We undertook a study of the anti-tumour effects of hyperthermia, delivered via magnetite cationic liposomes (MCLs), on local tumours and lung metastases in a mouse model of osteosarcoma. MCLs were injected into subcutaneous osteosarcomas (LM8) and subjected to an alternating magnetic field which induced a heating effect in MCLs. A control group of mice with tumours received MCLs but were not exposed to an AMF. A further group of mice with tumours were exposed to an AMF but had not been treated with MCLs. The distribution of MCLs and local and lung metastases was evaluated histologically. The weight and volume of local tumours and the number of lung metastases were determined. Expression of heat shock protein 70 was evaluated immunohistologically. Hyperthermia using MCLs effectively heated the targeted tumour to 45°C. The mean weight of the local tumour was significantly suppressed in the hyperthermia group (p = 0.013). The mice subjected to hyperthermia had significantly fewer lung metastases than the control mice (p = 0.005). Heat shock protein 70 was expressed in tumours treated with hyperthermia, but was not found in those tumours not exposed to hyperthermia. The results demonstrate a significant effect of hyperthermia on local tumours and reduces their potential to metastasise to the lung.
Periprosthetic osteolysis has attracted attention as a cause of loosening after arthroplasty. The aim of the present study was to examine inflammatory cell localization and the occurrence of apoptosis in granulation tissue from patients who required revision arthroplasty due to loosening caused by osteolysis. 7 patients were studied comprising 3 patients who underwent FHR and 4 patients who underwent THR. Their mean age at the time of surgery was 63.6 years. The mean period from their previous operation to revision was 8.8 years. Granulation tissue was collected from around the loosened implant fixed in 4% paraformaldehyde and embedded in paraffin. Sections were cut and were first stained with hematoxylin and eosin. Next, immunohistochemical studies were performed using the avidin-biotin complex method. CD45 was used as the primary antibody to detect T cells, and CD68 was used to detect macrophage-like cells. The activity of the macrophage-like cells was assessed with anti-I-NOS and anti-MMP-9. Apoptosis was investigated using anti-single-stranded DNA (ssDNA). Using another granulation tissue was stored at −80%C, DNA was extracted, and the presence of DNA fragmentation was detected by agarose gel electrophoresis. Vascularization and infiltration by a large number of inflammatory cells were seen along with large multinucleated osteoclas-like cells. Immunohistochemical studies revealed CD45-positive cells primarily around the blood vessels. The CD68-positive cells were mainly multinucleated cells. The multinucleated cells were i-NOS-positive in 4 patients, and were MMP-9-positive in 5 patients. The nuclei of many of the multinucleated cells were positive for ssDNA. Agarose gel electrophoresis of DNA showed a marked ladder pattern at the 170 base pair region. This finding indicated DNA fragmentation or apoptosis. Apoptotic cells were seen in granulation tissue harvested from around loosened implants suggesting that apoptosis may play a role in the pathophysiology of osteolysis.
