周期性压应力诱导髁突软骨细胞分泌促破骨因子的研究
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摘要
目的:研究周期性压应力对髁突软骨细胞促破骨因子表达的影响。方法:分离原代大鼠髁突软骨细胞,并与琼脂糖培养基混合,采用Flexcell细胞加力系统对软骨细胞/琼脂糖混合物进行压应力加载,分别对细胞施加15、30及45 kPa周期性压应力(频率为0.5 Hz),加力时间分别为2、6及12 h。对照组采用相同处理方式,但不施加压应力。加力结束后收集细胞,采用实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,Real-time PCR)及Western blot检测软骨细胞促破骨因子表达情况。结果:15 kPa压力刺激软骨细胞6 h即可促进软骨细胞核因子-κB(nuclear factor kappa-B, NF-κB)配体受体激活剂(the receptor activator of NF-κB ligand,RANKL)的表达(P<0.05),30及45 kPa压力刺激软骨细胞6及12 h可促进软骨细胞基质衍生因子1(stroma derived factor 1,SDF-1)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)及RANKL的表达(P<0.05),但其单核细胞趋化蛋白-1(monocyte chemotactic protein-1,MCP-1)、巨噬细胞集落刺激因子(macrophage-colony stimulating factor,M-CSF)及骨保护素(osteoprotegerin,OPG)的表达则与对照组比较差异无统计学意义(P>0.05)。结论:周期性压应力可促进软骨细胞表达SDF-1、TGF-β1及RNAKL等促破骨因子。
Objective:To investigate the effects of periodic compression on the expression of pro-osteoclastic factors by mandibular condylar chondrocytes(MCCs). Methods:The rat MCCs were isolated and mixed with agarose culture medium. The Flexcell system was used to conduct compression on the MCCs/agarose mixture. The intensities of compression were 15, 30, and 45 kPa(frequency 0.5 Hz), and the stimulating time were 2, 6, and 12 hours. The control group was treated in the same way but without compression. Cells were collected after the application of force and the expression of pro-osteoclastic factors by MCCs were detected. Results:When the MCCs were stimulated by 15 kPa compression for 6 hours, the expression of RANKL was significantly higher than those of the controls. When the MCCs were stimulated by 30 and 45 kPa compression for 6 and 12 hours, the expression of SDF-1, TGF-beta-1, and RANKL were higher than those of the controls, but the gene expression of MCP-1, M-CSF, and OPG were not different from those of the controls. Conclusion: Periodic compression could promote the expression of pro-osteoclastic factors such as SDF-1, TGF-beta 1, and RANKL.
Objective:To investigate the effects of periodic compression on the expression of pro-osteoclastic factors by mandibular condylar chondrocytes(MCCs). Methods:The rat MCCs were isolated and mixed with agarose culture medium. The Flexcell system was used to conduct compression on the MCCs/agarose mixture. The intensities of compression were 15, 30, and 45 kPa(frequency 0.5 Hz), and the stimulating time were 2, 6, and 12 hours. The control group was treated in the same way but without compression. Cells were collected after the application of force and the expression of pro-osteoclastic factors by MCCs were detected. Results:When the MCCs were stimulated by 15 kPa compression for 6 hours, the expression of RANKL was significantly higher than those of the controls. When the MCCs were stimulated by 30 and 45 kPa compression for 6 and 12 hours, the expression of SDF-1, TGF-beta-1, and RANKL were higher than those of the controls, but the gene expression of MCP-1, M-CSF, and OPG were not different from those of the controls. Conclusion: Periodic compression could promote the expression of pro-osteoclastic factors such as SDF-1, TGF-beta 1, and RANKL.
引文
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[11] Campbell IK, Ianches G, Hamilton JA. Production of macrophage colony-stimulating factor (M-CSF) by human articular cartilageand chondrocytes. Modulation by interleukin-1 and tumor necrosis factor alpha [J]. Biochim Biophys Acta, 1993, 1182(1):57-63.
[12] Lin PM, Chen CT, Torzilli PA. Increased stromelysin-1(MMP-3) proteoglycan degradation(3B-and 7D4)and collagen damage in cyclically load-injured articular cartilage [J]. Osteoarthritis Cartilage, 2004, 12(6):485-496.
[2] Amarasekara DS, Yun H, Kim S, et al. Regulation of osteoclast differentiation by cytokine networks [J]. Immune Netw, 2018, 18(1):e8.
[3] Ono T, Nakashima T. Recent advances in osteoclast biology [J]. Histochem Cell Biol, 2018, 149(4):325-341.
[4] Betti BF, Everts V, Ket JCF, et al. Effect of mechanical loading on the metabolic activity of cells in the temporomandibular joint: a systematic review [J]. Clin Oral Investig, 2018, 22(1):57-67.
[5] 蔡长安, 张丽. 咬合干扰对大鼠髁突软骨细胞内质网应激及凋亡的影响[J].口腔医学研究,2015,31(1):23-26.
[6] Pérez Del Palomar A, Doblaré M. Finite element analysis of the temporomandibular joint during lateral excursions of the mandible [J]. J Biomech, 2006, 39(12):2153-2163.
[7] Miyata S, Furukawa K, Ushid T, et al. Static and dynamic mechanical properties of extracellular matrix synthesized by cultured chondrocytes [J]. Mater Sci Eng C Mater Biol Appl, 2004, 24(3):425-429.
[8] Utreja A, Yadav S, Villa MM, et al. Cell and matrix response of temporomandibular cartilage to mechanical loading [J]. Osteoarthritis Cartilage, 2016, 24(2):335-344.
[9] Usui M, Xing L, Drissi H, et al. Murine and chicken chondrocytes regulate osteoclastogenesis by producing RANKL in response to BMP2 [J]. J Bone Miner Res, 2008, 23(3):314-325.
[10] Masuyama R, Stockmans I, Torrekens S, et al. Vitamin D receptor in chondrocytes promotes osteoclastogenesis and regulates FGF23 production in osteoblasts [J]. J Clin Invest, 2006, 116(12):3150-3159.
[11] Campbell IK, Ianches G, Hamilton JA. Production of macrophage colony-stimulating factor (M-CSF) by human articular cartilageand chondrocytes. Modulation by interleukin-1 and tumor necrosis factor alpha [J]. Biochim Biophys Acta, 1993, 1182(1):57-63.
[12] Lin PM, Chen CT, Torzilli PA. Increased stromelysin-1(MMP-3) proteoglycan degradation(3B-and 7D4)and collagen damage in cyclically load-injured articular cartilage [J]. Osteoarthritis Cartilage, 2004, 12(6):485-496.