of ossifying fibroma and 2 cases of osteofibrous dysplasia in femurs. Five cases of lumbar posterolateral fusion (spine group) were treated with Osteoset pellets as autograft volume expander, including 2 cases of lumbar spinal stenosis, 2 cases of lumbar spondylolisthesis and 1 case of lumbar spondylolysis. Radiological method was used to evaluate the repair effect of Osteoset pellets. RESULTS: The mean follow-up time was 6.2 months (3 to 9 months). Osteoset pellets began to be absorbed after 1 to 3 months of operation, and were totally absorbed and replaced by osseous tissue after 4 to 6 months. No local recurrence was detected in limb group and the function of limbs was normal. At 6 months after operation, all patients in spine group got bony fusion. CONCLUSION: Medical grade calcium sulfate (Osteoset) is an ideal bone graft substitute with excellent bone repair effect.

Adolescent↗

Coralline hydroxyapatite bone graft substitute in hindfoot surgery.

BACKGROUND: Coralline hydroxyapatite bone graft substitute material is created by the conversion of the calcium carbonate structure of coral into hydroxyapatite. The microstructure of the material resembles cancellous bone and provides an osteoconductive scaffold for bone ingrowth. The use of this material is reported in the orthopedic oncology and trauma literature. Short-term reports in foot procedures have been reported. METHOD: A retrospective review identified 10 patients who had undergone hindfoot arthrodesis with coralline hydroxyapatite bone graft. The charts were reviewed for outcomes, operative complications, and time to union. Nine of the 10 patients were available for additional examination. RESULTS: The average followup was 6 years. There were three complications. One patient had sural nerve numbness with partial resolution which was unrelated to the graft material. A rheumatoid patient on methotrexate and prednisone developed a deep wound infection 9 months after surgery. One patient had a nonunion of the talonavicular joint. Eight of the 10 patients reported good or excellent results. Radiographs continued to demonstrate the presence of the graft material at 6 years. Radiographs showed extrusion of the graft from the joint in all patients. No patient had symptoms from the extruded material. DISCUSSION: Coralline hydroxyapatite bone graft substitute appears to be a clinically effective material for use in foot procedures. No adverse events could be linked to the graft material. The graft material is difficult to contain and extrusion was present in all patients. The slow resorption is a concerning characteristic of the graft material. At 6-year followup, the continued presence of the material has not shown any adverse effect.

Arthrodesis↗

In vivo evaluation of bone marrow stromal-derived osteoblasts-porous calcium phosphate ceramic composites as bone graft substitute for lumbar intervertebral spinal fusion.

STUDY DESIGN: Autogenous bone marrow stromal-derived osteoblasts-porous calcium phosphate ceramic composites were constructed in vitro under cell culture for 48 hours and implanted as a bone graft substitute for lumbar intervertebral spinal fusion in rabbits. OBJECTIVES: To evaluate the efficacy of autogenous bone marrow stromal-derived osteoblasts-porous calcium phosphate ceramic composites as an alternative to autogenous graft materials in a lumbar interbody spinal fusion model. SUMMARY OF BACKGROUND DATA: Bone marrow contains a population of rare progenitor cells capable of differentiating into bone, cartilage, muscle, tendon, and other connective tissues. These cells can be induced and differentiated into osteogenic osteoblasts with addition of osteogenic supplements. Combining bone marrow stromal-derived osteoblasts with porous ceramics gave rise to bone tissue in subcutaneous sites and repaired critical size segmental femoral defects. Little work has been done in the spine to assess fusion rates and associated biomechanical characteristics. METHODS: Five experimental groups were evaluated: sham operation (Group I); porous calcium phosphate ceramics alone (Group II); autogenous tricortical iliac crest (Group III); bone marrow stromal-derived osteoblasts-calcium phosphate ceramic composites (Group IV); bone marrow stromal-derived osteoblasts-calcium phosphate ceramic composites with rhBMP-2 (Group V). All rabbits were killed 12 weeks after surgery, and the spinal fusion segments underwent the evaluation of gross inspection, manual palpation, radiography, computed tomography, nondestructive biomechanical testing, and histologic analysis. RESULTS: Successful spinal fusion was achieved by manual palpation in 100% (6/6) of animals in Group IV and Group V, 66.7% (4/6) in Group III, 50% (3/6) in Group II, and 0% (0/6) in Group I. Radiographic studies showed that minimal disc height loss was observed with ceramic blocks than with autograft. Biomechanical testingconfirmed that spines from Group IV and Group V were statistically significantly stiffer in flexion, extension, left and right bending, and left and right torsion than Group III and Group II. Histologic analysis demonstrated a qualitative increase of bone formation in fusion mass in Group IV and Group V versus all other groups. The size of fusion mass and the stiffness of fusion segments were greatest in Group V. CONCLUSION: The results indicate that bone marrow stromal-derived osteoblasts-calcium phosphate ceramic composites may provide an alternative to autogenous graft materials for lumbar interbody spinal fusion. Adding recombinant human bone morphogenetic protein-2 into the composites may reinforce the biomechanical stiffness for spinal fusion segments. Porous calcium phosphate ceramics alone were not suitable as a bone graft substitute for lumbar interbody spinal fusion.

