Christian Delloye

Bone allografts: What they can offer and what they cannot.

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.

Pelvic reconstruction with a structural pelvic allograft after resection of a malignant bone tumor.

BACKGROUND Reconstruction of the pelvic arch after resection of a malignant pelvic tumor remains a major surgical challenge because of the high rate of associated complications. The purpose of this investigation was to assess the functional outcome and complication rate following treatment with a bone allograft to reconstruct the pelvis. METHODS Twenty-four consecutive patients underwent excision of a malignant pelvic bone tumor and reconstruction with a pelvic bone allograft. The living patients were followed for a minimum of twenty-four months. There were nineteen primary malignant bone tumors, sixteen of which were high-grade sarcomas, and there were five isolated metastases. Patients were examined clinically and radiographically and were assessed functionally with the Musculoskeletal Tumor Society score. RESULTS The mean age of the patients at the time of the index surgery was thirty-four years, and the mean duration of follow-up was forty-one months. Eighteen of the twenty-four resections involved the periacetabular area and were followed by reconstruction either with a hip prosthesis (thirteen) or with an osteochondral allograft alone (five). The six other resections involved the iliac bone. All patients received a massive bone allograft that had been sterilely procured without secondary irradiation. At the time of our last evaluation, eight patients were alive and free of disease. Seven patients had a local recurrence. Neurological deficits were present in six patients, and three had a deep infection. Nonunion of three of the sixteen allografts that could be evaluated was observed. Neither graft fracture nor lysis was observed. Eleven patients underwent surgical revision, with nine of these revisions related to the reconstruction. The average Musculoskeletal Tumor Society score at the time of the latest follow-up was 73% of the maximal possible score. The average score was 82% for the eleven patients with an age of less than twenty years at the time of the index procedure and 65% for the thirteen older patients. Ten patients walked without any assistive device, and five of them had normal function with no or only a slight limp. CONCLUSIONS Pelvic reconstruction after a limb-sparing resection is associated with a high risk of surgical complications and usually should be reserved for patients with a primary bone sarcoma. A pelvic allograft can restore the anatomy and provide good functional results, especially in young patients. Nonunion was the most common allograft-related complication.

Bone regenerate formation in cortical bone during distraction lengthening. An experimental study.

The aim of this study was to delineate the pattern of bone regeneration from cortical bone segments during distraction lengthening. The lengthening procedure was applied for various periods through the Ilizarov system on the forearms of mature dogs. Bone was sectioned either by corticotomy, preserving the nutrient artery integrity, or by osteotomy. When an osteotomy was performed, the marrow cavity was in some cases plugged with either resorbable bone wax or nonresorbable material. Under distraction, both periosteal and medullary callus on either side of the gap gave rise to new bone trabeculae. The trabeculae on either side were oriented along the direction of distraction and progressively approached one another. This striated callus emerging from both sides was the most characteristic pattern of bone regeneration subsequent to distraction lengthening. Fusion was achieved approximately four weeks after the end of the lengthening period. Most of the new bone was formed by membranous ossification; some cartilaginous nodules developed. Corticalization of the bone trabeculae that had begun at three months was not fully achieved at five months after the lengthening period. There were no differences found in the pattern of bone healing and the amount of newly formed bone after corticotomy or osteotomy with or without resorbable bone wax plugging.

Effect of sterilization on osteoinduction. Comparison of five methods in demineralized rat bone.

The aim of this study was to find a safe, effective sterilization method that does not destroy the bone-inductive capacity of demineralized bone implants. Five sterilizing agents were tested in rats. Implants procured and processed under sterile conditions served as controls. New bone formation was evaluated by determining dry weight, calcium content, and Sr-85 incorporation of the induced ossicles. Glutaraldehyde solution, formaldehyde gas, and ethylene oxide destroyed almost all the bone-inductive capacity. Irradiation by 2.5 Mrads Co-60 resulted in a loss of about half of the inductive capacity. Merthiolate (0.18 per cent) was the only sterilizing agent that did not reduce the bone-inductive capacity of the demineralized implants. Because merthiolate is not sporicidal, gamma irradiation appears to be the most appropriate sterilizing agent for demineralized bone in clinical use.

