Introduction. Vascularized fibular grafting following tumor resection is an essential treatment option in limb salvage surgery. This study aimed to assess the surgical and oncological outcomes of patients treated in Denmark between 2010 and 2022. Method. We present a retrospective review of a national cohort comprising 27 patients. The indications were 13 cases of Ewing sarcoma, 12 cases of osteosarcoma, and 2 cases of giant cell tumor. The median age at surgery was 16 years (range: 2-39), and the median follow-up was 82 months (range: 12-138). Patients were analyzed overall and stratified into upper and lower extremity groups based on tumor location. Result. The primary rate of graft union was 63%, and after secondary procedures, the overall rate of graft union was 67%, with a median time to union of 13 months (range: 7-29). The reoperation rate was 74%, while the rate of limb salvage was 93%, with two patients undergoing amputation during follow-up. The 5-year overall survival rate was 81% (95% CI: 61-92). Patients with upper extremity tumors were more likely to attain graft union (92% vs. 47%, p=0.02) and less likely to undergo multiple reoperations (17% vs 60%, p=0.047) compared to patients with lower extremity tumors. Conclusion. Vascularized fibula grafting remains a valuable option in limb salvage surgery with acceptable long-term outcomes. However, especially in lower extremity cases, a low rate of graft union and multiple reoperations are to be expected

Previous scientific studies have highlighted how coupling is an important element affecting total hip arthroplasty's survival. This study aims to evaluate whether metal-on-metal (MOM) coupling could be a statistically significant risk factor. The data from the regional joint registry (Registro dell'Impiantologia Protesica Ortopedica, RIPO) was used for analysis. The data collection accuracy of this registry was 97.2% in 2017. We retrospective evaluate all MOM total hip arthroplasties (THAs) implanted in our department between January 01st 2000 and December 31st 2011. We used a control group composed by all other prosthesis implanted in our Department in the same time lapse. We registered 660 MOM THAs. Mean age of patients was 66.9 years. 603 patients have a >36mm head, while 78 a <36 mm one. Neck modularity was present in half of patients. 676 implants were cementless. We registered 69 revisions, especially due to aseptic mobilization (16 THAs), implant breakage (9 THAs) and periprosthetic fracture (6 THAs). The MOM THAs overall Kaplan-Meier survival rate was 87.2 at 15 years, and the difference between MOM THAs and other implants two curves is statistically significant (p<0.05). Male sex is a significant risk factors. Further evaluations are in progress to establish the presence of any additional risk factors. We think weight and/or BMI may be included in this category. Our study confirms the data currently present in the literature regarding a lower survival of metal-on-metal hip prostheses. The male sex is a statistically significant risk factor (p<0.05), while age, head size and modularity of the prosthetic neck are not statistically significant (p>0.05). Any new finds will be presented at the congress venue

Results in patients undergoing total hip arthroplasty (THA) for femoral head osteonecrosis (ON) when compared with primary osteoarthritis (OA) are controversial. Different factors like age, THA type or surgical technique may affect outcome. We hypothesized that patients with ON had an increased revision rate compared with OA. We analysed clinical outcome, estimated the survival rate for revision surgery, and their possible risk factors, in two groups of patients. In this retrospective cohort analysis of our prospective database, we assessed 2464 primary THAs implanted between 1989 and 2017. Patients with OA were included in group 1, 2090 hips; and patients with ON in group 2, 374 hips. In group 2 there were more men (p<0.001), patients younger than 60 years old (p<0.001) and with greater physical activity (p<0.001). Patients with lumbar OA (p<0.001) and a radiological acetabular shape type B according to Dorr (p<0.001) were more frequent in group 1. Clinical outcome was assessed according to the Harris Hip Score and radiological analysis included postoperative acetabular and femoral component position and hip reconstruction. Kaplan-Meier survivorship analysis was used to estimate the cumulative probability of not having revision surgery for different reasons. Univariate and multivariate Cox regression models were used to assess risk factors for revision surgery. Clinical improvement was better in the ON at all intervals. There were 90 hips revised, 68 due to loosening or wear, 52 (2.5%) in group 1, and 16 (4.3%) in group 2. Overall, the survival rate for revision surgery for any cause at 22 years was 88.0 % (95% CI, 82-94) in group 1 and 84.1% (95% CI, 69 – 99) in group 2 (p=0.019). Multivariate regression analysis showed that hips with conventional polyethylene (PE), compared with highly-cross linked PEs or ceramic-on-ceramic bearings, (p=0.01, Hazard Ratio (HR): 2.12, 95% CI 1.15-3.92), and cups outside the Lewinnek´s safe zone had a higher risk for revision surgery (p<0.001, HR: 2.57, 95% CI 1.69-3.91). Modern highly-cross linked PEs and ceramic-on-ceramic bearings use, and a proper surgical technique improved revision rate in patients undergoing THA due to ON compared with OA

