Aims. To clarify the mid-term results of transposition osteotomy of the acetabulum (TOA), a type of spherical periacetabular osteotomy, combined with structural allograft bone grafting for severe hip dysplasia. Methods. We reviewed patients with severe hip dysplasia, defined as Severin IVb or V (lateral centre-edge angle (LCEA) < 0°), who underwent TOA with a structural bone allograft between 1998 and 2019. A medical chart review was conducted to extract demographic data, complications related to the osteotomy, and modified Harris Hip Score (mHHS). Radiological parameters of hip dysplasia were measured on pre- and postoperative radiographs. The cumulative probability of TOA failure (progression to Tönnis grade 3 or conversion to total hip arthroplasty) was estimated using the Kaplan–Meier product-limited method, and a multivariate Cox proportional hazard model was used to identify predictors for failure. Results. A total of 64 patients (76 hips) were included in this study. The median follow-up period was ten years (interquartile range (IQR) five to 14). The median mHHS improved from 67 (IQR 56 to 80) preoperatively to 96 (IQR 85 to 97) at the latest follow-up (p < 0.001). The radiological parameters improved postoperatively (p < 0.001), with the resulting parameters falling within the normal range in 42% to 95% of hips. The survival rate was 95% at ten years and 80% at 15 years. Preoperative Tönnis grade 2 was an independent risk factor for TOA failure. Conclusion. Our findings suggest that TOA with structural bone allografting is a viable surgical option for correcting severely dysplastic acetabulum in adolescents and young adults without advanced osteoarthritis, with favourable mid-term outcomes. Cite this article: Bone Joint J 2023;105-B(7):743–750

Bone allografts can be used in any kind of surgery involving bone from minor defects to major bone loss after tumour resection. This review describes the various types of bone grafts and the current knowledge on bone allografts, from procurement and preparation to implantation. The surgical conditions for optimising the incorporation of bone are outlined, and surgeon expectations from a bone allograft discussed

Bone infections due to fractures or implants are a big medical problem. In experimental medicine, many experimental models have been created on different animal species to simulate the disease condition and to do experience treatments. The aim of this paper was to present an antibacterial efficacy of using a bone allograft developed according to the Marburg system of bone bank on a model of chronic osteomyelitis induced in rabbits. In research was used 54 rabbits. Osteomyelitis was induced in rabbits by a human strain of St. aureus ATCC 43300, in the rabbit femur. There have been created 3 groups of animals. In 1. st. group used antibiotic impregnated biodegradable material “PerOssal”. In 2. nd. group used antibiotic impregnated whole bone allograft. In 3. rd. group used antibiotic impregnated perforated bone allograft. Evaluation of installation and evolution of the disease was done by microbiological. A separate study of microbiological data is presented here. This study showed, in the 1. st. and 3. rd. groups there is a persistent decrease in CFU by 14 knocks to 120.4 in the 1. st. group and to 3.5 in the 3. rd. group, and in the 2. nd. group, on the contrary, there is an increase in CFU to 237.33. This shows the lack of effectiveness of using a whole bone allograft. The results showed, after 7 days there was no statistically significant difference between the groups. After 14 days the perforated bone allograft impregnated with antibiotic was better than the biodegradable material “PerOssal”

Introduction. Cancellous and cortical bone used as a delivery vehicle for antibiotics. Recent studies with cancellous bone as an antibiotic carrier in vitro and in vivo showed high initial peak concentrations of antibiotics in the surrounding medium. However, high concentrations of antibiotics can substantially reduce osteoblast replication and even cause cell death. Objectives. To determine whether impregnation with gentamycine impair the incorporation of bone allografts, as compared to allografts without antibiotic. Materials and method. Seventy two healthy rabbits (24 rabbits in each group) were used for this study. Bone defects (3-mm diameter, 10-mm depth) were created in the femur. Human femoral head prepared according to the Marburg bone bank system was used as bone allograft. In the experimental groups, in 1 group - the defects were filled with bone allografts, in 2 group – Perforated Gentamycin-impregnated bone allografts. The control group did not receive any filling. The animals were killed after 14, 30 and 60 days. Evaluations consisted of X-ray plain radiography, histology at 14-, 30- and 60-days post-surgery. Results. Active osteoblast activity and active formation of new bones were detected around the defect area in all groups, but the amount of new bone formation was greater in the experimental groups than the control group. We found no statistically significant differences in the rate of bone formation between 1 and 2 groups at 14, 30 and 60 days in any of the parameters studied. X-ray results showed no significant difference in bony callus formation around allografts in 1 and 2 groups. In contrast, no significant callus formation was observed in the control group. Conclusion. The use of gentamycin-impregnated bone allografts may be of value in procedures performed at the site of osteomyelitis which require a second stage reconstruction with impacted bone grafting techniques

Bone allograft use in trauma and orthopaedic surgery is limited by the potential for cross infection due to inadequate acceptable decontamination methods. Current methods for allograft decontamination either put the recipient at risk of potentially pathogenic organisms or markedly reduce the mechanical strength and biological properties of bone. This study developed a technique of sterilization of donor bone which also maintains its mechanical properties. Whole mature rat femurs were studied, as analogous to strut allograft. Bones were inoculated by vortexing in a solution of pathogens likely to cause cross infection in the human bone graft situation. Inoculated bones were subjected to supercritical carbon dioxide at 250 bar pressure at 35 degrees celsius for different experimental time periods until a set of conditions for sterilization was achieved. Decontamination was assessed by vortexing the treated bone in culture broth and plating this on suitable culture medium for 24 hours. The broth was also subcultured. Controls were untreated-, gamma irradiated- and dehydrated bone. Mechanical testing of the bones by precision three-point bending to failure was performed and the dimensions and cross-section digitally assessed so values could be expressed in terms of stress. Mechanical testing revealed bone treated with supercritical carbon dioxide was consistently significantly stronger than that subjected to gamma irradiation and bones having no treatment (due to the minor dehydrating effect of the carbon dioxide). Terminal sterilization of bone is achieved using supercritical carbon dioxide and this method maintains the mechanical properties. The new technique greatly enhances potential for bone allograft in orthopaedic surgery

We retrospectively reviewed 40 hips in 36 patients who had undergone acetabular reconstruction using a titanium Kerboull-type acetabular reinforcement device with bone allografts between May 2001 and April 2006. Impacted bone allografts were used for the management of American Academy of Orthopaedic Surgeons Type II defects in 17 hips, and bulk bone allografts together with impacted allografts were used for the management of Type III defects in 23 hips. A total of five hips showed radiological failure at a mean follow-up of 6.7 years (4.5 to 9.3), two of which were infected. The mean pre-operative Merle d’Aubigné score was 10 (5 to 15) vs 13.6 (9 to 18) at the latest follow-up. The Kaplan-Meier survival rate at ten years, calculated using radiological failure or revision of the acetabular component for any reason as the endpoint, was 87% (95% confidence interval 76.3 to 97.7). A separate experimental analysis of the mechanical properties of the device and the load-displacement properties of bone grafts showed that a structurally hard allograft resected from femoral heads of patients with osteoarthritis should be preferentially used in any type of defect. If impacted bone allografts were used, a bone graft thickness of < 25 mm was acceptable in Type II defects. This clinical study indicates that revision total hip replacement using the Kerboull-type acetabular reinforcement device with bone allografts yielded satisfactory mid-term results

