Residual tumor cells left in the bone defect after malignant bone tumor resection can result in local tumor recurrence and high mortality. Therefore, ideal bone filling materials should not only aid bone reconstruction or regeneration, but also exert local chemotherapeutic efficacy. However, common bone substitutes used in clinics are barely studied in research for local delivery of chemotherapeutic drugs. Here, we aimed to use facile manufacturing methods to render polymethylmethacrylate (PMMA) cement and ceramic granules suitable for local delivery of cisplatin to limit bone tumor recurrence. Porosity was introduced into PMMA cement by adding 1-4% carboxymethylcellulose (CMC) containing cisplatin, and chemotherapeutic activity was rendered to two types of granules via adsorption. Then, mechanical properties, porosity, morphology, drug release kinetics, ex vivo reconstructive properties of porous PMMA and in vitro anti-cancer efficacy against osteosarcoma cells were assessed. Morphologies, molecular structures, drug release profiles and in vitro cytostatic effects of two different drug-loaded granules on the proliferation of metastatic bone tumor cells were investigated. The mechanical strengths of PMMA-based cements were sufficient for tibia reconstruction at CMC contents lower than 4% (≤3%). The concentrations of released cisplatin (12.1% and 16.6% from PMMA with 3% and 4% CMC, respectively) were sufficient for killing of osteosarcoma cells, and the fraction of dead cells increased to 91.3% within 7 days. Functionalized xenogeneic granules released 29.5% of cisplatin, but synthetic CaP granules only released 1.4% of cisplatin over 28 days. The immobilized and released cisplatin retained its anti-cancer efficacy and showed dose-dependent cytostatic effects on the viability of metastatic bone tumor cells. Bone substitutes can be rendered therapeutically active for anticancer efficacy by functionalization with cisplatin. As such, our data suggest that multi-functional PMMA-based cements and cisplatin-loaded granules represent viable treatment options for filling bone defects after bone tumor resection

We evaluated the possible induction of a systemic immune response to increase anti-tumour activity by the re-implantation of destructive tumour tissue treated by liquid nitrogen in a murine osteosarcoma (LM8) model. The tumours were randomised to treatment by excision alone or by cryotreatment after excision. Tissue from the tumour was frozen in liquid nitrogen, thawed in distilled water and then re-implanted in the same animal. In addition, some mice received an immunological response modifier of OK-432 after treatment. We measured the levels of interferon-gamma and interleukin-12 cytokines and the cytotoxicity activity of splenocytes against murine LM8 osteosarcoma cells. The number of lung and the size of abdominal metastases were also measured. Re-implantation of tumour tissue after cryotreatment activated immune responses and inhibited metastatic tumour growth. OK-432 synergistically enhanced the anti-tumour effect. Our results suggest that the treatment of malignant bone tumours by reconstruction using autografts containing tumours which have been treated by liquid nitrogen may be of clinical value

Concomitant tumour resistance (CTR) is a unique phenomenon in which animals harbouring large primary tumours are resistant to the growth of smaller metastatic tumours by systemic angiogenic suppression. To examine this clinically, in ten patients with osteosarcoma, we investigated the effects of removal of the primary tumour on the development of pulmonary metastases, the systemic angiogenesis-inducing ability and the serum levels of several angiogenesis modulators. We found that removal of the primary tumour significantly elevated systemic angiogenesis-inducing ability in five patients who had post-operative recurrence of the tumour. Post-operative elevation of the angiogenesis-induced ability was suppressed by the addition of an angiogenic inhibitor, endostatin. Also, primary removal of the tumour decreased the serum levels of vascular endothelial growth factor and endostatin. These findings suggest, for the first time, the presence of CTR in patients with osteosarcoma for whom postoperative antiangiogenic therapy may be used to prevent the post-operative progression of micrometastases