Although cemented fixation provides excellent results in primary total hip replacement (THR), particularly in patients older than 75 years, uncemented implants are most commonly used nowadays. We compare the rate of complications, clinical and radiological results of three different designs over 75-years-old patients. 433 hips implanted in patients over 75 years old were identified from our Local Joint Registry. Group A consisted of 139 tapered cemented hips, group B of 140 tapered grit-blasted uncemented hips and group C of 154 tapered porous-coated uncemented hips. A 28 mm femoral head size on polyethylene was used in all cases. The mean age was greater in group A and the physical activity level according to Devane was lower in this group (p<0.001 for both variables). Primary osteoarthritis was the most frequent diagnoses in all groups. The radiological acetabular shape was similar according to Dorr, however, an osteopenic-cylindrical femur was most frequently observed in group A (p<0.001). The pre- and post-operative clinical results were evaluated according to the Merle-D'Aubigne and Postel scale. Radiological cup position was assessed, including hip rotation centre distance according to Ranawat and cup anteversion according to Widmer. We also evaluated the lever arm and height of the greater trochanter distances and the stem position. Kaplan-Meier analysis was done for revision for any cause and loosening. The hip rotation centre distance was greater and the height of the greater trochanter was lower in group B (p=0.003, p<0.001, respectively). The lever arm distance was lower in group C (p<0.001). A varus stem position was more frequently observed in group B (p<0.001). There were no intra- or post-operative fractures in group A, although there were five intra-operative fractures in the other groups plus two post-operative fractures in group B and four in group C. The rate of dislocation was similar among groups and was the most frequent cause for revision surgery (8 hips for the whole series). The mean post-operative clinical score improved in all groups. The overall survival rate for revision for any cause at 120 months was 88.4% (95% CI 78.8–98), being 97.8% (95% CI 95.2–100) for group A, 81.8% (95% CI 64.8–98.8) for group B and 95.3% (95% CI 91.1–99.6) for group C (log Rank: 0.416). Five hips were revised for loosening. The overall survival rate for loosening at 120 months was 91.9% (95% CI 81.7–100), being 99.2%(95% CI 97.6–100) for group A, 85.5 (95% CI 69.9 −100) for group B and 100% for group C (Log Rank 0.093). Despite a more osteopenic bone in the cemented group, the rate of peri-prosthetic fractures was higher after uncemented THR in patients older than 75 years. Although the overall outcome is good with both types of fixation, the post-operative reconstruction of the hip, which might be more reliable after cemented fixation, may affect the rate of complications in this population

Summary Statement. RANK is expressed in 18% of human osteosarcomas and is likely to provide additional prognostic information for clinical purposes in osteosarcoma patients at the time of diagnosis. Introduction. The receptor activator of nuclear factor kappa (RANK), a member of the tumor necrosis factor family, is activated by its ligand and regulates the differentiation of osteoclasts and dendritic cells. Local growth of osteosarcoma involves destruction of the host bone by osteoclasts and proteolytic mechanisms. Although prognosis of osteosarcoma has been improved by chemotherapy during the last decades, the problem of non responders and the lack of prognostic markers remains. It is the aim of this study to investigate the prognostic and predictive value of RANK expression in human osteosarcoma. Patients & Methods. The expression of RANK was examined immunohistochemically in biopsies of 43 patients (mean age 25.7 years) with high grade osteosarcoma and the results were correlated with histologic response to chemotherapy, disease free and overall survival. Tumors with more than 40% positive osteosarcoma cells were scored positive. Results. In 8 of 43 (18%) osteosarcoma specimens RANK expression could be dedected, the rest were negative. RANK expression showed a statistically significant correlation with overall survival of patients. 7/8 patients with RANK expressing tumours died, whereas only one in the negative group (88% in RANK positive tumours versus 37%; p<0.05). No significant difference was found when comparing RANK expression status with response to chemotherapy; 50% had a poor and 50% had a good response in RANK positive and 30,3% had a good and 69,7% had a bad response in RANK negative osteosarcomas. The appearance of metastases did not correlate with RANK expression status (37,5% metastases in RANK positive tumours versus 28,6% in negative). Discussion/Conclusion. In conclusion our findings suggest that RANK is likely to provide additional prognostic information for clinical purposes in osteosarcoma patients at the time of diagnosis

