Undiagnosed synovial hemangioma of the knee: a case report.

BACKGROUND: Synovial hemangioma of the knee is a rare benign tumor. Very rarely, the growth of bone is affected by long-term neglect of an intra-articular tumor. Our patient had not only various clinical symptoms but also dysplasia of the femoral bone. In this report, we aimed to raise awareness to prevent various disorders arising from an unnoticed or untreated hemangioma occurring within the knee joint. CASE PRESENTATION: Our patient was a 41-year-old Japanese man who had had occasional discomfort in the right knee since elementary school. Although he had undergone radiography at several hospitals since childhood, no issues were reported; subsequently, he consulted our hospital. We performed magnetic resonance imaging and discovered a mass. The mass was homogeneous with low intensity on T1-weighted sequences and high intensity on T2-weighted sequences adjacent to the medial femoral condyle. The shape of the medial femoral condyle presented with a concavity in axial images, with irregular margins from the patellofemoral joint to the medial femoral condyle. Moreover, by using magnetic resonance angiography, we discovered a second mass. We decided to perform open surgery to achieve complete excision. Histological examination indicated a synovial hemangioma involving a cavernous hemangioma and irregular arteriovenous connections originating from the subsynovial tissue. The patient became asymptomatic after surgery, with no recurrence for more than 4 years. CONCLUSIONS: Synovial hemangioma is rare and difficult to diagnose in outpatient examinations because radiography has a limited diagnostic capacity. Magnetic resonance imaging and angiography are very useful. Nontreatment of intra-articular hemangiomas may lead to dysplasia of the bone and various clinical symptoms. Early complete excision may be instituted to reduce these risks of hemarthrosis.

Advantages of creation of holes and removal of air in artificial bone for early bone formation when used artificial bone as a gap filler in open wedge high tibial osteotomy.

Recently, many facilities perform open wedge high tibial osteotomy (OWHTO) using artificial bone as a gap filler. However, there are many cases in which artificial bone is used without a clear purpose. We recommend a surgical technique to promote early synostosis between artificial bone and recipient bone due to mechanical support especially in the early stage after OWHTO. At our hospital, beta-tricalcium phosphate (ß-TCP) with 60% porosity is used in OWHTO. Initially, a wedge-shaped block-type ß-TCP, as large as possible, was inserted into the gap. However, from the standpoint of initial mechanical support, we changed the artificial bone size and created intentional holes. Furthermore, we removed air bubbles from ß-TCP. We evaluated the synostosis on the basis of clinical results and diagnostic imaging. As a result of creating holes and removing air from the artificial bone, a trend toward faster synostosis was noted, especially at the early stage. No adverse events such as tibial plateau fracture, lateral cortical fracture, plate and screw failure and correction loss due to reducing the size of the artificial bone occurred, but placement of the artificial bone in contact with cortical bone and surface contact installation with the recipient bone tissue was important. When using artificial bone in OWHTO, holes formation and removal of air from the artificial bone are recommended for faster synostosis between artificial bone and recipient bone in the early stage after surgery. Artificial bone should be used, with attention to its positioning and shape, for efficient mechanical support.

Reg Gene Expression in Periosteum after Fracture and Its In Vitro Induction Triggered by IL-6.

The periosteum is a thin membrane that surrounds the outer surface of bones and participates in fracture healing. However, the molecular signals that trigger/initiate the periosteal reaction are not well established. We fractured the rat femoral bone at the diaphysis and fixed it with an intramedullary inserted wire, and the expression of regenerating gene (Reg) I, which encodes a tissue regeneration/growth factor, was analyzed. Neither bone/marrow nor muscle showed RegI gene expression before or after the fracture. By contrast, the periosteum showed an elevated expression after the fracture, thereby confirming the localization of Reg I expression exclusively in the periosteum around the fractured areas. Expression of the Reg family increased after the fracture, followed by a decrease to basal levels by six weeks, when the fracture had almost healed. In vitro cultures of periosteal cells showed no Reg I expression, but the addition of IL-6 significantly induced Reg I gene expression. The addition of IL-6 also increased the cell number and reduced pro-apoptotic gene expression of Bim. The increased cell proliferation and reduction in Bim gene expression were abolished by transfection with Reg I siRNA, indicating that these IL-6-dependent effects require the Reg I gene expression. These results indicate the involvement of the IL-6/Reg pathway in the osteogenic response of the periosteum, which leads to fracture repair.

