低能体外冲击波和低剂量间歇甲状旁腺素干预成骨细胞的增殖和分化（英文）

Low-density extracorporeal shock wave and low-dose intermittent recombinant human parathyroid hormone 1-34 influence proliferation and differentiation of osteoblasts

背景:体外冲击波等应力刺激可促进成骨,甲状旁腺激素激素也参与调控骨代谢。目的:实验探讨低剂量间歇人重组甲状旁腺素1-34和低能体外冲击波对体外培养大鼠成骨细胞增殖及成骨分化的作用。方法:采用改良胶原酶消化法培养大鼠乳鼠颅骨来源成骨细胞备用。分别用60-150次0.18 mJ/mm2低能体外冲击波刺激体外培养大鼠成骨细胞,不同浓度(10-12 mol/L-10-10 mol/L)及作用方式的人重组甲状旁腺素1-34刺激,以及低能体外冲击波和间歇低剂量(10-11 mol/L)间歇人重组甲状旁腺素1-34刺激共同作用后,用锥虫蓝法进行细胞计数、MTT和流式细胞术分析检测大鼠成骨细胞的增殖情况;用酶标仪检测碱性磷酸酶活性,用免疫组化检测Ⅰ型胶原表达来观察大鼠成骨细胞的成骨分化。结果与结论:60-150次0.18 mJ/mm2低能体外冲击波刺激、间歇人重组甲状旁腺素1-34(10-11和10-10 mol/L)刺激以及低能体外冲击波+间歇人重组甲状旁腺素1-34(10-11 mol/L)刺激均可显著促进体外培养大鼠成骨细胞增殖和成骨分化(P<0.05),其中60-150次低能体外冲击波刺激+间歇人重组甲状旁腺素1-34刺激各组作用最强(P<0.05)。结果证实,适当的低能体外冲击波应力刺激和低剂量间歇人重组甲状旁腺素1-34刺激联合应用可显著促进体外培养大鼠成骨细胞的增殖和成骨分化。

BACKGROUND：Suitable stress signals such as extracorporeal shock wave （ESW） and parathyroid hormone can be involved in regulation of bone formation and metabolism.OBJECTIVE：To investigate the effects of low-density ESW and low-dose intermittent recombinant human parathyroid hormone 1-34 （rhPTH1-34） on the proliferation and differentiation of osteoblasts from rats cultured in vitro.METHODS：Osteoblasts from the skul of neonatal rats were cultured using col agenase digestion method. After 60-150 times of stimulations of 0.18 mJ/mm2 ESW, different patterns of rhPTH1-34 at 10-12-10-10 mol/L and ESW＋intermittent rhPTH1-34 （10-11 mol/L）, osteoblasts were col ected to observe cel proliferation by cel counting, MTT and flow cytometry analysis and to observe cel differentiation by measuring alkaline phosphatase and type I col agen expression.RESULTS AND CONCLUSION：0.18 mJ/mm 2 ESW （60-150 times）, intermittent rhPTH1-34 （10-11 and 10-10 mol/L） and ESW＋intermittent rhPTH1-34 （10-11 mol/L） stimulations could al improve proliferation and differentiation of rat osteoblasts cultured in vitro （P〈0.05）. ESW （60-150 times）＋intermittent rhPTH1-34 stimulation showed better effects than ESW and intermittent rhPTH1-34 alone （P〈0.05）. These findings indicate that the combination of suitable ESW stimulation and low-dose intermittent rhPTH1-34 stimulation can improve the proliferation and differentiation of rat osteoblasts significantly.