Aims. The aim of this study was to report the medium-term outcomes of impaction bone allograft and fibular grafting for osteonecrosis of the femoral head (ONFH) and to define the optimal indications. Methods. A total of 67 patients (77 hips) with ONFH were enrolled in a single centre retrospective review. Success of the procedure was assessed using the Harris Hip Score (HHS) and rate of revision to total hip arthroplasty (THA). Risk factors were studied, including age, aetiology, duration of hip pain, as well as two classification systems (Association Research Circulation Osseous (ARCO) and Japanese Investigation Committee (JIC) systems). Results. After a mean follow-up period of 8.61 years (SD 1.45), 81.3% (52/64) of enrolled cases had a good or excellent HHS at latest follow-up (declining to 76.0% (38/50) for those with more than eight years of follow-up). Overall survival was 92.1% at eight years’ follow-up (95% CI 83.2% to 96.4%). A total of 12 hips (19.0%) failed (reoperation or HHS < 70 points) at final follow-up. Rate of success was adversely affected by increasing age, duration of pain, and more severe disease as measured using the ARCO and JIC classifications, but not by aetiology of the ONFH. Conclusion. We report favourable medium-term results of this procedure. Best outcomes can be expected in patients matching the following indications: younger than 40 years; less 12-month hip pain before surgery; femoral head collapse being less than 2 mm; and integrated lateral wall of femoral head. Cite this article: Bone Joint J 2020;102-B(7):838–844

The purpose of this study is to enhance massive bone allografts osseointegration used to reconstruct large bone defects. These allografts show >50% complication rate requiring surgical revision in 20% cases. A new protocol for total bone decellularisation exploiting the vasculature can offer a reduction of postoperative complication by annihilating immune response and improving cellular colonization/ osseointegration. The nutrient artery of 18 porcine bones - humerus/femur/radius/ulna - was cannulated. The decellularization process involved immersion and sequential perfusion with specific solvents over a course of one week. Perfusion was realized by a peristaltic pump (mean flow rate: 6ml/min). The benefit of arterial perfusion was compared to a control group kept in immersion baths without perfusion. Bone samples were processed for histology (HE, Masson's trichrome and DAPI for cell detection), immunohistochemistry (IHC : Collagen IV/elastin for intraosseous vascular system evaluation, Swine Leukocyte Antigen – SLA for immunogenicity in addition to cellular clearance) and DNA quantification. Sterility and solvent residues in the graft were also evaluated with thioglycolate test and pH test respectively. Compared to native bones, no cells could be detected and residual DNA was <50ng/mg dry weight. Intramedullary spaces were completely cleaned. IHC showed the preservation of intracortical vasculature with channels bounded by Collagen IV and elastin within Haversian systems. IHC also showed a significant decrease in SLA signaling. All grafts were sterile at the last decellularization step and showed no solvent residue. The control group kept in immersion baths, paired with 6 perfused radii/ulnae, showed that the perfusion is mandatory to ensure complete decellularisation. Our results prove the effectiveness of a new concept of total bone decellularisation by perfusion. These promising results could lead to a new technique of Vascularized Composite Allograft transposable to pre-clinical and clinical models

Bone allograft is the most widely accepted approach in treating patients suffering from large segmental bone defect regardless of the advancement of synthetic bone substitutes. However, the long-term complications of allograft application in term of delayed union and nonunion were reported due to the stringent sterilization process. Our previous studies demonstrated that the incorporation of magnesium ions (Mg2+) into biomaterials could significantly promote the gene up-regulation of osteoblasts and new bone formation in animal model. Hence, our group has proposed to establish an Mg2+ enriched tissue microenvironment onto bone allograft so as to enhance the bone healing. The decellularization and gamma irradiation process were performed on bovine bone allograft and followed by magnesium plasma treatment. To evaluate the biocompatibility and bioactivity, materials characterizations, in vitro and in vivo studies were conducted, respectively. Mg composite layer on bone surface ranged from 500nm to ∼800nm thick. The cell viability on magnesium enriched allograft was significantly higher than that of the control. The ALP gene expression of hTMSCs in the group of PIII&D treated samples was highly up-regulated. The bone regeneration ability of Mg modified bone allograft implanted in animal model was significantly superior than the control after 2-month post-operation

Bone allografts can store and release high levels of vancomycin. We present our results of a two-stage treatment for infected hip arthroplasty with acetabular and femoral impaction grafting using vancomycin-loaded allografts. We treated 29 patients (30 hips) by removal of the implants, meticulous debridement, parenteral antibiotic therapy and second-stage reconstruction using vancomycin-supplemented impacted bone allografts and a standard cemented Charnley femoral component. The mean follow-up was 32.4 months (24 to 60). Infection control was obtained in 29 cases (re-infection rate of 3.3%; 95% confidence interval 0.08 to 17) without evidence of progressive radiolucent lines, demarcation or graft resorption. One patient had a further infection ten months after revision caused by a different pathogen. Associated post-operative complications were one traumatic periprosthetic fracture at 14 months, a single dislocation in two hips and four displacements of the greater trochanter. Vancomycin-supplemented allografts restored bone stock and provided sound fixation with a low incidence of further infection

Impacted bone allograft is often used in revision joint replacement. Hydroxyapatite granules have been suggested as a substitute or to enhance morcellised bone allograft. We hypothesised that adding osteogenic protein-1 to a composite of bone allograft and non-resorbable hydroxyapatite granules (ProOsteon) would improve the incorporation of bone and implant fixation. We also compared the response to using ProOsteon alone against bone allograft used in isolation. We implanted two non-weight-bearing hydroxyapatite-coated implants into each proximal humerus of six dogs, with each implant surrounded by a concentric 3 mm gap. These gaps were randomly allocated to four different procedures in each dog: 1) bone allograft used on its own; 2) ProOsteon used on its own; 3) allograft and ProOsteon used together; or 4) allograft and ProOsteon with the addition of osteogenic protein-1. After three weeks osteogenic protein-1 increased bone formation and the energy absorption of implants grafted with allograft and ProOsteon. A composite of allograft, ProOsteon and osteogenic protein-1 was comparable, but not superior to, allograft used on its own. ProOsteon alone cannot be recommended as a substitute for allograft around non-cemented implants, but should be used to extend the volume of the graft, preferably with the addition of a growth factor

Successful reconstructive surgery with allografts is severely limited by a failure rate of 30 – 40%. Allograft failure is due to nonunion of the graft-host junction. The molecular mechanism by which this occurs is not yet fully elucidated. Using a sheep femoral allograft model, we have investigated the cellular and molecular mechanisms associated with nonunion of bone allografts. Five, from a total of twelve operations, resulted in the development of graft-host nonunion, reflecting a failure rate of 42%. Histological assessment revealed that allograft failure was due to the excessive accumulation of and resorption by, osteoclasts (Ocs) on the surface of the bone allograft. Three distinct layers, lying adjacent to the allograft bone surface, in the nonunion groups, were identified. The outer fibroblastic layer contained abundant fibroblasts and connective tissue. Underlying this layer were synovial-like cells and some multinuclear giant cells. The third layer, opposing the bone surface, consisted of Ocs and round mononuclear cells. Histomorphometric analysis showed that allograft unions, featured a large amount of newly formed bone on the surface, (OS/BS = 47.81%) with a small proportion of eroded surface (ES/BS = 20.59%). The number of osteoclasts associated with the allograft bone surface were few (Oc/B.Pm = 1.7190/mm) and activity (ES/BS = 46.68%) of Ocs with a reduced amount of new bone formation (OS = 6.35%). Both calcitonin receptor and H+ATPase mRNA, characteristically expressed by Ocs, were localised to the multinuclear giant cells, indicating that they were Ocs. Synovial-like cells in the histological layer above the Ocs, expressed gene transcript for the Osteoprotegrin Ligand (OPGL), a membrane bound factor that is critical for the induction of Oc activity and osteoclastogenesis. In conclusion, these findings suggest that failure of bone allografts is partially due to excessive resorption by host Ocs, accompanied by reduced bone formation. The production of OPGL by synovial-like cells, may be responsible for the recruitment and generation of Ocs