Summary. This is the first ever study to report the successful elimination of malignant cells from salvaged blood obtained during metastatic spine tumour surgery using a leucocyte depletion filter. Introduction. Catastrophic bleeding is a significant problem in metastatic spine tumour surgery (MSTS). However, intaoperative cell salvage (IOCS) has traditionally been contraindicated in tumour surgery because of the theoretical concern of promoting tumour dissemination by re-infusing tumour cells into the circulation. Although IOCS has been extensively investigated in patients undergoing surgery for gynaecological, lung, urological, gastrointestinal, and hepatobiliary cancers, to date, there is no prior report of the use of IOCS in MSTS. We conducted a prospective observational study to evaluate whether LDF can eliminate tumour cells from blood salvaged during MSTS. Patients & Methods. After Institutional Review Board (IRB) approval, 21 consecutive patients with metastatic spinal tumours from a known epithelial primary (defined as originating from breast, prostate, thyroid, renal, colorectal, lung, nasopharyngeal) who were scheduled for MSTS were recruited with informed consent. During surgery, a IOCS device (Dideco, Sorin Group, Italy) was used to collect shed blood from the operative field. Salvaged blood was then passed through a leucocyte depletion filter (RS1VAE, Pall Corporation, UK). 15-ml specimens of blood were taken from each of three consecutive stages: (i) operative field prior to cell saver processing (Stage A); (ii) transfusion bag post-cell saver processing (Stage B); (iii) filtered blood after passage through LDF (Stage C). Cell blocks were prepared by the pathology department using a standardised laboratory protocol. From each cell block, 1 haematoxylin and eosin (H&E) slide, and 3 slides each labelled with one of the following monoclonal mouse cytokeratin antibodies AE1/3, MNF 116 and CAM 5.2 were prepared. The cytokeratin antibodies are highly sensitive and specific markers to identify tumour cells of epithelial origin. These slides were read by one of two consultant pathologists who were provided full access to information on operative notes, but were blinded to the actual stages from which the slides were derived. Results. One case was excluded when the final diagnosis was revised to infection instead of metastatic spine tumour. Of the remaining cases, 7/21 tested positive for tumour cells in Stage A, 2 positive in Stage B. No specimen tested positive for tumour cells in Stage C. In 5 cases, posterior instrumentation without tumour manipulation was performed. Discussion/Conclusion. In this first-ever study of cell saver use in spine tumour surgery, we prove that leucocyte-depletion filters (LDF) can effectively eliminate tumour cells from blood salvaged during MSTS. It is now possible to conduct a clinical trial to evaluate IOCS-LDF use in MSTS. Our results are consistent with published results of similar studies performed on IOCS and LDF use outside the field of orthopaedic surgery. Spinal metastases originate from a myriad of primary cancers across various organ systems. If LDF can remove tumour cells from blood salvaged during surgery for spinal metastasis of different histological origin, then the finding can likely be extrapolated to several other fields of surgery where IOCS and LDF have not yet been attempted such as: neurosurgery, otolaryngology and general musculoskeletal oncology. Our results form a proof-of-concept for a paradigm shift in thinking regarding autotransfusion during spine tumour surgery

Introduction. Femoral neck fracture (FNF) is a common trauma in the elderly individuals. When the blood supply to the femoral head is impaired with a fracture event, the reduction or disruption of blood supply to the bone, hypoxia, leads to death of the bone marrow and trabecular bone, and eventual late segmental collapse. In the reparative process, osteoblasts and osteoclasts perform the important function of repairing the fracture site at the femoral neck. However, the reparative reaction including angiogenesis and osteogenesis remains unknown. In order to investigate the reparative reaction in patients with FNF, the distribution of tartrate resistant acid phosphatase (TRAP)-positive cells and expression of HIF-1 alpha, VEGF, and FGF-2 were observed in 36 hips in 35 patients. Methods. There were 6 men and 30 women who had a mean age of 79 years (range, 58 to 94 years). There were 10 hips with Garden stage 3, and 26 hips with Garden stage 4. The mean duration from onset to the surgery was 12 days (range: 1 to 82 days). Hematoxylin eosin staining, TRAP staining, immunohistochemistry using anti HIF-1 alpha, anti VEGF, and anti FGF-2 antibodies were performed for retrieved whole femoral heads. As a control, one femoral head in a patient who underwent wide resection for metastatic acetabular tumor was used. Results. TRAP-positive cells were detected in 9 hips (25%). They existed around the fracture site in 4 hips, while around the teres insertion and retinaculum in 7 hips. In the control, the TRAP-positive cells were detected slightly around the teres insertion and retinaculum. Around the fracture site, HIF-1 alpha expression was detected in 14 hips (39%) mainly at the vessel epithelium, VEGF was expressed at the edematous area in 28 hips (78%), while FGF-2 was detected widely in the marrow cells in 23 hips (64%). There were significant differences in TRAP staining and in HIF-1 alpha expression between Garden stage 3 and stage 4 hips, although there were no significant differences concerning the preoperative duration after fracture. Conclusion. TRAP-positive cells were recruited through the teres insertion and the retinaculum around the fracture site. HIF-1 alpha expression was detected at the restricted area around the fracture site, while VEGF and FGF-2 were detected widely regardless of Garden classification

