摘要
目的:从细胞水平探讨低强度脉冲超声波对体外分离培养的人骨膜细胞的生物学效应,分析低强度脉冲超声波可能的作用途径,为合理化的临床应用提供实验依据。方法:实验于2002-05/11在香港中文大学矫形外科学系完成。选取香港中文大学威尔士亲王医院矫形外科收治的1例27岁尺骨骨不连患者,以其尺骨骨膜培养的传2代细胞用于实验。按低强度脉冲超声波(30mW/cm2)作用于体外培养的人骨膜细胞的时间剂量分为4组:5min/d组,10min/d组,20min/d组,空白对照组。在超声波作用1,2,4d后检测各组DNA合成量、四甲基偶氮唑蓝摄取情况、碱性磷酸酶活性及骨钙素分泌量。结果:与空白对照组比较,5min/d组,10min/d组,20min/d组给予低强度脉冲超声波后,骨膜细胞DNA合成及四甲基偶氮唑蓝摄取量无明显变化(P>0.05);10min/d组与20min/d组骨膜细胞单位质量蛋白中碱性磷酸酶活性显著升高(P<0.05);20min/d组骨膜细胞受1,25-二羟基维生素D3刺激后分泌骨钙素显著增加,形成钙结节能力显著增强(P<0.01)。结论:20min/d以内剂量的低强度脉冲超声波对人骨膜细胞的生长增殖无明显影响,但能够增强人骨膜细胞碱性磷酸酶活性的表达,促进细胞分泌骨钙素及钙盐沉积,具有显著促进人骨膜细胞向成骨细胞分化的生物学作用。
AIM： To investigate the biological effect of low intensive pulsed ultrasound wave （LIPUS） on human periosteal cells in vitro from cell level, and analyze the possible action pathway of LIPUS wave, so as to provide experimental evidence for the reasonable clinical application. METHODS： The experiment was carried out in the Department of Orthopaedics, Chinese University of Hong Kong between May and November 2002. A 27-year-old patient with ulnar nonunion was selected from the Department of Orthopaedics, Prince of Wales Hospital, Chinese University of Hong Kong, the cells of 2 generations cultured in ulnar periost were used in the experiment. According to the time doses of LIPUS （30 mW/cm^2） acted on the in vitro cultured human periosteal cells, the cells were divided into with dose of 5, 10 and 20 minutes per day groups and blank control group. At 1, 2 and 4 days, the amount of DNA synthesis, thymidine incorporation of methyl-thiazol-tetrazolium （MTT）, activity of alkaline phosphatase and amount of osteocalcin secretion were detected. RESULTS： As compared with the blank control group, the DNA synthesis of periosteal cells and thymidine incorporation of MTT had no obvious changes after LIPUS treatment in the 5, 10 and 20 minutes per day groups, the activities of alkaline phosphatase in protein of unit mass in periosteal cells were significantly increased in the 10 and 20 minutes per day groups （P 〈 0.05）. In the 20 minutes per day group, after stimulated by 1,25-（OH） 2-vitamin D3, the osteocalcin secretion was dramatically increased, and the ability in forming calcium nod was significantly enhanced. （P 〈 0.01）. CONCLUSION： The results suggested that the LIPUS with the dosage within 20 minutes per day had no obvious influence on the growth and proliferation of human periosteal cells, but could enhance the expression of alkaline phosphatase activity in human periosteal cells, promote the osteocalcin secretion and deposition of calcium salts, it had the significant biological effect in promoting human periosteal cells to differentiate into osteoblasts.
出处
《中国临床康复》
CSCD
北大核心
2006年第5期78-80,共3页
Chinese Journal of Clinical Rehabilitation
基金
香港研究基金会项目(CUHK4257/99M)~~