The Use of bone allograft in orthopaedic Surgery has been predicted to increase particularly in joint revision surgery. This has led to a potential problem with supply. Questionnaires were distributed to all 146 Consultant orthopaedic Surgeons working in Scotland in 2000. They were asked to indicate their current usage of bone and tissue allograft, any problems encountered with supply and if alternatives to allograft such as processed bone, might be used. The questions asked were very similar to those asked in a survey by the author (GG) in 1995 to enable comparisons to be made. 74% of all bone issued by the SNBTS in 2000 –2001 was used in revision hip arthroplasy. This compares with only 66% of bone issued in 1998–1999. Replies were received from 125 consultants (87%) of whom 93 reported using bone allograft. 41 consultants (46%) predicted an increase in their requirement for bone allograft, and 23 (26%) felt they could currently use more bone if this became available. Sixty percent of Surgeons would consider using processed bone as an alternative. In comparison with figures from a previous study in 1995, an increasing number of surgeons are prepared to use processed bone as an alternative to fresh frozen allograft. As the number of revision THR’ s continues to increase the amount of bone required is likely to continue to increase. The need to increase efficiency in harvest and supply of bone is therefore great. The use of more SNBTS nurses in selection of patients and collection of bone may increase efficiency. More surgeons may have to use processed bone, which would allow more bone to be released. Also processing may help reduce transmission of infective particles such as HIV and new variant CJD. With rising public and medical concerns over these issues this seems most desirable

Major drawbacks associated with autologous bone grafting are the risk of donor site morbidity and its limited availability. Sterilized bone allograft, however, lacking osteoinductive properties, carries the risk of graft failure resulting from insufficient osseointegration of the graft. The aim of this study was to vitalize bone allograft with human osteoprogenitor cells under GMP-conform conditions. For this purpose we investigated proliferation, osteogenic differentiation and large-scale gene expression of human MSCs cultured three-dimensionally on peracetic acid (PAA)-treated spongious bone chips. MSCs were isolated from healthy donors (N=5) and seeded onto PAA-treated spongious bone samples (~5×5×5 mm, DIZG, Germany) under GMP-conform conditions. Proliferation (total protein assay), osteogenic differentiation (cell-specific ALP activity assay, quantitative gene expression analysis of selected osteogenic marker genes), and morphology were assessed. RNA was isolated and microarray analysis was performed using the PIQORTM Stem Cell Microarray system (Miltenyi Biotec) including 942 target sequences. Increasing cellularity was observed during the 42 d observation period while cell-specific ALP activity peaked at day 21. Effective proliferation and adhesion of human MSCs on PAA-treated spongious bone was confirmed by histology, scanning electron and confocal laser scanning microscopy. Gene expression of early (Runx-2), intermediate (ALP), and late (osteocalcin) osteogenic marker genes was present during 42 days of cultivation. Microarray analysis of MSCs cultivated on bone allograft versus 2-D tissue culture demonstrated temporal upregulation of genes involved in extracellular matrix synthesis (e.g., matrix metalloproteases, collagens), osteogenesis (e.g., BMPR1b, Runx-2) and angiogenesis (angiopoietin, VEGF). PAA-treated spongious bone allograft is a biocompatible carrier matrix for long-term ex vivo cultivation of MSCs as observed by favorable proliferation, cell distribution, gene expression profile, and persisting osteogenic differentiation. GMP-grade vitalisation of bone allograft by cultivation with autologous MSCs represents a promising clinical application for the treatment of osseous defects

Metal meshes are used in revision surgery of the hip to contain impacted bone grafts in cases with cortical or calcar defects in order to provide rotational stability to the stem. However, the viability of bone allografts under these metal meshes has been uncertain. We describe the histological appearances of biopsies obtained from impacted bone allografts to the calcar contained by a metal mesh in two femoral reconstructions which needed further surgery at 24 and 33 months after the revision procedure. A line of osteoid and viable new bone was observed on the surface of necrotic trabeculae. Active bone marrow between these trabeculae showed necrotic areas in some medullary spaces with reparative fibrous tissue and isolated reactive lymphocytes. This is interpreted as reparative changes after revascularisation of the cancellous allografts. These pathological findings are similar to those reported in allografts contained by cortical host bone and support the hypothesis that incorporation of morcellised bone under metal meshes is not affected by these devices

Vancomycin-supplemented allografts provide biological restoration of bone stock and sound fixation with a low incidence of re-infection. Experimental incorporation of these grafts is similar to allografts without vancomycin. However, the underlying biology remains unknown. We report the first histological observations of vancomycin-supplemented impacted bone allografts in two reconstructions performed 14 and 20 months after revision surgery because of a periprosthetic fracture. Areas of active bone remodelling (creeping substitution), as well as calcified bone trabeculae and graft particles embedded in dense fibrous tissue, were observed with osteoid and fibroconnective tissue surrounding polymethylmethacrylate particles. These pathological findings are similar to those reported in allografts without vancomycin and support the hypothesis that high levels of vancomycin do not affect the incorporation of bone graft

Background: Early failure of morselized impaction bone allograft is usually due to shear forces. Soil mechanics tells us that in aggregates such as bone allograft, the resistance to these shear forces can be increased by altering the fluid concentration, varying the particle size, and improving the morphology of the graft particle. Finding an idealized concentration of fat and water in bone graft could improve the resistance to interparticle shear and therefore decrease the failure rate of impaction bone graft. Ensuring the quality of the bone source is adequate can also improve the initial strength the bone graft. Furthermore optimizing the graft can be achieved by screening methods and simple intra-operative techniques. Methods: Human femoral heads were retrieved from both total hip arthroplasty and hemiarthroplasty procedures. Bone mineral density was determined by DEXA scanning. The fat and water content of the graft was varied by combinations of squeezing and drying the graft and also by washing the graft using pulse lavaged water and 1:1 mixture of chloroform: methanol. The amount and characteristics of the fat and water in human morselized cancellous bone was quantified by the Karl Fischer extraction techniques, and gas chromatography. The overall shear strength of each graft preparation was determined by the direct shear test, adapted from an accepted protocol in soil mechanics and the optimum mixture which would resist shear forces was determined. Results: An optimum level of fat and water was determined which was 50% stronger than unaltered bone graft. This is most closely approximated in an operating theatre situation by washing the graft with pulse lavaged normal saline and subsequently squeezing the bone graft in a vice with a force of 335kPa for 5 minutes. Whereas osteoarthritic and osteoporotic bone were similar in their fat and water content and initial resistance to shear forces, after processing, the resistance to shear forces of osteoarthritic bone improved by 147% and that of osteoporotic bone only improved by 12% (p< 0.001) Conclusions: Optimizing the fat and water content of bone graft and closely choosing the source of graft produces a stronger graft which is more resistance to shear stresses, protecting the surgical construct until bone growth can occur