Bizarre soft tissue tumour on the gluteus muscle: a case report of spontaneous chronic expanding haematoma.

Chronic expanding haematoma (CEH) is rare and refers to a gradually increasing haematoma that is not absorbed after surgery and trauma. This report highlights unusual mass occurring on the gluteus muscle, and the aim is to indicate the diagnostic method. It is necessary to consider the mechanism of the occurrence and to know the characters of CEH. The patient was a 51-year-old man who had noticed a soft mass on his right hip. The mass had gradually increased to 10 cm in size over the year. CT images revealed a haematoma. However, MRI showed a rare biphasic fluid-fluid layer inside the mass and indicated a different pattern compared with that of a normal haematoma. Because the mass was affecting the patient's social life, and the diagnosis was difficult to confirm, surgical treatment was elected. Intraoperatively, the mass contained a large amount of a brown mud-like substance and showed the bizarre appearance inside. The mass was diagnosed as CEH based on both the clinical findings and the histopathological diagnosis. The patient had no traumatic event and no previous surgery. In the absence of the clinical history and the unique imaging findings, it was difficult to diagnose the mass as CEH. It is important to clarify a patient's underlying disease, history, and lifestyle and to consider any correlation between the mass location and the patient's condition carefully. Considering the character of the mass and the lack of a preoperative definitive diagnosis, we recommend performing complete surgical resection.

Calcium Concentration in Culture Medium as a Nondestructive and Rapid Marker of Osteogenesis.

Artificial bones made of ß-tricalcium phosphate (ß-TCP) combined with bone marrow-derived mesenchymal stromal cells (BM-MSCs) are used for effective reconstruction of bone defects caused by genetic defects, traumatic injury, or surgical resection of bone tumors. However, the selection of constructs with high osteogenic potential before implantation is challenging. The purpose of this study was to determine whether the calcium concentration in BM-MSC culture medium can be used as a nondestructive and simple osteogenic marker for selecting tissue-engineered grafts constructed using ß-TCP and BM-MSCs. We prepared three cell passages of BM-MSCs derived from three 7-week-old, male Fischer 344 rats; the cells were cultured in osteoinductive medium in the presence of ß-TCP for 15 days. The medium was replaced with fresh medium on day 1 in culture and subsequently changed every 48 h; it was collected for measurement of osteocalcin secretion and calcium concentration by enzyme-linked immunosorbent assay and X-ray fluorescence spectrometry, respectively. After cultivation, the constructs were implanted subcutaneously into the backs of recipient rats. Four weeks after implantation, the alkaline phosphatase (ALP) activity and osteocalcin content of the constructs were measured. A strong inverse correlation was observed between the calcium concentration in the medium and the ALP activity and osteocalcin content of the constructs, with Pearson's correlation coefficients of 0.92 and 0.90, respectively. These results indicate that tissue-engineered bone with high osteogenic ability can be selected before implantation based on low calcium content of the culture medium, resulting in successful bone formation after implantation. This nondestructive, simple method shows great promise for assessing the osteogenic ability of tissue-engineered bone.

Osteogenic Matrix Cell Sheets Facilitate Osteogenesis in Irradiated Rat Bone.

Reconstruction of large bone defects after resection of malignant musculoskeletal tumors is a significant challenge in orthopedic surgery. Extracorporeal autogenous irradiated bone grafting is a treatment option for bone reconstruction. However, nonunion often occurs because the osteogenic capacity is lost by irradiation. In the present study, we established an autogenous irradiated bone graft model in the rat femur to assess whether osteogenic matrix cell sheets improve osteogenesis of the irradiated bone. Osteogenic matrix cell sheets were prepared from bone marrow-derived stromal cells and co-transplanted with irradiated bone. X-ray images at 4 weeks after transplantation showed bridging callus formation around the irradiated bone. Micro-computed tomography images at 12 weeks postoperatively showed abundant callus formation in the whole circumference of the irradiated bone. Histology showed bone union between the irradiated bone and host femur. Mechanical testing showed that the failure force at the irradiated bone site was significantly higher than in the control group. Our study indicates that osteogenic matrix cell sheet transplantation might be a powerful method to facilitate osteogenesis in irradiated bones, which may become a treatment option for reconstruction of bone defects after resection of malignant musculoskeletal tumors.

Comparison of gene expression profiling in sarcomas and mesenchymal stem cells identifies tumorigenic pathways in chemically induced rat sarcoma model.