Antibiotic-loaded plaster of Paris implants coated with poly lactide-co-glycolide as a controlled release delivery system for the treatment of bone infections.

Summary. Plaster of Paris implants containing vancomycin (60 mg/g of carrier) were prepared in order to be used as local delivery system for the treatment of bone infections. The regulation of the release rate was performed by coating the carrier with a polylactide-co-glycolide polymer composed by 10% (w/w) polyglycolic acid and 90% (w/w) racemic poly (D,L-lactic acid). The release of the antibiotic from the biodegradable matrix was evaluated in vitro. From this investigation, it is clear that the drug elution depends on the coating depth. After a burst effect occurring on the first day of the experiment, therapeutic concentrations were measured during one week when uncoated implants were used. The coating allowed decrease of the burst effect and extended efficient release to more than five weeks when the implants were embedded with six layers (162 μm) of PLA45GA10. This delivery system was implanted into the femoral condyle of rabbits. It was shown that the in vivo release was also closely regulated by the coating depth. In all bone tissues (bone marrow and cortical bone) surrounding the pellets, the drug concentration exceeded the Minimum Inhibitory Concentration for the common causative organisms of bone infections (Staphylococcus aureus) for at least four weeks without inducing serum toxic levels. Due to its cheapness, facility of use and sterilization, biocompatibility and biodegradability, plaster of Paris coated with PLA45GA10 polymer giving a controlled release of vancomycin appears to be a promising sustained release delivery system of antibiotics for the treatment of bone and joint infections.Résumé. Des implants de plâtre de Paris chargés de vancomycine (60 mg/g) ont été préparés pour l’administration locale et prolongée d’antibiotique. La libération du principe actif à partir de ces vecteurs biodégradables a étéévaluée in vitro et in vivo au niveau osseux. La régulation de la diffusion de l’antibiotique a été réalisée en enrobant les implants d’épaisseurs croissantes de polymère biodégradable (PLA 45GA10) composé de 10% (p/p) d’acide glycolique et de 90% (p/p) d’un mélange racémique d’acide lactique (formes D et L). Des expériences menées in vitro, il apparaît que l’élution de la vancomycine dépend de l’épaisseur de l’enrobage de PLA45GA10. Ainsi la libération de l’antibiotique à partir des implants dépourvus d’enrobage s’effectue rapidement dès le premier jour et dure pendant une semaine. L’enrobage permet de réduire cette libération massive du début et de prolonger la libération au-delà de cinq semaines lorsque les implants sont enrobés de six couches (162 μm) de PLA45GA10. Les expériences menées in vivo, après implantation des billes de plâtre dans le condyle fémoral de lapins, confirment l’importance de l’épaisseur de la couche d’enrobage dans la régulation de la diffusion de l’antibiotique. Que ce soit au niveau de la moelle osseuse ou de l’os cortical environnant l’implant, des concentrations thérapeutiques ont été mesurées pendant plus de quatre semaines sans jamais induire de concentrations sériques toxiques. Grâce à son coût réduit, sa facilité d’utilisation et de stérilisation ainsi que son caractère biocompatible et biodégradable, le plâtre de Paris enrobé de PLA45GA10 constitue un vecteur de choix pour le traitement local et prolongé des infections osseuses.