Management of bone defects is a common surgical challenge encountered following any high energy trauma. Femur fractures with bone loss account for 22% of all the fractures with bone loss/defect, and 5% to 10% of distal femur fractures are open injuries. It was estimated in 2008, that, more than 4.5 million open fractures occur annually in India. In this retrospective study, patients who received bone allograft from our tissue bank between May 2012 and September 2015 were analysed. Of the 553 allografts issued, at that point in time, 26 were used in patients who underwent reconstruction for distal Femur fractures primarily. Fractures with defect or bone loss from 12 cc (1cm) to 144 cc (12cm) were treated with either Internal or External fixation and bone allograft. Morcellised cancellous, or a cortical strut, were used to fill or reconstruct the defect or void. The radiological outcome in terms of fracture union was assessed and Knee society score was used to assess the functional outcome. Complications such as non- union, infection, stiffness and need of revision or additional procedures were also assessed. Osseous consolidation was achieved in all the 26 patients with a Median time of 24 weeks (16 to 60). The Median Functional Knee Society Score was 80, indicating satisfactory functional outcome. Infection was noted in one patient, but it was not attributed to the allograft. Additional minor procedures like bone marrow infiltration, corticotomy for bone lengthening were required in 10 patients. Our studycomprises the largest group of patients treated primarily with Allograft to reconstruct or fill the void of bone loss encountered with distal Femur fracture. Reconstruction of massive bone defects, in patients of distal Femur fractures, with bone allograft, shows encouraging results. The surgeon can achieve the goal of restoring form and function of these difficult injuries in a single stage and the technique will provide the freedom to reconstruct the bony defect up to 150 cc (12 cm length) and recreate the anatomy to near normal. This allows for early mobilisation of patients and restoration of their daily routine at the earliest

Structural bone allografts are a viable option in reconstructing massive bone defects in patients following musculoskeletal (MSK) tumour resection and revision hip/knee replacements. To decrease infection risk, bone allografts are often sterilised with gamma-irradiation, which consequently degrades the bone collagen connectivity and makes the bone brittle. Clinically, irradiated bone allografts fracture at rates twice that of fresh non-irradiated allografts. Our lab has developed a method that protects the bone collagen connectivity through ribose pre-treatment while still undergoing gamma-irradiation. Biomechanical testing of bone pretreated with our method provided 60–70% protection of toughness and 100% protection of strength otherwise lost with conventional irradiation. This study aimed to determine if the ribose-treated bone allografts are biocompatible with host bone. The New Zealand White rabbit (NZWr) radius segmental defect model was used, in which 15-mm critically-sized defects were created. Bone allografts were first harvested from the radial diaphysis of donor female NZWr, and treated to create 3 graft types: C=untreated controls, I=conventionally-irradiated (33 kGy), R=our ribose pretreated + irradiation method. Recipient female NZWr (n=24) were then evenly randomised into the 3 graft groups. Allografts were surgically fixed with a 0.8-mm Kirschner wire. Post-operative X-rays were taken at 2, 6, and 12 weeks, with bony healing assessed by a blinded MSK radiologist using an established radiographic scoring system. The reconstructed radii were retrieved at 12 weeks and analysed using bone histomorphometry and microCT. Kruskal-Wallis and Mann-Whitney tests were utilised to compare groups, with statistical significance when p<0.05. Radiographic analysis revealed no differences in periosteal reaction and degree of osteotomy site union between the groups at any time point. Less cortical remodeling was observed in R and I grafts compared to untreated controls at 6 weeks (p=0.004), but was no longer evident by 12 weeks. Radiographic union was achieved in all groups by 12 weeks. Histologic and microCT analysis further confirmed union at the graft-host bone interface, with the presence of mineralising callus and osteoid. Histomorphometry also showed the bridging external callus originated from host bone periosteum and a distinct cement line between allograft and host bone was present at the union site. Previous studies have shown that the presence of non-enzymatic glycation end products in bone can impair fracture healing. However, these studies investigated bony healing in the setting of diabetic states. Our findings showed that under normal conditions, ribose pretreated grafts healed at rates similar to controls via mechanisms also seen in retrieved human allografts clinically in use. These findings that grafts pretreated with our method are biocompatible with host bone in the rabbit help to further advance this technology for clinical trials

We reviewed the clinical and radiological results of 131 patients who underwent acetabular revision for aseptic loosening with impacted bone allograft and a cemented acetabular component. The mean follow-up was 51.7 months (24 to 156). The mean post-operative Merle D’Aubigné and Postel scores were 5.7 points (4 to 6) for pain, 5.2 (3 to 6) for gait and 4.5 (2 to 6) for mobility. Radiological evaluation revealed migration greater than 5 mm in four acetabular components. Radiological failure matched clinical failure. Asymptomatic radiolucent lines were observed in 31 of 426 areas assessed (7%). Further revision was required in six patients (4.5%), this was due to infection in three and mechanical failure in three. The survival rate for the reconstruction was 95.8% (95% confidence interval 92.3 to 99.1) overall, and 98%, excluding revision due to sepsis. Our study, from an independent centre, has reproduced the results of the originators of the method

We analysed the histological findings in 1146 osteoarthritic femoral heads which would have been considered suitable for bone-bank donation to determine whether pathological lesions, other than osteoarthritis, were present. We found that 91 femoral heads (8%) showed evidence of disease. The most common conditions noted were chondrocalcinosis (63 cases), avascular necrosis (13), osteomas (6) and malignant tumours (one case of low-grade chondrosarcoma and two of well-differentiated lymphocytic lymphoma). There were two with metabolic bone disease (Paget’s disease and hyperparathyroid bone disease) and four with inflammatory (rheumatoid-like) arthritis. Our findings indicate that occult pathological conditions are common and it is recommended that histological examination of this regularly used source of bone allograft should be included as part of the screening protocol for bone-bank collection

We analysed the histological findings in 1146 osteoarthritic femoral heads which would have been considered suitable for bone-bank donation to determine whether pathological lesions, other than osteoarthritis, were present. We found that 91 femoral heads (8%) showed evidence of disease. The most common conditions noted were chondrocalcinosis (63 cases), avascular necrosis (13), osteomas (6) and malignant tumours (one case of low-grade chondrosarcoma and two of well-differentiated lymphocytic lymphoma). There were two with metabolic bone disease (Paget’s disease and hyperparathyroid bone disease) and four with inflammatory (rheumatoid-like) arthritis. Our findings indicate that occult pathological conditions are common and it is recommended that histological examination of this regularly used source of bone allograft should be included as part of the screening protocol for bone-bank collection

Introduction: Bone grafts are frequently used in orthopaedic operations to augment bone healing. Autologous bone graft is the gold standard for osteogenesis, but the amount available from the patient’s iliac crest is often insufficient to fill the defect and donor site morbidity is a significant complication. Alternatively, allograft can be implanted into patients, however, processing is necessary to reduce the immunicity of the graft and the risk of transmission of infection, but this destroys osteoprogenitor cells and hence reduces the osteogenic properties of the graft. Mesenchymal stem cells (MSCs) are present in bone marrow and have the ability to differentiate into osteoblasts. Therefore our study examined the use of MCSs, from bone marrow, to enhance the osteogenic properties of allograft. Hypothesis: MSCs cultured on freeze-dried ethylene oxide treated bone allograft differentiate into osteoblasts, thereby increasing the osteogenic nature of the graft material. Method: After informed consent, bone marrow aspirates were taken from 10 patients during elective orthopaedic operations. MSCs were characterized using Stro-1 antibody and grown on freeze-dried ethylene oxide treated bone allograft in vitro. The hypothesis was tested on three groups of graft, with eight samples in each group. Firstly, freeze-dried ethylene oxide treated bone graft was tested (group 2). For a negative control, allograft was heated to 70°C to denature the osteogenic proteins (group 1). The final group tested the effect of additional osteogenic supplements (100nM dexamethasone, 0.05mM ascorbic acid and 10mM (-glycerol phosphate) on MSCs on allograft (group 3). Osteoblastic differentiation of MSCs was observed under scanning (SEM) and transmission (TEM) electron microscopy, and by measuring protein levels: alkaline phosphatase (ALP), osteopontin and type I pro-collagen over 14 days. Results: SEM confirmed that MSCs could be successfully cultured on bone allograft. Cells grown in groups 2 and 3 were characteristic of metabolically active osteoblasts and collagen extracellular matrix was observed under TEM. The amount of ALP protein produced by MSCs cultured in groups 2 and 3 increased significantly over 14 days (P< 0.05), but there was no increase in group 1. ALP, osteopontin and type I pro-collagen production was significantly greater for group 2 than for group 1 and for group 3 than for group 2 (P< 0.05). Discussion and Conclusions: ALP, type I pro-collagen and osteopontin proteins are known to be produced by osteoblasts during increasing cell maturation and the levels of each of these proteins increased significantly when MSCs were cultured on allograft for 14 days compared with the negative control. The addition of osteogenic supplements significantly increased production of these proteins. Furthermore, MSCs cultured in groups 2 and 3 produced extracellular collagen matrix. These results are consistent with allograft causing MSCs to differentiate into osteoblasts and that this differentiation increases with additional osteogenic supplements. This study confirms that MSCs, derived from autologous bone marrow, could be used to increase the osteogenic potential of allograft, thereby increasing bony healing in patients

