- 學位論文
低強度超音波刺激對於骨細胞之影響及其機轉討論
The Effects and Mechanism of Low-Intensity Ultrasound Stimulation on the Bone Cells
摘要
本研究主要探討低強度超音波之物理性刺激對於成骨細胞（osteoblasts）與蝕骨細胞（osteoclasts），以及坐骨神經切除（sciatic nerve section）與卵巢切除（ovariectomy）所造成骨質流失之影響。 超音波刺激對成骨細胞實驗中，是取用Sprague-Dawley（SD）新生鼠之頭蓋骨培養成骨細胞，在培養的第三天加入分化劑，隔天開始給予超音波刺激，連續刺激14天，其刺激參數為頻率1 MHz或45 kHz，給予連續波形或脈衝波形（on-time/off-time = 1：4），強度為50或120 mW/cm2下，每天刺激10分鐘。結果顯示超音波能促進骨小節形成和鹼性磷酸活性，及加強成骨細胞分化能力，尤其在刺激參數強度為120 mW/cm2，與PRF為10 3 Hz效果最佳（p < 0.05）。 超音波刺激對蝕骨細胞實驗中，蝕骨細胞是取用Sprague-Dawley（SD）公鼠之骨髓細胞，加入分化劑後，隔天開始給予超音波刺激，連續刺激6天，其刺激參數為：頻率1 MHz或45 kHz，脈衝波形on-time/off-time = 1：4，強度為120 mW/cm2，PRF分別為102及103 Hz下，每天刺激10分鐘。結果顯示，蝕骨細胞分化的形成數目減少（p < 0.01）。 超音波對動物骨質流失之實驗中，則是將Sprague-Dawley（SD）大鼠右側坐骨神經切除後，其刺激參數為：頻率45 kHz，連續波，強度為30 mW/cm2，每天刺激10分鐘，連續刺激4星期。在30 mW/cm2之超音波刺激下，對於坐骨神經切除骨質流失並無顯著影響。此外，對大鼠卵巢切除後骨質流失之實驗中，其刺激參數為：頻率1 MHz或45 kHz，連續波，強度分別為120 或30 mW/cm2，每天刺激10分鐘，連續刺激4星期。發現在超音波刺激下，對於卵巢切除的肢體側骨質流失也並無影響。
並列摘要
The purpose of this study was not only to examine the effect of low-intensity ultrasound physical stimulation on differentiation of osteoblasts and osteoclasts but also to investigate the effect of ultrasound stimulation on the bone loss induced by sciafic nerve section and ovariectomy. The effect of ultrasound stimulation on the maturation of osteoblast cell was examined. The osteoblasts were obtained from skulls of newborn Sprague-Dawley(SD) rats and cultured in ΚEM containing vitamin C and β-glycerophosphate, and the ultrasound stimulation was applied between Day-4 and Day-18, respectively. The parameters were: frequency = 1 MHz or 45 kHz, continuous or pulsed wave (on-time/off-time = 1：4), intensity = 50 or 120 mW/cm2, PRF = 10、102、103 or 104 Hz, and 10 minutes/day. It was found that ultrasound stimulation increased bone nodule numbers and alkaline phosphatase activity. The stimulation parameters were used: intensity = 120 mW/cm2 and PRF = 103 Hz. (p < 0.05) The effect of ultrasound stimulation on the formation of osteoclast was also investigated. The bone marrow stromal cells of Sprague-Dawley (SD) rats were cultured in DMEM containing M-CSF and RANKL to facilitate the differentiation of osteoclasts. Ultrasound stimulation was applied to cells between Day-2 and Day-7. Ultrasound parameters were: frequency = 1 MHz or 45 kHz, pulsed wave 1:4, intensity = 120 mW/cm2, PRF = 102 or 103 Hz, and 10 minutes/day. It was found that ultrasound stimulation inhibited the differentiation of osteoclasts. (p < 0.01) We examined the effect of ultrasound stimulation on boss loss induced by sciatic nerve section. Ultrasound parameters were: frequency = 45 kHz, continuous wave, intensity = 30 mW/cm2. It was applied to the joint of femur and tibia for 10 minutes per day (5 days /week and for 4 weeks). It was found that ultrasound stimulation exerts no significantly effect on the bone loss induced by the cut of sciatic nerve. In addition, the effect of ultrasound stimulation on boss loss induced by ovariectomy was also investigated. Ultrasound parameters were: frequency = 1 MHz or 45 kHz, continuous wave, intensity = 120 or 30 mW/cm2. It was applied to the joint of femur and tibia for 10 minutes per day (5 days /week and for 4 weeks). It was also found that ultrasound stimulation exerts no effect on the bone loss induced by ovariectomy.