Animals↗

Resorption of synthetic porous hydroxyapatite and replacement by newly formed bone.

Synthetic porous hydroxyapatite (HA) is commonly used as a bone substitute for bone defects which, previously, would have been treated by autogenous bone grafting. HA has been thought to be a nonbiodegradable material that remains as it is implanted. However, after long-term follow-up, some authors report that the margin of implanted HA blocks or granules is absorbed, suggesting that HA is biodegradable. We experienced a patient in whom synthetic HA blocks implanted in a bone defect of the ilium after the harvesting of full-thickness bone for grafting were extensively absorbed and replaced by newly formed bone 6 years and 7 months after the implantation. Therefore, we conclude that HA is biodegradable. Sintering temperature, porosity, and pore diameter seem to influence the biodegradability of HA.

Absorption↗

Formation of a calcium phosphate-rich layer on absorbable calcium carbonate bone graft substitutes.

The use of natural coral as a bone graft substitute is common in Europe. However, the bone-coral bonding mechanism remains elusive. A rat subcutaneous model was used to demonstrate changes at the surface of resorbable calcium carbonate in the form of natural coral. Histological results indicated in vivo formation of a calcium phosphate (CaP)-rich layer on the surface of the coral confirmed by backscattered electron imaging and X-ray microanalysis. There appears to be a combination solution-mediated dissolution/cell-mediated degradation of the natural coral with subsequent surface conversion or precipitation. The end result is a CaP-rich layer on the coral. Though this layer has been observed previously, it was originally thought to be a histological artifact. This result is similar, however, to what is seen with Bioglass and glass ceramics and may also explain the good bonding of bone to hydroxyapatite. The fact that this layer is also present on natural coral after implantation in soft tissue sites may explain the intimate bone apposition observed when natural coral is placed in bony sites.

Absorption↗

Repair of experimental calvarial defects with Bio-Oss particles and collagen sponges in a rabbit model.

Various materials have been used for reconstruction of both acquired and congenital calvarial defects. Unfortunately, each has its limitations. Autologous bone grafts have irregular rates of resorption that may require secondary corrective surgery, and individual harvest sites have limited stores that can necessitate additional donor locations. Alloplastic materials have unlimited quantities and volume stability but they may not become incorporated and are associated with a higher incidence of infection. The optimal bone substitute should stimulate new bone formation and permanently supplant the temporary space filler, thereby reconstituting the surgical defect. We evaluated 2 newly available bone substitutes, resorbable natural bone mineral (Bio-Oss particles) and a combination of collagen and natural bone mineral collagen combination (Bio-Oss sponges), to repair calvarial defects in an adult, male, New Zealand white rabbit model. We found that the particulate Bio-Oss material resorbed and then underwent the normal physiological stages of bone remodeling. The collagen and Bio-Oss combination was replaced by new bone ingrowth. These materials may have potential for use in the reconstruction of skull defects.