Revision hip surgery is becoming increasingly common, 300 procedures being performed in 2001 at our institution. In order to achieve a good outcome bone stock needs to be of good quantity frequently necessitating the use of impaction bone grafting using allograft bone. Donor bone may frequently take three months before it becomes available for use due to the stringent screening procedure. Donor patients must have a clean bill of health, swabs taken at the time of surgery must obviously demonstrate no growth and blood samples taken at donation and an interval of three months, free from viral infectious diseases. It is thus easy to see the lag from the time of donation to availability and why, with increasing demand, need for allograft bone is rapidly exceeding supply. We need to look for an alternative supply of human bone allograft. We have compared the harvest of bone at the time of primary total knee replacement with that of the femoral head by both mass and volume. Sixty consecutive patients undergoing primary hip or knee arthroplasty were included in the study, and the masses and volume of the femoral heads compared with that of the total bone cuts in knee arthroplasty. The type of knee replacement used was documented as was whether the femoral head had had a bone block removed. It was found that the mass of femoral heads was 81g, that of knee cuts 95g this is a statistically significant difference; the volume of femoral heads 66ml and that of knee cuts 75ml. The volumes of bone available from knee arthroplasty cuts are at least comparable femoral heads obtained using hip replacement and could, perhaps, provide a realistic source of bone allograft

Introduction: We conducted a retrospective study at our institution to see what effect, if any, the use of impacted morsellised bone allograft technique had on the incidence of early and late infection in revision hip arthroplasty where contemporary measures were taken. Patients and Methods: This study included 120 patients. Patients were 36 male and 84 females with the mean age at the time of revision surgery was 71.4 years (range 42 – 89 SD 9.7). In all the patients their indication for revision surgery was aseptic loosening. All the patients had impacted morsellised bone allograft as part of the reconstruction used with cemented prostheses. Clinical and radiological assessments of all patients were conducted for average of four years follow up. Results: At mean follow up period of 4 years the early infection rate was 0.8% and late infection rate was 0%. Conclusion: In our study the use of morsellised bone allograft does not appear to have added risk effect on the incidence of early or late hip joint infection provided contemporary measures are taken

We retrospectively reviewed 101 consecutive patients with 114 femoral tumours treated by massive bone allograft at our institution between 1986 and 2005. There were 49 females and 52 males with a mean age of 20 years (4 to 74). At a median follow-up of 9.3 years (2 to 19.8), 36 reconstructions (31.5%) had failed. The allograft itself failed in 27 reconstructions (24%). Mechanical complications such as delayed union, fracture and failure of fixation were studied. The most adverse factor on the outcome was the use of intramedullary nails, followed by post-operative chemotherapy, resection length > 17 cm and age > 18 years at the time of intervention. The simultaneous use of a vascularised fibular graft to protect the allograft from mechanical complications improved the outcome, but the use of intramedullary cementing was not as successful. In order to improve the strength of the reconstruction and to advance the biology of host–graft integration, we suggest avoiding the use of intramedullary nails and titanium plates, but instead using stainless steel plates, as these gave better results. The use of a supplementary vascularised fibular graft should be strongly considered in adult patients with resection > 17 cm and in those who require post-operative chemotherapy

Purpose of the study: The mechanical and radiological course of bone allografts is often favorable but osteointegration properties could be improved. We associated a safe allograft with mesenchymatous bone marrow stem cells (MSC) with known osteogenic potential. The purpose of this preliminary study was to study the biocompatibility of the treated allograft, assess the osteoblastic differentiation properties of the MSC, and determine the optimal period for colonizing the bone matrix. Material and methods: The support was a safe bone allograft preserved in a collagenic grid (Osteopure™). MSC harvested by adherence were seeded in a medium favoring osteoblastic differentiation by comparison with standard culture medium. Culture conditions varied to study the influence of the presence or not of support, the culture time, or the presence of human serum in the culture medium. For each culture medium, we noted: the number of cells, osteoblastic differentiation using markers: alkaline phosphate and osteocalcin. A histological study was also performed. Results: Peak cell amplification was achieved at three weeks culture. Presence of osteoblastic differentiation markers was clearly identified in cultures grown in the presence of support material. Microscopy demonstrated that cells stimulated by the differentiation medium adhered strongly to the bone network. Histology revealed the presence of osteoblastic activity in differentiation medium with cells taking on the classical cytological aspect of osteoblasts. Cell proliferation was at least equivalent in medium with human serum as with fetal calf serum. Discussion: This study demonstrated that the allogenic matrix does not modify the capacity of human MSC for colonization and differentiation. The cell organization is optimal compared with the absence of supporting material. Use of the patient’s own serum in the culture medium was validated enabling an autologous procedure. Use of a complex cell graft appears to be optimal after three weeks of culture. This first step proves the feasibility of the concept designed to optimize the support with the patient’s own MSC. The next step is to develop an in vivo model

We analysed the cellular immune response in ten transplantations of different massive bone allografts, of which five had a poor clinical outcome. Cytotoxic T lymphocytes (CTL) and T helper lymphocytes (TH) against mismatched donor antigens were found in all patients. More importantly, CTL with a high affinity for donor antigens were found in five cases. High-affinity CTL need no CD8 molecule to stabilise the antigen binding and are strongly associated with rejection of heart and corneal transplants. Even after removal of most of the bone-marrow cells, we found high-affinity CTL and high TH frequencies. This T-cell response could be detected over a period of years. We conclude that frozen bone allografts can induce high-affinity donor-specific CTL. The present assay allows qualification and quantification of the levels of CTL and TH in the blood. This approach may be helpful in studying the effect of the immune response on the outcome of the graft