Neck modularity has been proposed to improve THA accuracy, thanks to the close restoration of anatomy, however it has been associated with issues like early breakages or corrosion. Our Hospital has been using neck modularity since the 90s, so we analyzed retrospectively implants performed between January 2000 and December 2014. The minimum follow up was 1Y. The cohort was composed of 1,033 THAs or 951 patients (82 bilateral), of which 643 females and 390 males. Average patient age was 67.7Y. THA indications were primary Osteoarthritis (80.9%), Fracture (9.0%), Congenital Dysplasia or Congenital Luxation (4.2%), Osteonecrosis (3.2%), other causes (2,7%). The stems used were all cementless, 381 anatomically shaped (36.9%), 635 straight (61.5%), 17 short MIS (1.6%). All necks used were made of Titanium alloy. 419 implants (40.5%) were manufactured by Wright Medical, while 614 (59.5%) were produced by Adler Ortho. A total of 37 revisions has been reported, mainly due to periprosthetic fractures (32.4%), luxation (24.3%), implant mobilization (18.9%) and implant breakage (16.2%). We have recorded 3 modular neck breakages. 4 patients required re-revisions, because of luxations (3) and neck breakage (1). The overall survival rate was 96.4%. We did not observe any component corrosion, probably thanks to the exclusive use of Titanium necks. We had a neck breakages rate of 0.29% and a luxation rate of 0.87%, lower than normally reported in the literature. In conclusion, our experience suggests as neck modularity could be a safe and effective way to reconstruct the proximal femur in THA patients

Osteosarcoma (OS) is a highly malignant primary tumor frequently occurring in children and adolescents. The mainstay of therapy is neoadjuvant chemotherapy and surgical removal of the lesion yielding a 50–70% of 5-year survival rate. Unfortunately, chemotherapy is currently unable to induce complete tumor necrosis leaving residual tumor cells free to metastasize or recidivate, thus resulting in a 30% mortality. The major limitation in those patients is the development of multidrug resistance (MDR) and the low water solubility of drugs such as Paclitaxel (PTX) that is in fact not included in the majority of chemotherapy protocols for OS treatment. We thus hypothesized to prevent the emergence of MDR and obtain significant tumor reduction, by engineering innovative nanoparticles (NPs) able to vehiculate the PTX and induce a dual synergic action: the cytostatic effect of PTX and the cytotoxicity generated by reactive oxygen species produced from light triggered photoactivation (PDT) of Chlorin e6 photosensitizer. To further improve the efficacy and reduce the side effects of NPs systemic administration, Mesenchymal Stromal Cells (MSC) are used as a “Trojan horse” to deliver the NPs directly to tumor cells, taking advantage of MSC ability to selectively recognize and efficiently engraft in OS tumor stroma. HSA were conjugated with photosensitizer Ce6 and the functionalized protein was used to produce PTX loaded nanoparticles through desolvation technique and drug-induced protein self-assembly (PTX-Ce6@HSA NP). Human MSC lines, isolated from the Bone marrow (BM) of different donors, were then loaded with different dosages of nanoparticles and their ability to internalize and transport the NPs, migrate and induce cytotoxic ROS upon light treatment were tested in in vitro cultures. Preliminary results showed that MSC efficiently internalize PTX-Ce6@HSA NPs and the photosensitizer Ce6 remains active inside the cells for at least 3 days after loading. Electron microscopy performed onto loaded MSC showed that NPs internalization take places via clathrin mediated transport, whereas HPLC analysis demonstrated a release kinetics of PTX mediated by exocytosis. Finally, PTX-Ce6@HSA NPs loaded MSC co-cultured with the OS tumor cell line SaOS-2 showed a significant tumor cell growth reduction. So fare, advances in drug delivery have failed to produce specific tool to improve the overall survival of OS patients. However, given our preliminary in vitro data we believe that the proposed multimodal therapy will minimize the side effects of the systemic chemotherapy and enhance the efficacy through the synergic effect of PTX and PDT. In the future, our strategy could be intended as an innovative co-adjuvant approach for OS treatment to be performed right before surgery to eliminate residual tumors cells after tumor mass removal