Mesenchymal stem cells (MSCs) are believed to be the cell of origin for most sarcomas including osteosarcoma and malignant fibrous histiocytoma (MFH/UPS). To identify the signaling pathways involved in sarcoma pathogenesis, we compared gene expression profiles in rat osteosarcoma and MFH cells with those in syngeneic rat MSCs. Analysis of genes that characterize MSCs such as CD44, CD105, CD73, and CD90 showed higher expression in MSCs compared to sarcomas. Pathways involved in focal and cell adhesion, cytokine-cytokine receptors, extracellular matrix receptors, chemokines, and Wnt signaling were down-regulated in both sarcomas. Meanwhile, DNA replication, cell cycle, mismatch repair, Hedgehog signaling, and metabolic pathways were upregulated in both sarcomas. Downregulation of p21(Cip1) and higher expression of CDK4-cyclinD1 and CDK2-cyclinE could accelerate cell cycle in sarcomas. The current study indicated that these rat sarcomas could be a good model for their human counterparts and will provide the further insights into the molecular pathways and mechanisms involved in sarcoma pathogenesis.

Early fixation of cobalt-chromium based alloy surgical implants to bone using a tissue-engineering approach.

To establish the methods of demonstrating early fixation of metal implants to bone, one side of a Cobalt-Chromium (CoCr) based alloy implant surface was seeded with rabbit marrow mesenchymal cells and the other side was left unseeded. The mesenchymal cells were further cultured in the presence of ascorbic acid, ß-glycerophosphate and dexamethasone, resulting in the appearance of osteoblasts and bone matrix on the implant surface. Thus, we succeeded in generating tissue-engineered bone on one side of the CoCr implant. The CoCr implants were then implanted in rabbit bone defects. Three weeks after the implantation, evaluations of mechanical test, undecalcified histological section and electron microscope analysis were performed. Histological and electron microscope images of the tissue engineered surface exhibited abundant new bone formation. However, newly formed bone tissue was difficult to detect on the side without cell seeding. In the mechanical test, the mean values of pull-out forces were 77.15 N and 44.94 N for the tissue-engineered and non-cell-seeded surfaces, respectively. These findings indicate early bone fixation of the tissue-engineered CoCr surface just three weeks after implantation.

Mesenchymal stem cells promote tumor engraftment and metastatic colonization in rat osteosarcoma model.

Although mesenchymal stem cells (MSCs) are considered to be the cells of origin for most sarcomas, the role of MSCs as a source of tumor stroma is not fully understood in this tumor type. The current study investigated whether MSCs affect the tumor growth and metastatic ability in rat osteosarcoma model. Results from subcutaneous co-implantation of rat osteosarcoma COS1NR cells, established in our laboratory, with rat MSCs isolated from femur bone marrow showed that the incidence of tumor formation and tumor growth rate was higher until 5 weeks compared to COS1NR cell inoculation alone. However, no difference was observed in tumor growth afterwards and in the number of metastatic nodules at 9 weeks (0.75 vs. 1.2). Intravenous MSC injection at weeks 3 and 5 after subcutaneous inoculation of COS1NR cells significantly increased the number of lung nodules in the group with MSC injection compared to the group without MSC injection (17.33 vs. 2.0), while no difference was observed in subcutaneous tumor growth between those groups. Pathway analysis from gene expression profile identified that genes involved in focal adhesion, cytokine-cytokine receptor and extracellular matrix-receptor pathways such as CAMs (ICAM and VCAM)-integrins were highly expressed in MSCs, possibly participating in the tumor progression of osteosarcoma. These results suggest that MSCs could provide a source of microenvironments for osteosarcoma cells, and might enhance the ability of settlement and colonization which lead to early onset of growth and metastasis, possibly through their activated pathways interaction.

Osteogenic activity of bone marrow-derived mesenchymal stem cells (BMSCs) seeded on irradiated allogenic bone.