Inhomogeneity of Human Vertebral Cancellous Bone: Systematic Density and Structure Patterns Inside the Vertebral Body

In the spine, cancellous bone quality is usually assessed for the whole vertebral body in a transverse central slice. Correct identification and assessment of the weakest parts of the cancellous bone may lead to better prediction of fracture risk. The density and structural parameters were systematically investigated inside the thoracic (T-9), thoracolumbar (T12-L1), and lumbar (L-4) vertebral bodies of nine subjects. On both sides of the median sagittal plane, anterior and posterior 8.2 mm vertical cores were harvested in the thoracic vertebra. In the thoracolumbar and lumbar vertebrae, external samples were also cored. Peripheral quantitative computed tomographic (pQCT) density analysis of the 136 cores was performed at four different levels, from the lower to the upper endplate. The relatively thin slice thickness (300 microm) and small pixel size (70 microm x 70 microm) was considered sufficient to investigate the structural parameters on the four transverse slices and in the sagittal and coronal planes (total of 816 images). Using a constant threshold a binary image was generated and the morphometric data were extracted. The binary image was further skeletonized and classical strut analysis was performed. Cancellous bone density was 20% higher in the posterior cores than in the anterior and external cores. Moreover, clear vertical inhomogeneity was noted because the lowest half of the vertebral body presented lower density than the upper half (differences ranging from 25% to 15%). All structural parameters were strongly dependent on the location of the measurement. Structural differences between anterior, posterior, and external areas were mild and followed the density patterns. On the other hand, vertical inhomogeneity of the structural parameters was important. For example, in the thoracolumbar and lumbar vertebrae, the numbers of nodes or node-to-node struts were almost twofold higher in the inferior half than in the superior half (p < 0.01), whereas trabecular thickness and number of free-ends presented a center/close-to-endplate structural pattern, with central trabeculae being 15% thicker (p < 0.05) and presenting 30% fewer free-ends (p < 0.01) than the close-to-endplate ones. Variability of density and structural parameters was high and a substantial part of this variability could be explained by the place inside the vertebral body where the measurement was made. The weak part was not in the center of the body but in its upper half where the lower density did not seem to be compensated by a higher structural architecture. Further clinical investigation could enhance fracture prediction by tracking and focusing on the weakest part of the vertebral body.

Treatment of aneurysmal bone cysts by introduction of demineralized bone and autogenous bone marrow.

BACKGROUND On the assumption that an aneurysmal bone cyst has an intrinsic potential to heal by ossification, a new, minimally invasive protocol was developed. Demineralized bone powder mixed with bone-marrow aspirate was introduced into the cyst to halt the expansion phase and to allow the cyst to ossify. We hypothesized that, in order to induce bone-healing, cells from the cyst are needed to respond to the inductive material but that curettage or extensive surgery is not necessary. The goals of the present study were to assess cyst-healing and to determine the prevalence of recurrence associated with this new procedure. METHODS Thirteen biopsy-proven primary aneurysmal bone cysts were entered through a small incision, and a paste of demineralized bone and autologous bone marrow was introduced with an applicator. The study group included three male and ten female patients with a mean age of 16.6 years. The cyst was located in a long bone in six patients, the pelvis in five patients, and the scapular glenoid and the calcaneus in one patient each. Five patients had not received treatment previously, whereas one had had a preoperative embolization and seven had recurrent lesions that had been treated previously. RESULTS After a mean duration of follow-up of 3.9 years, healing was achieved in eleven patients. CONCLUSIONS This minimally invasive method is able to promote the self-healing of a primary aneurysmal bone cyst. As no curettage is required, the proposed treatment avoids extensive surgery and blood loss and is convenient for the treatment of poorly accessible lesions such as those occurring in the pelvis. LEVEL OF EVIDENCE Therapeutic Level IV.