Animals↗

Effect of rhBMP-2 on guided bone regeneration in humans.

The aim of the present clinical study was to test whether or not the addition of recombinant human bone morphogenetic protein-2 (rhBMP-2) to a xenogenic bone substitute mineral (Bio-Oss) will improve guided bone regeneration therapy regarding bone volume, density and maturation. In 11 partially edentulous patients, 34 Brånemark implants were placed at two different sites in the same jaw (five maxillae, six mandibles) requiring lateral ridge augmentation. The bone defects were randomly assigned to test and control treatments: the test and the control defects were both augmented with the xenogenic bone substitute and a resorbable collagen membrane (Bio-Gide). At the test sites, the xenogenic bone substitute mineral was coated with rhBMP-2 in a lyophilization process. Following implant insertion (baseline), the peri-implant bone defect height was measured from the implant shoulder to the first implant-bone contact. After an average healing period of 6 months (SD 0.17, range 5.7-6.2), the residual defects were again measured and trephine burs were used to take 22 bone biopsies from the augmented regions. The healing period was uneventful except for one implant site that showed a wound dehiscence, which spontaneously closed after 4 weeks. Later at reentry, all implants were stable. At baseline, the mean defect height was 7.0 mm (SD 2.67, range 3-12 mm) at test and 5.8 mm (SD 1.81, range 3-8 mm) at control sites. At reentry, the mean defect height decreased to 0.2 mm (SD 0.35, range 0-1 mm) at test sites (corresponding to 96% vertical defect fill) and to 0.4 mm (SD 0.66, range 0-2 mm) at the control site (vertical defect fill of 91%). Reduction in defect height from baseline to reentry for both test and control sites was statistically significant (Wilcoxon P<0.01). Histomorphometric analysis showed an average area density of 37% (SD 11.2, range 23-51%) newly formed bone at test sites and 30% (SD 8.9, range 18-43%) at control sites. The fraction of mineralized bone identified as mature lamellar bone amounted to 76% (SD 14.4, range 47.8-94%) at test compared to 56% (SD 18.3, range 31.6-91.4%) at control sites (paired t-test P<0.05). At BMP-treated sites 57% (SD 16.2, range 29-81%) and at control sites 30% (SD 22.6, range 0-66%) of the surface of the bone substitute particles were in direct contact with newly formed bone (paired t-test P<0.05). It is concluded that the combination of the xenogenic bone substitute mineral with rhBMP-2 can enhance the maturation process of bone regeneration and can increase the graft to bone contact in humans. rhBMP-2 has the potential to predictably improve and accelerate guided bone regeneration therapy.

Adult↗

Biological effects of calcium sulfate as a bone graft substitute in ovine metaphyseal defects.

Calcium sulfate has been used as a bone graft substitute in many fields, from dentistry to orthopedics. However, the results of many studies have yielded inconclusive results. In the present study, a sheep model was used with tibial and femoral metaphyseal defects to determine whether calcium sulfate was as effective as autograft and allograft in promoting new bone formation in a critical size defect. Medical-grade calcium sulfate pellets, autograft bone, allograft bone, or nothing was used to fill the metaphyseal defects. The sheep were allowed to heal for 12 weeks. Sagittal sections from the bones were analyzed with high-resolution contact radiographs, backscattered electron microscopy, and light microscopy. The volume fractions of bone within the defect perimeter were determined, and the histologic quality of the bone was observed. The volume fraction of new bone in the autograft, calcium sulfate, and allograft were not statistically different, but all were significantly different than the untreated control. The majority of the calcium sulfate had been resorbed at 12 weeks, and the histologic quality of the bone appeared similar to the autograft-treated bone. Calcium sulfate appears to be a useful biocompatible bone graft substitute that yields results similar to autograft bone in sheep metaphyseal defects.

