Mechanical Stimulus Inhibits the Growth of a Bone Tissue Model Cultured In Vitro

Zong-ming Wan; Lu Liu; Jian-yu Li; Rui-xin Li; Yong Guo; Hao Li; Jian-ming Zhang; Xi-zheng Zhang

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Mechanical Stimulus Inhibits the Growth of a Bone Tissue Model Cultured In Vitro

机译：机械刺激抑制体外培养的骨组织模型的生长

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Objectives To construct the cancellous bone explant model and a method of culturing these bone tissues in vitro, and to investigate the effect of mechanical load on growth of cancellous bone tissue in vitro. Methods Cancellous bone were extracted from rabbit femoral head and cut into 1-mm-thick and 8-mm-diameter slices under sterile conditions. HE staining and scanning electron microscopy were employed to identify the histomorphology of the model after being cultured with a new dynamic load and circulating perfusion bioreactor system for 0, 3, 5, and 7 days, respectively. We built a three-dimensional model using microCT and analyzed the loading effects using finite element analysis. The model was subjected to mechanical load of 1000, 2000, 3000, and 4000μεrespectively for 30 minutes per day. After 5 days of continuous stimuli, the activities of alkaline phosphatase (AKP) and tartrate-resistant acid phosphatase (TRAP) were detected. Apoptosis was analyzed by DNA ladder detection and caspase-3/8/9 activity detection. Results After being cultured for 3, 5, and 7 days, the bone explant model grew well. HE staining showed the apparent nucleus in cells at the each indicated time, and electron microscope revealed the living cells in the bone tissue. The activities of AKP and TRAP in the bone explant model under mechanical load of 3000 and 4000μεwere significantly lower than those in the unstressed bone tissues (all P<0.05). DNA ladders were seen in the bone tissue under 3000 and 4000μεmechanical load. Moreover, there was significant enhancement in the activities of caspase-3/8/9 in the mechanical stress group of 3000 and 4000με(all P<0.05). Conclusions The cancellous bone explant model extracted from the rabbit femoral head could be alive at least for 7 days in the dynamic load and circulating perfusion bioreactor system, however, pathological mechanical load could affect the bone tissue growth by apoptosis in vitro. The differentiation of osteoblasts and osteoclasts might be inhibited after the model is stimulated by mechanical load of 3000 and 4000με.

机译：目的建立松质骨外植体模型及体外培养这些骨组织的方法，探讨机械负荷对松质骨组织生长的影响。方法从兔股骨头中提取松质骨，并在无菌条件下切成1毫米厚和8毫米直径的切片。 HE染色和扫描电镜观察分别用新的动态负荷和循环灌注生物反应器系统培养0、3、5和7天后，鉴定模型的组织形态。我们使用microCT建立了三维模型，并使用有限元分析法分析了加载效果。每天分别对模型施加1000、2000、3000和4000με的机械负载，持续30分钟。连续刺激5天后，检测到碱性磷酸酶（AKP）和抗酒石酸酸性磷酸酶（TRAP）的活性。通过DNA梯检测和caspase-3 / 8/9活性检测分析细胞凋亡。结果培养3、5和7天后，植骨模型生长良好。 HE染色显示在每个指示的时间在细胞中有明显的细胞核，而电子显微镜显示出骨组织中的活细胞。在3000和4000με的机械载荷下，骨外植体模型中AKP和TRAP的活性明显低于未受压的骨组织中的AKP和TRAP的活性（均P <0.05）。在3000和4000με的机械载荷下，在骨组织中观察到DNA梯。此外，在3000和4000με的机械应力组中，caspase-3 / 8/9的活性显着增强（均P <0.05）。结论从兔股骨头中提取的松质骨外植体模型在动态负荷和循环灌注生物反应器系统中至少可以存活7天，但是病理机械负荷可能通过体外细胞凋亡影响骨组织的生长。在3000和4000με的机械负荷刺激模型后，可能抑制成骨细胞和破骨细胞的分化。

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《中国医学科学杂志（英文版）》 |2013年第4期|218-224|共7页
作者
Zong-ming Wan; Lu Liu; Jian-yu Li; Rui-xin Li; Yong Guo; Hao Li; Jian-ming Zhang; Xi-zheng Zhang;
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Institute of Medical Equipment, Academy of Military Medical Sciences of Chinese People's Liberation Army, Tianjin 300161, China;

Institute of Medical Equipment, Academy of Military Medical Sciences of Chinese People's Liberation Army, Tianjin 300161, China;

Institute of Medical Equipment, Academy of Military Medical Sciences of Chinese People's Liberation Army, Tianjin 300161, China;

Institute of Medical Equipment, Academy of Military Medical Sciences of Chinese People's Liberation Army, Tianjin 300161, China;

Institute of Medical Equipment, Academy of Military Medical Sciences of Chinese People's Liberation Army, Tianjin 300161, China;

Institute of Medical Equipment, Academy of Military Medical Sciences of Chinese People's Liberation Army, Tianjin 300161, China;

Institute of Medical Equipment, Academy of Military Medical Sciences of Chinese People's Liberation Army, Tianjin 300161, China;

Institute of Medical Equipment, Academy of Military Medical Sciences of Chinese People's Liberation Army, Tianjin 300161, China;

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收录信息中国科学引文数据库(CSCD);中国科技论文与引文数据库(CSTPCD);
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Mechanical Stimulus Inhibits the Growth of a Bone Tissue Model Cultured In Vitro

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