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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: Stress shock such as extracorporeal shock waves can promote osteogenesis, and parathyroid hormone is also involved in the regulation of bone metabolism. OBJECTIVE: To investigate the effect of low-dose intermittent recombinant human parathyroid hormone 1-34 and low-energy extracorporeal shock wave on the proliferation and osteogenic differentiation of cultured rat osteoblasts. Methods: Rat calvarial osteoblasts were prepared by a modified collagenase digestion method. The cultured rat osteoblasts were stimulated with 60-150 low-energy extracorporeal shock waves of 0.18 mJ / mm2 respectively, and the concentrations of human recombinant parathyroid hormone 1 (10-12 mol / L-10-10 mol / L) and the mode of action -34 stimulation, combined with low-energy extracorporeal shock waves and intermittent low-dose (10-11 mol / L) intermittent human recombinant parathyroid hormone 1-34 stimulation, cell counting was performed using the trypan blue method, MTT and flow cytometry The proliferation of rat osteoblasts was analyzed. The alkaline phosphatase activity was detected by microplate reader. The expression of collagen Ⅰ was detected by immunohistochemistry to observe the osteoblastic differentiation of rat osteoblasts. RESULTS AND CONCLUSION: 60-150 low-energy extracorporeal shock waves of 0.18 mJ / mm2, intermittent human recombinant parathyroid 1-34 (10-11 and 10-10 mol / L) stimulation and low energy extracorporeal shock wave + intermittent human recombinant parathyroid The stimulation with 10-11 mol / L of noradrenaline could significantly promote the proliferation and osteogenic differentiation of rat osteoblasts cultured in vitro (P <0.05), of which 60-150 low-energy extracorporeal shock waves + intermittent human recombinant thyroid Psoriatic 1-34 stimulated the strongest effect in each group (P <0.05). The results show that appropriate low-ESB stress stimulation and intermittent low-dose intermittent recombinant human parathyroid hormone 1-34 stimulation can significantly promote in vitro cultured rat osteoblast proliferation and osteogenic differentiation.