Animals↗

Role of chitin beads in the formation of jaw bone by guided tissue regeneration. An experiment in the rat.

It has been reported that local application of bone grafts or synthetic bone substitutes (filler materials) may favour bone formation when used in combination with guided tissue regeneration (GTR). Therefore, the aim of the present investigation was to evaluate the effect of application of chitin beads (a bioabsorbable natural polymer) as a bone substitute in bone formation by GTR. The experiment was carried out in 25 rats. The mandibular ramus was exposed on one side after elevation of a muscle-periosteal flap, and a teflon capsule filled with chitin beads (2.0 mm in diameter) was placed with its opening facing the lateral aspect of the ramus. On the contralateral side of the jaw, serving as control, an empty teflon capsule was placed in the same manner. Groups of 5 animals were sacrificed at 7, 15, 30, 60 and 120 days following capsule placement. Histological analysis demonstrated that the amount of newly formed bone was similar in both experimental and control specimens, amounting to approximately 3% of the central/largest, cross-sectional area created by the capsule at 15 days, and to approximately 9% of this area at 30 days following capsule placement. At 60 and 120 days, however, the amount of newly formed bone observed in the control specimens was twice as large as that observed in the test specimens, amounting to approximately 31% of the cross-sectional area created by the capsule at 60 days, and to approximately 45% at 120 days. It is concluded that, although chitin beads (2.0 mm in diameter) are biocompatible, their presence retards bone formation in the model system used.

Absorbable Implants↗

The evaluation of processed cancellous bovine bone as a bone graft substitute.

OBJECTIVE: To determine the ability of a novel bovine cancellous bone xenoimplant to act as an osteoconductive graft in an ovine femoral defect model. An autograft harvested from the xenoimplant site was placed in a contralateral limb defect for comparison. MATERIAL AND METHODS: The xenoimplant used had been rendered immunologically inert by a novel defatting and deproteinating process. Following surgical implantation of cores into condylar cancellous bone defects, fluorochrome labels were administered to 12 sheep at 2 1/2, 4 1/2 and 8 weeks. Incorporation of the xenoimplants and autografts into the host bone was compared radiographically and histomorphometrically at 10 weeks. RESULTS: Radiographically, the degree of osteointegration was comparable. Histomorphometric data, consisting of labelled surface (LS) estimates, confirmed osteoconductive properties of both the xenoimplants and autografts. Remodelling activity was greatest in the xenoimplants at 2 1/2, weeks. At 4 1/2 weeks, there was more activity in the autograft, but by 8 weeks they were performing similarly. Xenoimplant-LS estimates were comparable or greater than those of the autograft at all times. CONCLUSIONS: Processed bovine cancellous bone xenoimplants were osteoconductive in this model and show promise for development as a biomaterial in human and veterinary orthopaedic surgery.

Animals↗

[Materials for reconstruction of the middle ear].

To rehabilitate most cases of conductive hearing loss closure of ear drum perforations and rebuilding of the ossicular chain can be performed. Due to the great number of biocompatible bone substitute materials available it is occasionally difficult for the surgeon to choose the most favorable substitute. Autogenous structures (ossicles, cortical bone, cartilage) and allogenous tissues (ossicles, cortical bone, cartilage, dentin) are possible bone replacement materials. Xenogenic tissue is currently not used in middle ear surgery. Ionomer cement is a hybrid material for replacement of bone but does not fit direct classification of the various classes of alloplastic materials in current use: that is, metals (gold, steel wire, platinum, titanium), plastics (polyethylene, polytetrafluorethylene) and ceramics (ceramic oxide, carbon, calcium-phosphate ceramic, vitreous ceramic). For restoration of the sound conductive apparatus preference is given to autogenous ossicles because cortical bone is resorbed and cartilage weakens over time. Most surgeons do not use allogenous tissue, because of the possible transmission of such infectious disease as immunodeficiency syndrome or Creutzfeldt-Jakob disease. Only dentin deserves special attention as a possible bone substitute in the middle ear because its form can be preserved during sterilization. Based on the observations available to date, it becomes apparent that titanium implants hold greater promise than gold. Form-stable synthetic materials are not generally recommended due to foreign body reactions which have been confirmed by many investigators. Ceramic materials (e.g. ceramic oxide, carbon, calcium-phosphate ceramic, glass ceramic) are well tolerated in the middle ear and have also proved to be useful over time. Hybrid bone substitute ionomer cement is easily workable and well integrated, showing a good functional outcome. For many years good results in otosclerosis surgery have been achieved with a prosthesis made of platinum-wire and Teflon. Short-term follow-up periods hold great promise with pistons made of gold. Autogenous ossicles, ionomer cement and recently titanium protheses--as far as usable--are employed by the author for reconstructing the middle ear. For the time being platinum-Teflon prostheses and gold are used in otosclerosis surgery.

