This study was carried out to investigate the protective effect of prednisolone in rabbit primary cultured articular chondrocytes treated with sodium nitroprusside (SNP), a nitric oxide donor. After a cell phenotype was determined, the MTT assay and Western blot analysis of type II collagen, cylooxygenase-2 (COX-2) and phosphorylated extracellular regulated kinase (pERK) were performed in the control, SNP (298 μg/ml) alone or SNP plus prednisolone (0.05-50 μg/ml)-treated rabbit articular chondrocytes. Immunofluorescence staining of type II collagen was also performed. Cell morphology indicated that SNP treatment induced cytotoxicity, and that the SNP-induced cytotoxicity was inhibited by prednisolone treatment. MTT assay showed that the SNP treatment resulted in a significant decrease in the level of cell viability compared with that of control (p<0.01), and that the prednisolone treatment resulted in a decrease in the SNP-induced cytotoxicity. SNP treatment resulted in a decrease in the level of type II collagen, compared with the control chondrocytes. The prednisolone treatment recovered the down-regulated expression of type II collagen induced by SNP, showing a significant level in 5 μg/ml of the prednisolone treatment group compared to the SNP treatment group (p<0.05). A significant increase in COX-2 was significantly induced by the SNP treatment compared to control chondrocytes (p<0.01). The COX-2 expression was decreased by the prednisolone treatment, showing a significant level in 50 μg/ml of the prednisolone treatment group compared to the SNP treatment group (p<0.05). These phenomena was confirmed by immunofluorescence staining. Furthermore, the SNP treatment significantly induced a decrease of pERK expression compared to the control chondrocytes (p<0.01). The prednisolone treatment recovered its expression, showing a significant level in 0.5 μg/ml of the prednisolone treatment group compared to the SNP treatment group (p<0.05). Taken the above results together, prednisolone is considered to inhibit SNP-induced cell death and dedifferentiation, and modulated expression of COX-2 and pERK in rabbit articular chondrocytes.

Recent experimental researches supports the hypothesis that mechanical stimuli play a role in regulating specialized molecules expression in articular cartilage in vitro and in vivo. Therefore we are studying bioreactor to provide a stimulus of mimicked real knee environment. Our bioreactor system consists of a joint motion stimulation device and culture chamber. We fabricated a stimulation system similar to movement of the CPM(Continuous Passive Motion) device for human joint motion. This system is composed of a linear stage, ball spline and links. Also we fabricated a culture system for media and circulation and mimicked knee environment. However, Previous culture system has some problems that media leakage and interference of joint model. In this study, we improved a new-chamber system for non-interference of joint model and medium sealing. Then, this system could be used to investigate the effect of dynamic culture.

본 연구는 관절통 모델에서 웅담 우황과 웅담 우황 사향의 약침액의 효과를 검사하기 위해 수행되었다. 할로탄 마취하에서 관절통은 수컷 쥐의 관절강내에 2% carrageenan을 주입하여 유발시켰다. 운동자극에 대한 감각신경의 반응은 약침을 시술하기 전과 후에 기록하였다. 경혈에 주입한 약침은 유해한 운동 자극에 대한 신경의 반응을 억제시켰다 족삼리에 시술한 약침은 합곡에 비해 유해한 자극에 대한 관절 감각신경의 반응을 더 많이 억제시켰다. 이러한 결과는 웅담 우황과 웅담 우황 사향의약침이 관절통을 완화하는 데 효과적인 치료법을 제공할 수 있다는 것을 시사한다.