Femoral impaction bone allografting has been developed as a means of restoring bone stock in revision total hip replacement. We report the results of 75 consecutive patients (75 hips) with a mean age of 68 years (35 to 87) who underwent impaction grafting using the Exeter collarless, polished, tapered femoral stem between 1992 and 1998. The mean follow-up period was 10.5 years (6.3 to 14.1). The median pre-operative bone defect score was 3 (interquartile range (IQR) 2 to 3) using the Endo-Klinik classification. The median subsidence at one year post-operatively was 2 mm (IQR 1 to 3). At the final review the median Harris hip score was 80.6 (IQR 67.6 to 88.9) and the median subsidence 2 mm (IQR 1 to 4). Incorporation of the allograft into trabecular bone and secondary remodelling were noted radiologically at the final follow-up in 87% (393 of 452 zones) and 40% (181 of 452 zones), respectively. Subsidence of the Exeter stem correlated with the pre-operative Endo-Klinik bone loss score (p = 0.037). The degree of subsidence at one year had a strong association with long-term subsidence (p < 0.001). There was a significant correlation between previous revision surgery and a poor Harris Hip score (p = 0.028), and those who had undergone previous revision surgery for infection had a higher risk of complications (p = 0.048). Survivorship at 10.5 years with any further femoral operation as the end-point was 92% (95% confidence interval 82 to 97)

INTRODUCTION. Allograft reconstruction after resection of primary bone sarcomas has a non-union rate of approximately 20%. Achieving a wide surface area of contact between host and allograft bone is one of the most important factors to help reduce the non-union rate. We developed a novel technique of haptic robot-assisted surgery to reconstruct bone defects left after primary bone sarcoma resection with structural allograft. METHODS. Using a sawbone distal femur joint-sparing hemimetaphyseal resection/reconstruction model, an identical bone defect was created in six sawbone distal femur specimens. A tumor-fellowship trained orthopedic surgeon reconstructed the defect using a simulated sawbone allograft femur. First, a standard, ‘all-manual’ technique was used to cut and prepare the allograft to best fit the defect. Then, using an identical sawbone copy of the allograft, the novel haptic-robot technique was used to prepare the allograft to best fit the defect. All specimens were scanned via CT. Using a separately validated technique, the surface area of contact between host and allograft was measured for both (1) the all-manual reconstruction and (2) the robot-assisted reconstruction. All contact surface areas were normalized by dividing absolute contact area by the available surface area on the exposed cut surface of host bone. RESULTS. The mean area of contact between host and allograft bone was 24% (of the available host surface area) for the all-manual group and 76% for the haptic robot-assisted group (p=0.004). CONCLUSIONS. This is the first report to our knowledge of using haptic robot technology to assist in structural bone allograft reconstruction of defects left after primary bone tumor resection. The findings strongly indicate that this technology has the potential to be of substantial clinical benefit. Further studies are warranted

We report the contamination rate in the Cambridge bone bank of 35 consecutive allograft specimens, all harvested in a clean-air environment, using a strict aseptic technique and antibiotic cover. Five of 27 femoral heads taken from living donors and three of eight massive allografts taken from cadavers were found to be contaminated. The contaminated femoral heads were discarded. All massive allografts were rendered sterile by gamma-irradiation. It is important to exclude bacteriological contamination of harvested and banked bone.

Aim: Failed primary hip arthroplasty often results in significant loss of host bone. Revision surgery may require bone grafting to restore bone stock prior to insertion of a new cup. A two to five year follow up of one method of acetabular revision for severe bone stock loss is presented

Materials and Methods: Seventeen patients had acetabular revision with the use of impacted morcellised bone and a cage reconstruction with a cemented cup. The average age at the time of revision was 62. All patients were followed prospectively with regular X-rays. A variety of cages were employed and bone graft was hand morcellised from femoral heads or cadaver distal femurs.

Glenoid replacement is technically challenging. Removal of a cemented glenoid component often results in a large osseous defect which makes the immediate introduction of a revision prosthesis almost impossible. We describe a two-stage revision procedure using a reversed shoulder prosthesis. Freeze-dried allograft with platelet-derived growth factor was used to fill the glenoid defect. Radiological incorporation of the allograft was seen and its consistency allowed the placement of a screwed glenoid component. There were no signs of new mature bone formation on histological examination.

The addition of platelet-derived growth factor to the allograft seems to contribute to an increase in incorporation and hardness, but does not promote the growth of new bone.

Aim: The biological activity of demineralised bone matrix (DBM) led to the discovery of bone morphogenetic proteins (BMP). OP-1 (BMP 7) is an osteoinductive protein and has been demonstrated to be capable of inducing new bone formation in rat subcutaneous tissue and in both orthotopic and heterotopic sites in primates. In this study we have investigated whether demineralisation and addition of osteogenic. protein 1 (OP-1) improves osteoinductive properties of allograft. Methods: A randomised controlled blind trial was performed in 16 rats. One group received two pellets of fresh frozen allograft; the other received two pellets of demineralised bone (DBM) intramuscularly. In each rat one pellet was treated with OP-1 (2mg/25mg of graft). The rats were sacriþced at 28 days and tissue þxed and processed for sectioning with haematoxylin and eosin for morphology and Alcian blue and Sirrus red for collagen types I, II. Qualitative observations were made and each specimen graded 0–5 on the degree of new bone formation and integration by two blind observers. Results & Conclusions: DBM with OP-1 yielded optimal results, being signiþcantly superior to allograft alone and allograft with OP-1. DBM alone was shown to be more effective compared to the allograft preparations. Hence we have shown that demineralization and OP-1 signiþcantly improve the osteoinductive properties of allograft

This study assessed factors responsible for exclusion of patients from bone donation at primary hip arthroplasty in order to improve bone banking.

Fifty-five patients underwent screening in preoperative clinics assessing their suitability for femoral head donation. Records at the bone bank were then reviewed post operatively to check whether bone had been harvested from these individuals during surgery.

Overall, 95% of the patients screened did not proceed to bone banking. After the initial screening stage 60% of patients were excluded. The majority of exclusions (70%) were unacceptable as donors because of their potential risk of transmission of disease to recipients. Although 40% were consented for donation, femoral heads from only 5% were harvested and sent for storage in the bone bank during hip arthroplasty.

Orthopaedic surgeons must take an active part in bone banking and alternative sources of bone grafts require exploration in the future to meet the increasing demand.

Bone grafts are frequently used to augment bone healing. Autologous bone graft is the gold standard for osteogenesis but is limited by availability and donor site morbidity. The processing required to lower the immunogenicity of allograft also reduces the osteogeneic properties. Bone marrow contains mesenchymal stem cells (MSCs) which differentiate into osteoblasts, forming bone. Our study examined the use of bone marrow to enhance the osteogenic properties of allograft.

Bioactive proteins within allogenic bone graft stimulate marrow-derived MSCs to differentiate into osteoblasts, thereby increasing the osteogenic nature of the graft.

After informed consent, bone marrow aspirates were taken from five patients during orthopaedic operations. Freeze-dried ethylene oxide treated allograft, from a number of donors, was obtained from the bone bank. MSCs isolated from each marrow aspirate were grown on eight samples of test allograft. Further allograft was heated to 70°C to denature the osteogenic proteins and MSCs from each aspirate were grown on 8 samples, as a negative control. Osteoblastic differentiation of MSCs cultured on the types of allograft was compared.

Scanning electron microscopy confirmed that MSCs covered the allograft after 14 days. Transmission electron microscopy showed that cells on the test allograft were characteristic of osteoblasts and produced collagen extracellular matrix. The levels of osteoblastic proteins, ALP, osteopontin and Type I pro-collagen, produced by cells on test allograft were significantly greater compared with heat-treated control (P< 0.005), after days 7 and 14.

Our study showed that marrow-isolated MSCs could be successfully cultured on allograft. As the levels of osteoblastic proteins increased significantly when MSCs were grown on allograft, osteogenic proteins within allograft caused MSCs to change into osteoblasts. This confirms that autologous marrow MSCs could be grown on allograft to increase its osteogenic prior to grafting, resulting in increased rate of bony healing.