The enhanced performance of bone allografts using osteogenic-differentiated adipose-derived mesenchymal stem cells

Adipose tissue was only recently considered as a potential source of mesenchymal stem cells (MSCs) for bone tissue engineering. To improve the osteogenicity of acellular bone allografts, adipose MSCs (AMSCs) and bone marrow MSCs (BM-MSCs) at nondifferentiated and osteogenic-differentiated stages were investigated in vitro and in vivo. In vitro experiments demonstrated a superiority of AMSCs for proliferation (6.1±2.3 days vs. 9.0±1.9 days between each passage for BM-MSCs, respectively, P<0.001). A significantly higher T-cell depletion (revealed by mixed lymphocyte reaction, [MLR]) was found for AMSCs (vs. BM-MSCs) at both non- and differentiated stages. Although nondifferentiated AMSCs secreted a higher amount of vascular endothelial growth factor [VEGF] in vitro (between 24 and 72 h of incubation at 0.1-21% O(2)) than BM-MSCs (P<0.001), the osteogenic differentiation induced a significantly higher VEGF release by BM-MSCs at each condition (P<0.001). After implantation in the paraspinal muscles of nude rats, a significantly higher angiogenesis (histomorphometry for vessel development (P<0.005) and VEGF expression (P<0.001)) and osteogenesis (as revealed by osteocalcin expression (P<0.001) and micro-CT imagery for newly formed bone tissue (P<0.05)) were found for osteogenic-differentiated AMSCs in comparison to BM-MSCs after 30 days of implantation. Osteogenic-differentiated AMSCs are the best candidate to improve the angio-/osteogenicity of decellularized bone allografts.

Surgical inaccuracy of tumor resection and reconstruction within the pelvis: an experimental study.

Background and purpose Osseous pelvic tumors can be resected and reconstructed using massive bone allografts. Geometric accuracy of the conventional surgical procedure has not yet been documented. The aim of this experimental study was mainly to assess accuracy of tumoral resection with a 10-mm surgical margin, and also to evaluate the geometry of the host-graft reconstruction. Methods An experimental model on plastic pelvises was designed to simulate tumor resection and reconstruction. 4 experienced surgeons were asked to resect 3 different tumors and to reconstruct pelvises. 24 resections and host-graft junctions were available for evaluation. Resection margins were measured. Several methods were created to evaluate geometric properties of the host-graft junction. Results The probability of a surgeon obtaining a 10-mm surgical margin with a 5-mm tolerance above or below, was 52% (95% CI: 37–67). Maximal gap, gap volume, and mean gap between host and graft was 3.3 (SD 1.9) mm, 2.7 (SD 2.1) cm3 and 3.2 (SD 2.1) mm, respectively. Correlation between these 3 reconstruction measures and the degree of contact at the host-graft junction was poor. Interpretation 4 experienced surgeons did not manage to consistently respect a fixed surgical margin under ideal working conditions. The complex 3-dimensional architecture of the pelvis would mainly explain this inaccuracy. Solutions to this might be to increase the surgical margin or to use computer- and robotic-assisted technologies in pelvic tumor resection. Furthermore, our attempt to evaluate geometry of the pelvic reconstruction using simple parameters was not satisfactory. We believe that there is a need to define new standards of evaluation.

Massive Bone Allografts in Large Skeletal Defects After Tumor Surgery - a Clinical and Microradiographic Evaluation

SummaryMassive deep-frozen bone allografts were implanted in 13 patients after en bloc tumor resection. Patients were followed up for 14 months to 17 years. Most of the reconstructive procedures included a segmental bone allograft with knee or ankle fusion. Graft infections were the most critical complications in regard to the end results, finally requiring amputation in two cases. There were three stress fractures; two of which were successfully treated without further complication. Graft incorporation was assessed by bone scintimetry in four cases. Isotope uptake by the center of the graft was found to be superior to control bone segments at only 15 years after surgery. Two recovered allograft specimens were available for a microradiographic study. Creeping substitution was a very slow process, initiated at the outer surface of the graft and characterized at 2–3 years after implantation by large, incompletely filled osteons. The present investigation demonstrates that massive bone allografts are very slowly revascularized and are intimately anchored by the host bone. Provided that tumor control is effective and graft infection is avoided, reconstructive surgery with massive bone allografts represents a successful alternative to prosthetic implants in young adults with a long life expectancy.