Summary. There is emerging evidence of successful application of IOCS and leucocyte depletion filter in removing tumour cells from blood salvaged during various oncological surgeries. Research on the use of IOCS-LDF in MSTS is urgently needed. Introduction. Intra-operative cell salvage (IOCS) can reduce allogeneic blood transfusion requirements in non-tumour related spinal surgery. However, IOCS is deemed contraindicated in metastatic spine tumor surgery (MSTS) due to risk of tumour dissemination. Evidence is emerging from different surgical specialties describing the use of IOCS in cancer surgery. We wanted to investigate if IOCS is really contraindicated in MSTS. We hereby present a systematic literature review to answer the following questions: 1. Has IOCS ever been used in MSTS? 2. Is there any evidence to support the use of IOCS in other oncologic surgeries?. Methods. A systematic review of the English literature was conducted using computer searching of databases: Medline, Embase, the Cochrane Central Register of Controlled Trials for articles published between 1 January 1986 and 31 Dec 2012. Results. Question 1: A comprehensive literature search did not provide any publication describing the use of IOCS in MSTS. The application of IOCS in MSTS has never been described before. Question 2: Our systematic review shows that the use of IOCS has been extensively investigated in patients undergoing surgery for gynaecological, lung, urological, gastrointestinal, and hepatobiliary cancers. The literature review considered 281 abstracts from the initial search. After consideration by consensus, 30 articles were included in the final analysis. We included in our review -prospective, retrospective studies and in vitro studies. The selected articles were then classified according to the surgical specialty: gynaecological, lung, urological, gastrointestinal, and hepatobiliary cancers and type of studies: reinfusion studies, non-reinfusion studies and in vitro studies. 23 Reinfusion studies: Studies where salvaged blood was actually re-infused into patients and analyzed on the basis of clinical outcomes like survival, recurrence, metastasis rates, and transfusion requirements, etc. IOCS has been extensively investigated in several large cohort studies and large case series with considerable follow-up duration across urological, gynaecological, hepatobiliary and gastrointestinal cancers. Patients receiving salvaged blood have been shown to perform as well or better across a variety of clinical outcome measures as mentioned above. 2 in vitro studies and 5 non-reinfusion studies: Studies where salvaged blood was not re-infused into patients but was analyzed for the presence or viability of tumour cells in the processed blood. They consistently demonstrated the utility of LDF in either greatly reducing the number of tumour cells or even completely eradicating tumour cells from blood-tumour admixtures or salvaged blood. This provides the “proof-of-concept” that LDF is able and is effective in removing tumour cells from blood. Discussion/Conclusion. There is strong evidence that LDF can safely remove tumour cells from salvaged blood. IOCS in patients undergoing cancer surgery is not associated with any adverse clinical outcomes. The reluctance of spine surgeons to use IOCS in MSTS appears to be unsupported. There is ample evidence supporting the use of IOCS in oncological surgeries. Research is needed to evaluate the application of IOCS in MSTS

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.