Osteosarcoma is the most common malignant bone tumour in children and young people. Approximately 40% patients respond poorly to highly toxic preoperative MAP (methotrexate adriamycin, cisplatin) chemotherapy with consequent inferior survival. The role of genetic polymorphisms in drug response and toxicity is reported in acute leukaemia and some solid tumours. Recent evidence in osteosarcoma suggests increased chemotherapy toxicity is associated with improved survival. The aim of this pilot study is to investigate the influence of drug target and metabolising gene polymorphisms on tumour response and chemotherapy toxicity in osteosarcoma.

Patients who have completed MAP chemotherapy are eligible. Chemotherapy toxicity (CTCAE grade) is collected from patient records. Tumour response is graded as good (> 90% necrosis) or poor (< 90% necrosis) in resection specimen. Peripheral blood DNA is typed for genome-wide single nucleotide polymorphisms (SNP) using the Illumina 610 Quad array and analysed using Bead Studio software. Standard PCR techniques are used to genotype the Thymidylate synthase (TS) gene (folate pathway) for the presence of 2 or 3 copies of a 28 base pair repeat (2R/3R) and a G/C SNP in the 3R allele.

52 patients have entered to date: 33 good responders, 12 poor and 7 unevaluable for response. Median age 18 years (range 10–51), males:females 1.3:1. Median follow up is 39 months (range 2–76) with 11 patients relapsing. 23 patients have TS genotype 2R/2R, nineteen 2R/3R, six 3R/3R, three 2R/4R and one 2R/7R. Neither TS repeat or G/C SNP genotype correlated with histological response or degree of methotrexate stomatitis. Interestingly, presence of the 2R allele was significantly related to relapse (p=0.01) but may reflect small patient numbers. Recurrent methotrexate stomatitis (> 2 episodes of CTCAE grade 2) was weakly correlated with no relapse (p=0.07). Analysis of SNP array data with emphasis on MAP pathway polymorphisms will be presented when complete.

Introduction: The use of anterior vertebral staples in the fusionless correction of scoliosis has received increased attention in recent literature. Several animal studies have shown stapling to be effective in modulating vertebral growth. In 2005 Betz (1) published the only clinical series to date. Despite the increasing volume of literature suggesting the efficacy of this treatment, little is known about it’s biomechanical consequences. In 2007 Puttlitz (2) measured the change in spinal range of motion after staple insertion in a bovine model. They found a small but statistically significant decrease in range of motion in axial rotation and lateral bending. The clinical significance of this is questionable as the differences were only a few degrees over three vertebral levels. A well designed biomechanical evaluation of the effects of staple insertion on spinal stability is needed. The aim of this study was to evaluate the effect of insertion of a laterally placed anterior vertebral staple on the stiffness characteristics of a single motion segment.

Methods: Four-pronged shape memory alloy staples were inserted into fourteen individual bovine thoracic motion segments. A displacement controlled six degree-of-freedom robotic facility was used to test control and staple constructs through a pre-determined range of motion in flexion, extension, lateral bending, and axial rotation. All data were synchronised with robot position data and filtered using moving average methods. The stiffness in each condition was calculated in units of Nm/degree of rotation. Paired t-tests were used to compare results.

Results: Stiffness measurements in the control condition correlated with previously published measures (3). A significant decrease in stiffness (p< 0.05) following staple insertion was found in flexion, extension, lateral bending away from the staple, and axial rotation away from the staple. Stiffness for axial rotation towards the stapled side was significantly greater than for away. A near significant increase in lateral bend stiffness away from the staple compared with towards was also seen.

Discussion: These results suggest that staple insertion consistently decreased stiffness in all directions of motion. This is contrary to the results of Puttlitz (2), which reported a reduced range of motion (i.e. increased stiffness) for some motions using moment-controlled testing. This decrease in stiffness could not be explained by changes in anatomy or tissue properties between specimens, as each stapled motion segment was compared with its own intact state. Addition of the staple would intuitively be expected to increase motion segment stiffness, however we suggest that the staple prongs may cause sufficient disruption to the vertebral bodies and endplates to slightly reduce overall stiffness. Hence, growth modulation may be achieved through physical disruption of the endplate, rather than static mechanical stress. Further research is planned to investigate the proportion of load carried by the staple during spinal movement and the anatomical effect of the staple on the physis. In conclusion, anterior vertebral stapling causes a slight but significant decrease in the stiffness of treated motion segments.