Kumi Orita

Background/Objective: Anti-tumor effects of plasma-activated medium (PAM) were demonstrated using various malignant tumors. However, the anti-tumor effect of PAM on synovial sarcoma remains unclear. Therefore, we aimed to investigate the anti-tumor effects of PAM on synovial sarcoma and its underlying mechanisms, focusing on the quantitative analyses of both intracellular reactive oxygen species (ROS) and cell apoptosis. Methods: The human synovial sarcoma cell line HS-SY-II was used to investigate the cell viability after PAM treatment. We investigated the anti-tumor effects and side effects of local PAM injection in a synovial sarcoma xenograft murine model. Moreover, we observed PAM-induced intracellular ROS accumulation and cell apoptosis and assessed the involvement of intracellular ROS in the anti-tumor effects of PAM using an intracellular ROS scavenger. Results: PAM significantly decreased the viability of synovial sarcoma cells compared with untreated Dulbecco’s modified Eagle medium. Local PAM injection into a synovial sarcoma xenograft murine model significantly suppressed tumor growth, including tumor volume (p < 0.001) and weight (p = 0.031), without side effects. Regarding anti-tumor mechanisms, PAM induced significant cell apoptosis and intracellular ROS accumulation (p < 0.001). The intracellular ROS scavenger significantly inhibited the anti-tumor effect of PAM (p < 0.001). Conclusions: We confirmed the anti-tumor effects of PAM on synovial sarcoma in vitro and in vivo, as well as the absence of side effects. The underlying mechanism was suggested to involve cell apoptosis induced by intracellular ROS accumulation. Considering the various clinical issues associated with the existing treatments of synovial sarcoma, PAM is a promising new option.

Wide resection is the main treatment for sarcomas; however, when they are located near major nerves, their sacrifices might affect limb function. The efficacy of ethanol adjuvant therapy for sarcomas has not been established. In this study, the anti-tumor effect of ethanol, as well as its neurotoxicity, were assessed. In vitro anti-tumor effect of ethanol as evaluated using MTT, wound healing, and invasion assays on a synovial sarcoma cell line (HS-SY-II). In vivo, an assessment was conducted in nude mice (implanted with subcutaneous HS-SY-II) treated with different ethanol concentrations after surgery with a close margin. Sciatic nerve neurotoxicity was assessed with electrophysiological and histological examination. In vitro, ethanol concentrations at 30% and higher showed cytotoxic effects in MTT assay and markedly reduced migration and invasive ability of HS-SY-II. In vivo, both 30% and 99.5% ethanol concentrations, compared to 0% concentration, significantly reduced the local recurrence. However, in the group treated with 99.5% ethanol, nerve conduction tests showed prolonged latency and decreased amplitude, and morphological changes suggestive of nerve degeneration were observed in the sciatic nerve, while the 30% ethanol did not cause neurological damage. In conclusion, 30% is the optimal concentration for ethanol adjuvant therapy after close-margin surgery for sarcoma.

Bone marrow aspirate (BMA) is an excellent source of cells and growth factors and has been used successfully for bone, cartilage, and soft-tissue healing. This study aimed to investigate the histological and biomechanical properties of autogenous tendon graft by injecting BMA and its protective effect against degenerative changes in a rabbit model of meniscal defects. Adult white rabbits were divided into untreated, tendon, and tendon + BMA groups, and meniscal defects were created in the knees. The tendon graft and articular cartilage status were evaluated by macroscopic and histological analysis at 4, 12, and 24 weeks postoperatively among the three groups. The tendon graft in the tendon and tendon + BMA groups were used for biomechanical evaluation at 4, 12, and 24 weeks postoperatively. The meniscal covering ratio in the tendon + BMA group was better than that in the tendon and untreated groups at 12 and 24 weeks postoperatively. The matrix around the central portion of cells in the tendon + BMA group was positively stained by safranin O and toluidine blue staining with metachromasia at 24 weeks. The histological score of the tendon graft in the tendon + BMA group was significantly higher than that in the untreated and tendon groups at 12 and 24 weeks postoperatively. In the tendon + BMA group, cartilage erosion was not shown at 4 weeks, developed slowly, and was better preserved at 12 and 24 weeks compared to the untreated and tendon groups. Histological scores for the articular cartilage were significantly better in the tendon + BMA group at 24 weeks. The compressive stress on the tendon graft in the tendon + BMA group was significantly higher than that in the tendon group at 12 weeks postoperatively. Transplantation of autogenous tendon grafts by injecting BMA improved the histologic score of the regenerated meniscal tissue and was more effective than the tendon and untreated group for preventing cartilage degeneration in a rabbit model of massive meniscal defects.