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

Objectives. Despite promising results have shown by osteogenic cell-based demineralized bone matrix composites, they need to be optimized for grafts that act as structural frameworks in load-bearing defects. The aims of this study is attempt to assess the effects of laser perforations on osteoinduction in cortical bone allografts. Methods. Sixteen wistar rats were divided into two groups according to the type of structural bone allograft; the first: partially demineralized only (Donly) and the second: partially demineralized laser-perforated (DLP). Trans-cortical holes were achieved by Er:YAG laser at a wave length of 2.94 µm in four rows of three holes approximated cylindrical holes 0.5 mm in diameter, with centres 2.5 mm apart. Histologic and histomorphometric analysis were performed at 12 weeks. Results. Statistical analysis showed significant differences between the 2 groups at 3 months. Results showed that partially demineralized laser-perforated grafts had substantially higher incorporation by woven bone than partially demineralized grafts; yet this difference at the interface gap remained insignificant. In DLP allografts healing at the junction was more complete and a wider area was in contact with host and graft surfaces. Conclusions. Based on the results of this study, it may be concluded that surface changes induced by Er:YAG laser, accelerated bone healing with good osteoinduction results and the process might have improved, if they could have been supplemented with the proper stipulations

Background. Despite promising results have shown by osteogenic cell-based demineralized bone matrix composites, they need to be optimized for grafts that act as structural frameworks in load-bearing defects. The purpose of this experiment is to determine the effect of bone marrow mesenchymal stem cells seeding on partially demineralized laser-perforated structural allografts that have been implanted in critical femoral defects. Materials and Methods. Thirty-two wistar rats were divided into four groups according to the type of structural bone allograft; the first: partially demineralized only (Donly), the second: partially demineralized stem cell seeded (DST), the third: partially demineralized laser-perforated (DLP), and the fourth: partially demineralized laser-perforated and stem cell seeded (DLPST). Trans-cortical holes were achieved in four rows of three holes approximated cylindrical holes 0.5 mm in diameter, with centres 2.5 mm apart. P3 MSCs were used for graft seeding. Histologic and histomorphometric analysis were performed at 12 weeks. Results. DLP grafts had the highest woven bone formation, where most parts of laser pores were completely healed by woven bone. DST and DLPST grafts surfaces had extra vessel-ingrowth-like porosities. Furthermore, in the DLPST grafts, a distinct bone formation at the interfaces was noted. Conclusion. This study indicated that surface changes induced by laser perforation, accelerated angiogenesis induction by MSCs, which resulted in endochondral bone formation at the interface. Despite non-optimal results, stem cells showed a tendency to improve osteochondrogenesis, and the process might have improved, if they could have been supplemented with the proper stipulations

Background. Structural bone allografts are an established treatment method for long-bone structural defects arising from such conditions as trauma, sarcoma, and osteolysis following total joint replacement. However, the quality of structural bone allografts is difficult to non-destructively assess prior to use. The functional lifetime of structural allografts depend on their ability to resist cyclic loading, which can lead to fracture even at stress levels well below the yield strength. Because allograft bone has limited capacity for remodeling, optimizing allograft selection for bone quality could decrease long-term fracture risk. Raman spectroscopy biomarkers can non-destructively assess the three primary components of bone (collagen, mineral, and water), and may predict the resistance of donor bone allografts to fracture from cyclic loads. The purpose of this study was to prospectively assess the ability of Raman biomarkers to predict number of cycles to fracture (“cyclic fatigue life”) of human allograft cortical bone. Methods. Twenty-one cortical bone specimens were from the mid-diaphysis of human donor bone tissue (bilateral femurs from 4 donors: 63M, 61M, 51F, 48F) obtained from the Musculoskeletal Transplant Foundation. Six Raman biomarkers were analyzed: collagen disorganization, type B carbonate substitution (a surrogate for mineral maturation), matrix mineralization, and 3 water compartments. Specimens underwent cyclic fatigue testing under fully reversed conditions at 35 and 45MPa (physiologically relevant stress levels for structural allografts). Specimens were tested to fracture or to 30 million cycles (“run-out”), simulating 15 years of moderate activity (i.e., 6000 steps per day). Multivariate regression analysis was performed using a tobit model (censored linear regression) for prediction of cyclic fatigue life. Specimens were right-censored at 30 million cycles. Results. All of the 6 biomarkers that were evaluated were independently associated with cyclic fatigue life (p < 0.05). The multivariate model explained 70% of the variance in cyclic fatigue life (R2=0.695, p<0.001,). Increasing disordered collagen (p<0.001) and loosely collagen-bound water compartments (p<0.001) were associated with decreased cyclic fatigue life. Increasing type B carbonate substitution (p<0.001), matrix mineralization (p<0.001), tightly collagen-bound water (p<0.001), and mineral-bound water (p=0.002) were associated with increased cyclic fatigue life. In the predictive model, 42% of variance in cyclic fatigue life was attributable to degree of collagen disorder, all bound water compartments accounted for 6%, and age and sex accounted for 17%. Conclusions. Raman biomarkers of three bone components (collagen, mineral, and water) predict cyclic fatigue life of human cortical bone. Increased baseline collagen disorder was associated with decreased cyclic fatigue life, and was the strongest determinant of cyclic fatigue life. Increased matrix mineralization and mineral maturation were associated with increased cyclic fatigue life. Bound-water compartments of bone contributed minimally to cyclic fatigue life. These results are complementary with prior Raman studies of monotonic testing of bone that reported decreased toughness and strength with increased collagen disorder and increased stiffness with increased bone mineralization and mineral maturation. This model should be prospectively validated. Raman analysis is a promising tool for the non-destructive evaluation of structural bone allograft quality and may be useful as a screening tool for selection of allograft bone. Acknowledgements. Supported by a grant from the Musculoskeletal Transplant Foundation. The Dudley P. Allen Fellowship (JYD), Wilbert J. Austin Professor of Engineering Chair (CMR) and the Leonard Case Jr. Professor of Engineering Chair (OA) are gratefully acknowledged

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