Introduction: Iontophoresis is a method to introduce antibiotic molecules into allograft bone using an electrical potential; the antibiotics may then be released at therapeutic levels for extended periods of time. This is the first report of iontophoresed allograft implantation into patients. Method: A method of loading tubular sections of cortical bone was used in theatre prior to implantation. Postoperative serum, drain and allograft antibiotic assays were performed. Patients were followed-up clinically and radiologically. All patients who received a bulk segmental allograft from June 1997 were entered into the trial. Results: Since June 1997, 35 patients have received 37 allografts. Indications for allograft insertion were limb salvage for tumour (18), and poor bone stock associated with infection (11), periprosthetic fracture (6), aseptic loosening (1) and recurrent dislocation of total hip replacement (1). Mean follow-up is 3.3 years, and no patients have been lost to follow-up. One patient received two allografts in different sites and one had an allograft exchange. There has been one superficial wound infection and one deep infection. The latter patient was revised to another iontophoresed allograft and has had no recurrence at 34 months. One allograft has been revised to a vascularised fibular graft and allograft exchange following fracture of metal fixation. There was one case of persistent non-union in a knee arthrodesis which was treated after 21 months by removal of the intramedullary fixation and use of an Illizarov frame. The allograft was not revised. All other allografts are in situ with no complications related to the allograft. Eleven patients had pre-existing proven infections. None of these patients have been re-infected to date. Therapeutic gentamicin and flucloxacillin levels were detected in drain fluid samples post-operatively. Conclusions: Iontophoresis is a safe and inexpensive technique that delivers high local dose of antibiotic, which may reduce infection in avascular allograft bone

Introduction and Aims: Iontophoresis is a method to introduce antibiotic molecules into allograft bone using an electrical potential. In-vitro testing has shown that these antibiotics should be released in their bioactive form at therapeutic levels for extended periods of time. This is the first report of iontophoresed allograft implantation into patients. Method: A method of loading tubular sections of cortical bone was used in theatre prior to implantation. The bone was held vertically in an antibiotic bath with a cylindrical outer electrode and a wire electrode down the centre of the bone. An electrical potential of approximately 90V was applied to drive the antibiotics into the bone. Post-operative serum, drain and allograft antibiotic assays were performed. All patients were followed-up clinically and radiologically. Patients who required a bulk segmental allograft from June 1997 to present were entered into the trial and received iontophoresed bone. Results: Since June 1997, 35 patients have received 37 iontophoresed allografts. Indications for allograft insertion were limb salvage for tumor (16), poor bone stock associated with infection (12), periprosthetic fracture (seven), aseptic loosening (one) and recurrent dislocation of total hip replacement (one). One patient had acute complications requiring amputation. No patients were lost to follow-up with a mean follow-up of 3.3 years. Two patients required an allograft exchange for fracture and infection. There were two late allograft infections at 10 and 18 months. One patient was revised to another iontophoresed allograft and has had no recurrence at two years. The other infection required above knee amputation. One allograft was revised with allograft exchange and vascularised fibular graft following fracture of metal fixation. There was one case of persistent non-union in a knee fusion, which was treated after 21 months by removal of the intermedullary fixation and allograft and use of an Ilizarov frame. All other allografts are in-situ with no complications related to the allograft. Twelve patients had pre-existing proven infections. None of these patients have been re-infected to date. Therapeutic gentamicin and flucloxacillin levels were detected in drain fluid samples post-operatively, averaging from 39.9 mg/L at two hours to 5.97mg/L at 48 hours for gentamicin and 16.83mg/L at two hours and 2.23 mg/L at 48 hours for flucloxacillin. This was significantly greater than the minimum inhibitory concentration (MIC) against Staphylococcus aureus for gentamicin (0.25mg/L) and flucloxacillin (0.30mg/L). At the same time, blood levels remained in a safe range. Conclusion: Iontophoresis is a safe and inexpensive method that can be executed in the operating theatre. Iontophoresed bone delivers a high local dose of antibiotic, which may prevent early biofilm formation initiated during allograft handling and exposure to theatre air. With no early infections and no re-infections, further assessment of this technique continues with guarded optimism

Reconstruction acetabular surgery with bone stock loss is still a difficult and challenging problem for the orthopaedic surgeon. The goals of acetabular revision are: stable bone coverage that can support the new acetabular component, restoration of the anatomy and bone stock for future revisions, equalization of leg length and restoration of the centre of hip motion. These goals are difficult to achieve when the pelvic defect is particularly severe. We examine the case of a female 73 years old who underwent a third revision arthroplasty of the hip joint because of extensive bony defect of the acetabular cavity (massive protrusio defect-type III –D’Antonio- combined segmental/cavitary acetabular defect). The femoral component which was revised in a previous operation with a mega stem (type Kotz), was radiologically stable and symptomless. Preoperative radiological assessment was performed using standard radiographic views, Judet views and CT scan. The surgical approach that we used was a slight modification of the previous incision achieving a better visualization of the entire acetabulum and iliac wing. The loose acetabular cup as well as soft tissue and debris were removed from the acetabulum. The large acetabular defect was filled with a massive allograft (tibial plateau) properly cut and shaped. The stability of the allograft was achieved fixing the allograft to the iliac bone with screws. A large amount of particulate allograft bone was placed in the depths of the acetabular defect restoring a proper level of the acetabular floor. Then a Burke-Schneider cage was firmly seated and fixed with screws in the prepared acetabular bed. A polyethylene cup was cemented into the acetabular shell. The superior part of the Kotz femoral prosthesis was also revised with a new one. Postoperatively we din not have any complications, the graft incorporation was successful with a satisfactory functional result. We believe that the use of structural allograft bone is essential for the reconstruction of large segmentalace-tabular defects. The results however are less predictable because of important technical difficulties and sometimes serious complications occur

The Scottish National Blood Transfusion Service is the main provider of bone for grafting in Scotland. Bone is procured only from live donors, following very strict selection criteria, and we have investigated whether the amount being collected was adequate.

Our current harvest of approximately 1700 femoral heads per year is shown not to be enough to meet the future demand for revision surgery of the hip. Many more of these operations are being undertaken, and impaction grafting is being used increasingly.

We have calculated the predicted rates of collection and usage for the next four to five years so that we can expand our service in a controlled fashion.

We studied the calcium content and mechanical strength of cortical bone from rats and dogs after different periods of demineralisation, showing that the rate of demineralisation differed considerably between the species. Specimens from the rat were further treated by chemical extraction and autolysis and tested for osteoinductive properties. We showed that partially demineralised cortical bone retained adequate mechanical strength, while retaining the biological effects of completely demineralised bone. This shows that it is possible to prepare allografts which have adequate mechanical strength and still retain osteo-inductive properties.

Introduction:

This study presents a series of 64 patients undergoing tibio-talo-calcaneal (TTC) fusions with a hindfoot nail to compare the times to union and complications comparing use of allograft with no allograft.