Animals↗

In vivo evaluation of resorbable bone graft substitutes in a rabbit tibial defect model.

Calcium sulfate as a bone graft substitute is rapidly resorbed in vivo releasing calcium ions but fails to provide long-term three-dimensional framework to support osteoconduction. The setting properties of calcium sulfate however allow it to be applied in a slurry form making it easier to handle and apply in different situations. This study examines the in vivo response of calcium sulfate alone and as a carrier for a coralline hydroxyapatite in an established bilateral corticocancellous defect model in rabbits. Defects were filled flush to the anterior cortex with a resorbable porous ceramic alone and in combination with calcium sulfate slurry, calcium sulfate slurry alone or calcium sulfate pellets and examined at time points up to 52 weeks. Specimens where assessed using Faxitron X-ray, light and electron microscopy. Calcium sulfate in either slurry or pellet form does indeed support new bone formation alone however, complete filling of the bone defect is not observed. Calcium sulfate in slurry form does however improve the surgical handling of particulate bone graft substitutes such as Pro Osteon 200 R, which remained as an osteoconductive scaffold for up to 52 weeks and may have played an important role in the ultimate closure of the cortical windows.

Absorbable Implants↗

Developing porosity of poly(propylene glycol-co-fumaric acid) bone graft substitutes and the effect on osteointegration: a preliminary histology study in rats.

Bioresorbable bone graft substitutes could eliminate disadvantages associated with the use of autografts, allografts and other synthetic materials. We investigated a bioresorbable bone graft substitute made from the unsaturated polyester poly(propylene fumarate) which is crosslinked in the presence of soluble and insoluble calcium filler salts. This compact bone graft substitute material develops porosity in vivo by leaching of the soluble filler salts. In attempt to develop materials whose in vivo porosity can be designed such that implant degradation would occur at a rate that remains supportive of the overall structural integrity of the repairing defect site, we studied the early tissue response upon implantation in a bony defect. Three grout formulations of varying solubilities using slightly soluble hydroxyapatite (HA) and soluble calcium acetate (CA) were evaluated in 3 mm holes made in the anteromedial tibial metaphysis of 200 g Sprague Dawley rats (n = 16 per formulation for a total of 48 animals). Grout formulations cured in situ. Animals from each formulation were sacrificed in groups of 8 at 4 days and 3 weeks postoperatively. Histologic analysis of the healing process revealed improved in vivo osteointegration of bone graft substitutes when a higher loading of calcium acetate was employed. All formulations maintained implant integrity and did not provoke sustained inflammatory responses. This study suggested that the presence of a soluble salt permits in vivo development of porosity of a poly(propylene fumarate) based bone graft substitute material.

Acetates↗

[Development of artificial bone made from nacre and polylactic acid and property assessment].

