Huang Linjian, Cai Xieyi, Li Hui, Xie Qianyang, Zhang Min, Yang Chi
Department of Oral and Maxillofacial Surgery, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai 200011, China; Department of Oral and Maxillofacial Surgery, The Second Affiliated Hospital Zhejiang University School of Medicine, Hangzhou 310009, China.
Department of Oral and Maxillofacial Surgery, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai 200011, China.
Arch Oral Biol. 2015 Apr;60(4):622-30. doi: 10.1016/j.archoralbio.2015.01.003. Epub 2015 Jan 13.
The goal of the study was to investigate the production of collagen, type II, alpha 1 (COL2A1), SOX9, alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), Indian hedgehog (Ihh), and periarticular cell-derived parathyroid hormone-related protein (PTHrP) in mandibular condylar chondrocytes under static pressure stimuli.
Mandibular condylar chondrocytes separated from rabbit temporomandibular joint (TMJ) were treated with a static pressure of 100kPa for 0, 1, 2, 3, and 4h by an in-house-designed pressure chamber. A CCK-8 kit was used to analyze the cell viability. The production of COL2A1, SOX9, ALP, Runx2, Ihh, and PTHrP was detected by Western blot or real-time polymerase chain reaction (PCR). Changes in cell morphology were observed by scanning electron microscopy.
Compared with the control group (0h), the cytoplasmic processes of treated chondrocytes obviously increased and elongated, and the cell viability of pressurized chondrocytes were 91.13% (1h), 103.41% (2h), 103.47% (3h), and 104.94% (4h), respectively. The exposure of condylar chondrocytes to a static pressure of 100kPa for 3-4h resulted in a significant increase in COL2A1, SOX9, ALP, and Runx2. After a static pressure loading of 100kPa, the activation of Ihh and PTHrP was also observed.
Mandibular condylar chondrocytes adapt to alterations of the microenvironment. Ihh and PTHrP are sensitive to static pressure. Our findings suggest that static pressure accelerated the chondrogenic and osteogenic differentiation of condylar chondrocytes, which may influence the pathological progress of temporomandibular diseases.
本研究旨在探讨静压力刺激下下颌髁突软骨细胞中Ⅱ型胶原α1(COL2A1)、SOX9、碱性磷酸酶(ALP)、 runt相关转录因子2(Runx2)、印度刺猬因子(Ihh)及关节周围细胞源性甲状旁腺激素相关蛋白(PTHrP)的产生情况。
采用自行设计的压力室,对从兔颞下颌关节(TMJ)分离出的下颌髁突软骨细胞施加100kPa的静压力,分别作用0、1、2、3和4小时。使用CCK-8试剂盒分析细胞活力。通过蛋白质免疫印迹法或实时聚合酶链反应(PCR)检测COL2A1、SOX9、ALP、Runx2、Ihh和PTHrP的产生情况。通过扫描电子显微镜观察细胞形态变化。
与对照组(0小时)相比,处理后的软骨细胞胞质突起明显增多、变长,加压处理的软骨细胞活力分别为91.13%(1小时)、103.41%(2小时)、103.47%(3小时)和104.94%(4小时)。将髁突软骨细胞暴露于100kPa的静压力下3 - 4小时,导致COL2A1、SOX9、ALP和Runx2显著增加。在施加100kPa静压力后,还观察到Ihh和PTHrP的激活。
下颌髁突软骨细胞能适应微环境的改变。Ihh和PTHrP对静压力敏感。我们的研究结果表明,静压力加速了髁突软骨细胞的软骨形成和成骨分化,这可能影响颞下颌疾病的病理进程。