Allogenic bone grafting, a technique used in orthopaedic surgery, has several problems, including low osteogenic activity. To overcome the problem, this study aimed to determine whether in vivo osteogenesis could be enhanced using allogenic irradiated bone grafts after seeding with autologous bone marrow-derived mesenchymal stem cells (BMSCs). The allogenic bone cylinders were extracted from ACI rats and sterilized by irradiation. Donor BMSCs were obtained from fresh Fischer 344 (F344) rat bone marrow by cell culture. The allogenic bone with or without BMSCs were transplanted subcutaneously into syngeneic F344 rats. At 4 weeks after transplantation, high alkaline phosphatase (ALP) activity, bone-specific osteocalcin mRNA expression and newly formed bone were detected in the allogenic bone with BMSCs. The origin of the newly formed bone was derived from cultured donor BMSCs. However, none of these identifiers of osteogenesis were detected in either the fresh or the irradiated allogenic bone without BMSCs. These results indicate the availability of autologous BMSCs to heighten the osteogenic response of allogenic bone. Our present tissue-engineering method might contribute to a wide variety of allogenic bone grafting techniques in clinical settings.

Secretory osteocalcin as a nondestructive osteogenic marker of tissue-engineered bone.

BACKGROUND AND PURPOSE: The constructs of mesenchymal stem cells and ceramics form bone tissue after implantation. Therefore, the constructs can include cultured bone (tissue-engineered bone) as bone grafts. However, the selection of constructs, prior to implantation, with high osteogenic potential is still difficult. We used a rat model to measure the secretory osteocalcin level in culture medium to verify that monitoring osteocalcin levels enables the selection of constructs with high osteogenic potential. METHODS: We prepared constructs of rat hydroxyapatite/cells and used different cell passages of P-1 and P-3 as well as different cell numbers: 1 × 10(5) and 1 × 10(6) cells/ml suspension. These constructs were cultured for 14 days under osteoinductive or nonosteoinductive conditions and implanted subcutaneously in the recipient rat. Secretory osteocalcin in the culture medium was measured using an enzyme-linked immunosorbent assay system during the culture period until day 14, and the osteocalcin content of the harvested construct at 4 weeks was also measured. RESULTS AND CONCLUSION: All constructs except the hydroxyapatite/P-3 construct showed abundant bone formation by histology and both high secretory osteocalcin level in the medium and high osteocalcin content after implantation. Our study revealed that secretory osteocalcin level in vitro was related to osteocalcin content in vivo. The study clearly showed that measuring secretory osteocalcin is a nondestructive method of assessing the osteogenic potential of tissue-engineered bone. One can choose tissue-engineered bone with high osteogenic potential by integrating secretory osteocalcin measurement into the process of bone-tissue regeneration.

Popliteal soft tissue tumor associated with hamstring injury.

We report herein a very rare case of semitendinosus tear that formed a tumor in the popliteal region after not having recovered as a result of only being instructed to rest. The soft tissue tumor was discovered on ultrasonography 4 months after pain and sensation of discomfort appeared in the popliteal region. We considered this symptom as dependent on the presence of the tumor and selected surgical treatment. Intraoperatively, this soft tissue tumor was connected with the pes anserinus by tendinous tissue. On pathological examination, the soft tissue tumor was diagnosed as skeletal muscle showing necrosis. From imaging, operative findings, and pathological diagnosis, this was considered to represent a rare case in which myorrhexis developed into a soft tissue tumor in the popliteal region after a semitendinosus tear remained unhealed and was neglected over the long term. Although we had trouble confirming a diagnosis and treatment procedure, we were able to acquire good results with surgical treatment. This is, to the best of our knowledge, the first report of damaged semitendinosus becoming a soft tissue tumor in the popliteal region after long-term neglect without healing.

Scaffold-free cell sheet injection results in bone formation.

We previously reported a new cell transplantation method in which mesenchymal stem cells (MSCs) were cultured as cell sheets. The cultured MSC sheets showed high alkaline phosphatase (ALP) activities and osteocalcin (OC) contents. In the present study, we transplanted such sheets by injection to assess whether the injectable MSC sheets could form bone tissue at subcutaneous sites. At 4 weeks after the subcutaneous injection, the injected areas showed hard mass formation. Each mass consisted of newly formed bone, as evaluated by radiographic, histological and gene expression analyses as well as three-dimensional computed tomography (3D-CT). Histological analyses revealed extracellular bone matrix together with osteocytes and active osteoblasts. Real-time PCR analyses showed high ALP and OC mRNA expressions. We also injected the cell sheets into dead bone to determine whether the lost osteogenic potential could be rescued, and histological analyses revealed that the injected cell sheets supplied osteogenic potential to the dead bone. The present study clearly indicates that osteogenic MSC sheets can be transplanted via injection through a needle and that bone formation results in the injected areas. Owing to its usage of a needle for fabrication of in vivo bone tissue, this injection method can be applied as a minimally invasive approach for hard tissue reconstruction.