Summary Statement. Survivin is a member of the inhibitor of apoptosis family, which may contribute to the progression of human MFH via inhibiting the mitochondrial apoptosis, and may be considered as a potent therapeutic target for the treatment of human MFH. Introduction. Survivin is a member of the inhibitor of apoptosis (IAP) family, which usually expresses in the embryonic lung and fetal organs in the developmental stages, but is undetectable in normal adult tissues other than thymus, placenta, CD34. +. stem cells, and basal colonic epitherial cells. However, several studies reported that survivin is highly expressed in various human malignancies, including sarcomas, and increased expression of survivin is an unfavorable prognostic marker correlating with decreased overall survival in cancer patients. We have previously reported that survivin was strongly expressed in human malignant fibrous histiocyoma (MFH), however, the roles of survivin in human MFH have not been studied. The aim of this study was to evaluate the effect of survivin inhibition on apoptotic activity in human MFH cells. Methods. Nara-H, a human MFH cell line which expresses the high levels of survivin, was used in this study. Cells were cultured in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin at 37°C in a humidified atmosphere containing 5% CO. 2. To evaluate the effect of survivin inhibition on MFH cell apoptosis, cells were transfected with either a survivin specific siRNA (survivin-siRNA) or a non-specific control siRNA (control-siRNA) by lipofection method. After siRNA transfection, the efficiency of siRNA knockdown of survivin was assessed by quantitative real time PCR. Expressions of apoptosis-related proteins, such as caspase-3, caspase-9 and PARP, were assessed by immunoblot analysis, and the apoptotic activity was evaluated by flow cytometric analysis. Results. Transfection of survivin-siRNA strongly suppressed the expression of survivin compared with control-siRNA. Immunoblot analyses revealed that expressions of cleaved forms of caspase-3, caspase-9 and PARP were increased in survivin-siRNA transfected cells, while the expressions were barely detected in control cells. In flow cytometric analysis, the number of apoptotic cells was significantly increased in survivin-siRNA transfected cells compared with that in control cells. Discussion/Conclusion. Previous studies revealed that survivin regulates the mitochondrial apoptotic pathway, and that overexpression of survivin is associated with tumor growth, progression, and resistance to conventional targeted anticancer agents in various human malignancies. In the current study, we demonstrated that siRNA knockdown of survivin induced the cleavage of caspase-3, caspase-9 and PARP, and increased the apoptotic activity in human MFH cells. The findings in this study strongly suggest that survivin may contribute to the progression of human MFH via inhibiting the mitochondrial apoptosis in human MFH, and may be considered as a potent therapeutic target for the treatment of human MFH

We investigated the rates of expression of bone morphogenetic protein-2 (BMP-2) in 29 adult patients with high-grade malignant fibrous histiocytoma of soft tissue, using the BMP-2-specific monoclonal antibody, AbH3b2/17, and found that they ranged from 1.9% to 78.9%. The survival at five years of the groups expressing high (≥30%) and low (< 30%) levels of BMP-2 was 85.7% and 36.3%, respectively. Multivariable analysis showed that only BMP-2 had prognostic significance for continuous disease-free survival and for overall survival (p < 0.05). Our findings indicate that over-expression of BMP-2 in malignant fibrous histiocytoma of soft tissue is the most reliable prognostic indicator of the parameters assessed

In vivo animal experimentation has been one of the cornerstones of biological and biomedical research, particularly in the field of clinical medicine and pharmaceuticals. The conventional in vivo model system is invariably associated with high production costs and strict ethical considerations. These limitations led to the evolution of an ex vivo model system which partially or completely surmounted some of the constraints faced in an in vivo model system. The ex vivo rodent bone culture system has been used to elucidate the understanding of skeletal physiology and pathophysiology for more than 90 years. This review attempts to provide a brief summary of the historical evolution of the rodent bone culture system with emphasis on the strengths and limitations of the model. It encompasses the frequency of use of rats and mice for ex vivo bone studies, nutritional requirements in ex vivo bone growth and emerging developments and technologies. This compilation of information could assist researchers in the field of regenerative medicine and bone tissue engineering towards a better understanding of skeletal growth and development for application in general clinical medicine.

Cite this article: A. A. Abubakar, M. M. Noordin, T. I. Azmi, U. Kaka, M. Y. Loqman. The use of rats and mice as animal models in ex vivo bone growth and development studies. Bone Joint Res 2016;5:610–618. DOI: 10.1302/2046-3758.512.BJR-2016-0102.R2.

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.