Yin Tsung-Cheng, Wang Ching-Jen, Yang Kunder D, Wang Feng-Sheng, Sun Yi-Chih
Department of Orthopedic Surgery, Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan.
Chang Gung Med J. 2011 Jul-Aug;34(4):367-74.
This in vitro study investigated the angiogenesis and osteogenesis effects of shockwaves on bone marrow stromal cells (BMSCs) from hips with osteonecrosis.
BMSCs were harvested from the bone marrow cavity of the proximal femur in six patients with osteonecrosis of the femoral head. The specimens were divided into four groups, the control, shockwave, shockwave plus nω-nitro- L-arginine methyl ester (L-NAME) and a nitric oxide (NO) donor (NOC18) groups. The control group received no shockwaves and was used as the baseline. The shockwave group received 250 shockwave impulses at 14 Kv (equivalent to 0.18 mJ/mm2 energy flux density). The shockwave plus LNAME group was pre-treated with L-NAME before receiving shockwaves. The NOC18 group received NOC18 after cell culture for 48 hours. The evaluations included cell proliferation (MTT) assay, alkaline phosphatase, real time reverse transcriptase-polymerase chain reaction analysis of vessel endothelial growth factor (VEGF), bone morphogenic protein (BMP)-2, RUNX2 and osteocalcin mRNA expression and von Kossa stain for mineralized nodules.
The shockwave group showed significant increases in MTT, VEGF, alkaline phosphatase, BMP2, RUNX2 and osteocalcin mRNA expression and more mature mineralized nodules compared with the control. Pre-treatment with L-NAME significantly reduced the angiogenic and osteogenic effects of extracorporeal shockwave therapy (ESWT) and the results were comparable with the control. Administration of NOC18 significantly enhanced the angiogenesis and osteogenesis effects compared with the control and the results were comparable with the shockwave group.
ESWT significantly enhanced the angiogenic and osteogenic effects of BMSCs mediated through the NO pathway in hips with osteonecrosis. These innovative findings, at least in part, explain some of the mechanism of shockwaves in osteonecrosis of the hip.
本体外研究探讨了冲击波对股骨头坏死患者髋部骨髓间充质干细胞(BMSCs)的血管生成和成骨作用。
从6例股骨头坏死患者的股骨近端骨髓腔中采集BMSCs。标本分为四组,即对照组、冲击波组、冲击波加Nω-硝基-L-精氨酸甲酯(L-NAME)组和一氧化氮(NO)供体(NOC18)组。对照组不接受冲击波,作为基线。冲击波组接受14千伏的250次冲击波脉冲(相当于0.18毫焦/平方毫米的能量通量密度)。冲击波加L-NAME组在接受冲击波前用L-NAME预处理。NOC18组在细胞培养48小时后接受NOC18。评估包括细胞增殖(MTT)测定、碱性磷酸酶、血管内皮生长因子(VEGF)、骨形态发生蛋白(BMP)-2、RUNX2和骨钙素mRNA表达的实时逆转录聚合酶链反应分析以及矿化结节的冯·科萨染色。
与对照组相比,冲击波组的MTT、VEGF、碱性磷酸酶、BMP2、RUNX2和骨钙素mRNA表达显著增加,矿化结节更成熟。用L-NAME预处理显著降低了体外冲击波疗法(ESWT)的血管生成和成骨作用,结果与对照组相当。与对照组相比,给予NOC18显著增强了血管生成和成骨作用,结果与冲击波组相当。
ESWT显著增强了股骨头坏死患者髋部BMSCs通过NO途径介导的血管生成和成骨作用。这些创新性发现至少部分解释了冲击波在髋部骨坏死中的一些作用机制。