Bone marrow-derived mesenchymal cells can rescue osteogenic capacity of devitalized autologous bone.

In clinical cases, many orthopaedists have been troubled with bone fragility, such as fractures after devitalization therapy for bone tumour, pathological fractures and metastatic tumours. The aim of this study was to determine whether loss of osteogenic capacity of devitalized autologous bones can be rescued using cultured bone marrow-derived mesenchymal cells. A devitalized bone model was produced from rat femur by irradiation and three groups were prepared: intact bone, irradiated bone and irradiated bone combined with cultured mesenchymal cells. Each bone was transplanted subcutaneously into a syngeneic rat. At 2 or 4 weeks after transplantation, biochemical analyses [alkaline phosphatase (ALP) activity and osteocalcin mRNA expression] and histological measurement were performed. Moreover, we verified the origin of newly formed bone, using the sex-determining region Y (sry) gene as a marker to distinguish between donor and recipient. In both intact bone and irradiated bone with mesenchymal cells, ALP activity and osteocalcin mRNA expression were detected and living osteoblasts together with newly formed bone were clearly seen histologically. Furthermore, analysis of the origin of de novo formed bone indicated that newly formed bone in irradiated bone with mesenchymal cells was derived from cultured bone marrow-derived mesenchymal cells. These results proved that the osteogenic capacity of devitalized autologous bone can be rescued using tissue-engineering techniques. This procedure should contribute to various clinical treatments, such as local metastatic tumours, pathological fracture after devitalization therapy and reconstruction after wide-margin tumour resection. The benefits would be applicable to all types of devitalized bone.

Long-term clinical and radiographic follow-up of total resection for discoid lateral meniscus.

PURPOSE: The purpose of this retrospective study was to analyze the effects of age at surgery on the clinical results, arthritic progression, and changes in lower limb alignment during long-term follow-up of total resection for discoid lateral meniscus. METHODS: A total of 37 knees in 32 patients were reviewed as a whole group and also after division into the following 3 groups according to the age at surgery: group A, 11 knees in patients aged less than 20 years; group B, 13 knees in patients aged 20 to 39 years; and group C, 13 knees in patients aged greater than 39 years. The mean age at surgery was 31.2 years (range, 9 to 52 years), and the mean length of follow-up was 14.5 years (range, 10 to 22.4 years). RESULTS: According to the scale of Ikeuchi, clinically excellent or good results at the final follow-up were found in 84% of the knees in the whole group, 91% of those in group A, 85% of those in group B, and 77% of those in group C. Moderate or severe narrowing of the lateral joint space was seen in 11% of the knees in the whole group, 0% of those in group A, 8% of those in group B, and 23% of those in group C. In an analysis of lower limb alignment, statistically significant postoperative lateral shifts of Mikulicz's mechanical axis were observed in groups B and C. CONCLUSIONS: The clinical results were satisfactory and the postoperative arthritic changes were mild in patients aged under 40 years. However, it is necessary to carefully monitor arthritic progression in patients who are aged over 19 years and have valgus deformities, because most of the knees in patients above this age became more valgus postoperatively. LEVEL OF EVIDENCE: Level IV, prognostic case series.

Early bone in-growth ability of alumina ceramic implants loaded with tissue-engineered bone.

To enhance early bonding of an alumina ceramic implant to bone, we evaluated a method of seeding the implant surface with bone marrow mesenchymal cells that differentiated to osteoblasts and bone matrix prior to implantation. The usefulness of the method was evaluated in Japanese white rabbits. In our study, an alumina ceramic test piece loaded with differentiated osteoblasts and bone matrix by a tissue engineering technique was implanted into rabbit bones. Three weeks after the procedure, evaluation of mechanical bonding and histological examination were performed. Histological examination of the noncell-loaded implant surfaces showed no bone infiltration into the implant gap. However, the cell-loaded implant surfaces exhibited new bone infiltration into the implant gap with mechanical bonding. In the mechanical test, the average failure load was 0.60 kgf for the noncell-loaded side and 1.49 kgf for the cell-loaded side. Preculturing mesenchymal cells on the surface of the alumina ceramic prior to implantation increased the debonding strength by two and half times. The present findings indicate early bonding between the implant and bone three weeks after the procedure.