Bone morphogenic proteins (BMPs) are members of the transforming growth factor beta (TGF-beta) family and play a central role in bone formation. These morpho-gens are known to be present in bone matrix however the characteristics of their release during the grafting process has not previously been defined. The aim of this study was to determine the release BMP-7 (osteogenic protein; OP-1) from cancellous allograft that occurs during impaction grafting for revision hip arthroplasty. Forty, 10mm cubes of cancellous bone were accurately cut from the central region of 7 fresh frozen femoral heads. The cubes were centrifuged and washed to remove the marrow contents. The cubes were then individually washed and the fluid assayed for BMP-7 activity using a commercially available enzyme linked immuno-sorbent assay kit (Raybiotech Inc.). The cubes were then divided into 4 groups with samples from each femoral head in each group. Each group was subjected to strain of either 20%, 40%, 60% or 80% using a material testing machine. The cubes were then individually washed again and the wash fluid analysed for BMP-7 activity. BMP-7 activity was found to be present in all groups. Release of BMP-7 was found to increase with increasing strain. At 80% strain the mean concentration of BMP-7 released (830 pg/g) was 58% greater than that released at 60% strain (527 pg/g), 150% greater than the concentration at 40% strain (333 pg/g) and 476% greater than at 20% strain (144 pg/g). The differences between release at 80% and 40% strain and between 80% and 20% strain were statistically significant (p=0.036, p=0.002). Activity of BMP-7 in fresh frozen cancellous allograft bone has not previously been demonstrated. This study shows that the freezing and storage of femoral heads allows some maintenance of biological activity. Furthermore we have shown that BMP-7 may be released in proportion to the strain applied to the bone. This confirms that the process of impaction of bone morsels during revision hip arthroplasty may release BMPs that could aid in the incorporation and remodelling of the allograft

Introduction: Filling of bone defects is a significant challenge in Orthopaedic Surgery. Human fresh-frozen allograft is still the most effective bone graft substitution material («gold standard»), guaranteeing all essential biological and physiochemical demands (osteogenic, osteoinductive, and osteoconductive) when the necessary amount of autologous bone is not available. Using donor screening recommendations, more than 50 % of potential donors have to be excluded. With increasing incidence for revision hip surgery and especially acetabular reconstructions, a hospital associated bone bank has difficulties meeting demand. The aim of this study is to evaluate the balance and resource utilisation of a hospital associated bone bank for fresh-frozen allografts and the correlation to commercial alternatives regarding cost effectiveness.

Method: For evaluation of resource utilisation and cost effectiveness of a hospital associated bone bank, all donation processes and the details of allograft use were analysed and summarized within a period of 30 months. Given the increasing disproportion of demand and availability, the reasons for exclusion, especially for exclusion during the preservation period, were carefully scrutinized. The costs of installation and maintenance of the bone bank, as well as all costs in the screening process were balanced to calculate the «per head»-price. The results were compared to commercial alternatives.

Results: Within the period of evaluation 632 femoral heads were available for donation. Through the screening process 359 femoral heads (56.8%) met at least one criterion for exclusion. At the end of the observation period of six months and after HIV retesting, 246 allografts met all criteria for use. The mean period between inclusion in the bone bank and release was 10.9 5.0 months (range 6.0–30.8).

50.8% of released allografts (125 heads) were used in revision arthroplasty. In spine surgery 83 allografts (33.7%) were implanted in spinal fusions and for cage filling during vertebral body replacement. Thirty-two grafts (13.0%) were used in miscellaneous surgeries with minor bone demand.

The costs per donation were 92, with personnel costs the price per head was 140. The price range for commercial alternatives starts at 100 for 1 cm.

Conclusion: A hospital associated bone bank for fresh-frozen allografts is still an effective and cost effective method to maintain material for bone defect filling. To meet demand, information and communication to donors has to be increased to get the HIV-retests. Additionally, division of donations into smaller portions helps to decrease waste in surgeries where less bone is required.

Aim: This study wants to investigate whether the administration of stromal stem cells (SSC) in a platelet-rich plasma (PRP) scaffold could promote angiogenesis which resulted in a better allograft integration.

Methods: surgery: A monolateral resection of 3cm segment of the metatarsus, was perfomed in 10 adult cross-breed sheep (3–4 years old), weighting 60–70 kg.

Isolation and ex-vivo expansion of SSC: nucleated cells were isolated with density gradient and expanded ex-vivo with alpha-MEM containing 20% FCS.

Radiographic and histomorphometric analysis: Radiographs were made after surgery and after 1, 2 and 4 months. Histomorphometric studies were carried out to study the defect and the new bone formation at the implant site

Results: Union had occurred in all the 5 animals of the SSC group after 4 months as observed radiographically and morphologically, while in the control group the osteotomy line was still visible. Histomorphometric analysis demonstrated a higher % of new-bone formation in both the host (%section quadrant) and the grafted bone in SSC animals.

Conclusions: Results presented suggest that SSC in PRP-based scaffold have improved allograft integration. In conclusion the application of this surgical approach may result in an increased and accelerated bone graft integration, reducing the time required for bone healing and increasing the chances of a successful bone implant.

Despite the widespread use of demineralized bone matrix (DBM) allografts there are few clinical studies comparing DBM to iliac crest bone grafting (ICBG). A comparison of DBM to ICBG is presented in patients who underwent four corner fusions of the wrist by one surgeon using identical operating technique.

The senior author’s first fourteen consecutive patients in which DBM was used for four corner fusion were compared with fourteen patients selected from a total of 48 patients in which ICBG was used. The ICBG group was matched for age, indication and healing impairing co-morbidities (mainly smoking). Patient radiographs from the 8th, 12th and 24th postoperative week follow up were digitized and blinded. Three orthopaedic surgeons, not involved in the patients care, rated the degree of bony union in a scale of 0 (no evidence of healing) to 3 (solid bony healing). The operating technique and fixation was identical in all patients. K-wires were removed at a mean of 8.2 weeks for DBM and 7.7 weeks for the ICBG group.

All patients had a minimum follow-up of one year. All fusions healed both radiographically and clinically without complications. Review of the radiographs revealed significantly less visible healing at 8 weeks in the DBM group (mean score 1.50 versus 1.74 of the ICBG group, p< .05). Lower scores were also obtained for the DBM group at 12 and 24 weeks but they did not reach statistical significance.

In this study both DBM and ICBG were equally effective in achieving solid bone union for intercarpal fusions. However, the statistical power of this series is not adequate to conclude that healing rates are equal between the two graft materials. The radiographic appearance of bridging bone lagged behind in the DBM group. The biological significance of this finding is not clear; it could indicate delayed mineralization at the fusion site. Such a delay may be significant in graft choice for patients with healing impairment.

Periosteal mesenchymal stem cells (PMSC) are an emerging niche of stem cells to enhance bone healing by tissue engineering process. They have to be differentiated into osteoprogenitors in order to synthesize new bone matrix. In vitro differentiation with specific differentiation medium (DM) is not exactly representative of what occurs in vivo. The interaction between PMSC and growth factors (GF) present in biological matrix is somewhat less understood. The goal of this study is to explore the possibility of spontaneous PMSC differentiation in contact with different biological matrices without DM. 500.000 porcine PMSC were seeded on 6-well plates and cultured with proliferation medium (PM). When reaching 80% confluence, biological samples (n=3) of demineralized bone matrix (DBM), decellularized porcine bone allograft (AOp), human bone allograft (AOh), human periosteum (HP) and human fascia lata (HFL) were added. Negative and positive control wells included cells with only PM or DM, respectively. The differentiation progress was assessed by Alizarin Red staining at days 7, 14 and 21. Bone morphogenetic protein content (BMP 2, 4, 5, 6, 7, 8, 9 and 11) of each sample was also investigated by western blot. Alizarin red highlighted bone nodules neoformation on wells containing AOp, AOh and DBM, like positive controls. HP and HFL wells did not show any nodules. These results are correlated to a global higher BMP expression profile in AOp than in HP and HFL but not statistically significant (p=0.38 and p>.99, respectively). The highest expression in each tissue was that of BMP2 and BMP7, which play an important role in osteoinduction. PMSC are well known to participate to bone formation but, despite BMP presence in HP and HFL, they did not permit to achieve osteogenesis alone. The bone contact seems to be essential to induce in vitro differentiation into osteoprogenitors