Maintenance of vertebral mechanical stability is of paramount importance to prevent pathologic fractures and resultant neurologic compromise in individuals with spinal metastases. Current non-surgical treatments for vertebral metastases (i.e. chemotherapy, bisphophonates (BP) and radiation) yield variable responses in the tumour and surrounding bone. Photodynamic therapy (PDT) is a novel, minimally-invasive technology that utilizes a drug activated by light at a specific non-thermal wavelength to locally destroy tumour cells. Previously, we observed that PDT can ablate cancer cells within bone and yield short-term (1-week) improvements in vertebral architecture and biomechanical strength, particularly when combined with BP therapy. This study aims to evaluate the effects of PDT in vertebral bone over a longer (6-week) time period, alone and combined with previous BP treatment, to determine if improvements in skeletal architecture and strength are maintained. Fourty healthy rnu/rnu rats were randomly assigned to four treatment groups: (i) untreated control, (ii) BP only, (iii) PDT only and (iv) PDT following BP. BP treatments were administered on day 0 via subcutaneous injection of zoledronic acid. PDT was administered on day 7 via an intravenous injection of BPD-MA photosensitizer. A flat-cut optical fiber was inserted percutaneously adjacent to lumbar vertebra L2. After a 15-minute drug-light interval, 75J of light energy was delivered from a 690nm laser. Six weeks later, animals were euthanized. Structural properties of excised L2 vertebral bodies were quantified through semi-automated analysis of micro-CT images. In of the specimens, mechanical properties were evaluated by loading the L2 vertebral body to failure in axial compression. The remaining L2 vertebrae were analyzed for morphology, osteoid formation and osteoclast activity using histological methods.Purpose
Method
To develop a low complexity highly-automated multimodal approach to segment vertebral structure and quantify mixed osteolytic/osteoblastic metastases in the rat spine using a combination of CT and MR imaging. We hypothesize that semi-automated multimodal analysis applied to 3D CT and MRI reconstructions will yield accurate and repeatable quantification of whole vertebrae affected by mixed metastases. Mixed spinal metastases were developed via intra-cardiac injection of canine Ace-1 luciferase transfected prostate cancer cells in a 3 week old rnu/rnu rat. Two sequential MR images of the L1-L3 vertebral motion segments were acquired using a 1H quadrature customized birdcage coil at 60 m isotropic voxel size followed by CT imaging at a 14m isotropic voxel size. The first MR image was T1 weighted to highlight the trabecular structure to ensure accurate registration with the CT image. The second MR image was T2 weighted to optimize differentiation between bone marrow and osteolytic tumour tissue. Samples were then processed for undecalcified histology and stained with Goldners Trichrome to identify mineralized bone and unmineralized new bone formation. All images were resampled to 34.9 m and manually aligned to a global axis. This was followed by an affine registration using a Quasi Newton optimizer and a Normalized Mutual Information metric to ensure accurate registration. The whole individual vertebrae and their trabecular centrums were then segmented from the CT images using an extended version of a previously developed atlas based registration algorithm. An intensity-based thresholding method was used to segment the regions corresponding to osteoblastic tumor predominantly attached to the outside of the cortical shell. The whole vertebral segmentation from the CT was warped around the T2 weighted MR to define the bone boundaries. An intensity-based thresholding approach was then applied to the T2 weighted MR segment the osteolytic tumor.Purpose
Method
control, no treatment; BP only; PDT only; BP and PDT combined. Seven days after MT-1 injection 60 μg/kg of zoledronic acid was injected. PDT treatment was administered on day 14 using the photosensitizer BPD-MA (1.0 mg/kg; Visudyne). Fifteen minutes later, laser-light (690nm; 75J) was administered to the lumbar vertebrae. The rats were euthanized 7 days after PDT treatment. A total of 45 vertebrae were evaluated using a histomorphometric program (GENIE™, Aperio) to assess tumour burden. Statistical analyses were performed using a one-way ANOVA with a Tukey post hoc test. A p-value p<
.05 was considered to be statistically significant..