OBJECTIVE: To explore a new bone substitute for bone grafting and evaluate its properties. METHODS: Pressurized molding of nacre, polylactic acid and NaCl that were proportionally mixed was performed to obtain the bone grafting material, and its properties as porosity, pore size and mechanical strength were compared with those of the material prepared by solvent casting. RESULTS: Compared with that of the material made by solvent casting, the mechanical strength of the new artificial bone material was 10-fold stronger with an average porosity rate of 47.65%, pore size of 218.83 micrometer compressive strength of 16.10 MPa and flexural strength of 60.18 MPa. CONCLUSION: The new technique adopted in this study can effectively enhance the biomechanics of the artificial bone material, which can be suitable for repairing bone defect.

Biocompatible Materials↗

[The experimental study on the activity of rhBMP-2, coral and collagen composites inducing intramuscle bone].

OBJECTIVE: To investigate the ectopic bone induction activity of the composite of coral, collagen and rhBMP-2, and the degradation of the carriers of rhBMP-2. METHODS: The composites of rhBMP-2, coral and collagen were implanted into the muscle pouches of mice. The mice were divided into the following four groups. Group H: each composite had 2 mg rhBMP-2, Group L: each composite had 0.5 mg rhBMP-2, Group J: each composite had only type I collagen, Group C: only coral was implanted in the muscle of mouse. Each group had 20 mice. Every 5 mice in each group were killed postoperatively after 1, 2, 4 and 8 weeks. The ectopic osteoinductive activity was assessed with histology and histomorphometry. The point counting method was applied to measure the formation of new bone and cartilage and the degradation of coral in the composites. The data was analyzed statistically using t test. RESULTS: There was no obvious morbidity in any of the animals in this study, and no wound infection, bleeding, or feeding difficulties postoperatively. The samples were examined histologically. Cartilage and bone induction occurred in Group H and L. After 1 Week, new bone was observed in Group H, and cartilage formation occurred mainly in Group L. After 2 weeks, both cancellous bone and bone marrow elements were present in Group H and L. New bone formation was not uniform throughout the specimen. Part of the coral in the composites was not absorbed between 2 to 4 weeks postoperatively. At the 8th week, the coral in the composite was absorbed completely. While in Group J and C there was not any new cartilage or bone formation. According to the statistical analysis, bone formation occurred 2 weeks earlier in Group H than in Group L after operation. From 4 postoperative weeks, no difference of the bone formation rate was observed between Group H and Group L. The degradation rate of coral in Group C was the fastest among the four groups. CONCLUSION: The composites of rhBMP-2, coral and collagen are degradable bone substitutes with bone induction and conduction, and induce dosedependent and time-dependent amounts of intramuscle bone in mice during limited postoperative period. The coral in the composites could be absorbed completely.

Animals↗

Guided bone regeneration around exposed implants: a new bioresorbable device and bioresorbable membrane pins.

This article presents the principles and the clinical procedure of utilizing a new bioresorbable collagen barrier device, bioresorbable membrane pins, and a new bone substitute for bone regeneration around exposed implants. Three cases are used to illustrate the clinical procedure. In all three cases, the regeneration of bone around the implants was successful, and upon clinical examination the newly regenerated material resembled natural bone. Histologic data regarding the new bioresorbable collagen barrier itself and the time range in which the collagen device functioned as a barrier are still lacking, and the studies are currently being conducted. The learning objective of this article is to introduce a combination of options available for successful regeneration of bone around the implants and avoiding the second-stage surgery.

Adult↗

Acetabular reconstruction in total hip revision using a bone graft substitute. Early clinical and radiographic results.

Thirty revisions were performed on loose total hip replacements with a specific acetabular reconstruction technique combining the use of a bovine bone substitute and a support ring. One patient died of a cause unrelated to the procedure. No migration of the acetabular implant or bone lysis of the heterograft was seen in 27 cases after 3 years. Radiologically, the heterograft gradually condensed, and its appearance matched that observed with an allograft. The 2 failures with implant migration and heterograft lysis were interpreted as technical errors related to the use of the Müller ring. In both cases the Müller ring was supported by the cancellous heterograft and not by the host bone.

Acetabulum↗