Xiaolan He, Guangjie Bao, Linglu Sun, Xue Zhang, Shanying Bao, Hong Kang
Institute of Stomatology, Lanzhou University, Lanzhou 730000, China.
Institute of Stomatology, Lanzhou University, Lanzhou 730000, China;Key Laboratory of Oral Diseases of Gansu Provincial, Key Laboratory of Stomatology of State Ethnic Affairs Commission, Northwest Minzu University, Lanzhou 730030, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2017 Aug 1;35(4):362-367. doi: 10.7518/hxkq.2017.04.004.
Objective The effect of different oxygen tensions on the cytoskeleton remodeling of goat temporomandibular joint (TMJ) disc cells were investigated. Methods Goat TMJ disc cells were cultured under normoxia (21% O₂) and hypoxia (2%, 4%, and 8% O₂). Toluidine blue, picrosirius red, and type Ⅰ collagen immunocytochemical staining were performed to observe the changes in cell phenotype under different oxygen levels. Immunofluorescent staining and real-time reverse transcription-polymerase chain reaction analysis were then performed to identify actin, tubulin, and vimentin in the cultured disc cells. Results TMJ disc cells still displayed fibroblast characteristics under different oxygen levels and their cytoskeletons had regular arrangement. The fluorescence intensities of actin and vimentin were lowest at 4% O₂(P<0.05), whereas that of tubulin was highest at 2% O₂ (P<0.05). No significant difference among the other groups was observed (P>0.05). Actin mRNA levels were considerably decreased at 2% O₂ and 4% O₂ in hypoxic conditions, while actin mRNA expression was highest in 21% O₂. Tubulin mRNA levels considerably increased at 2% O₂, while tubulin mRNA expression was lowest in 8% O₂ (P<0.05). Vimentin mRNA expression was lowest at 4% O₂ and highest at 21% O₂, and significant differences were observed between vimentin mRNA expression levels among these oxygen levels (P<0.05). Conclusion Cytoskeletons were reconstructed in different oxygen tensions, and 2% O₂ may be the optimal oxygen level required to proliferate TMJ disc cells.
目的 研究不同氧张力对山羊颞下颌关节(TMJ)盘细胞细胞骨架重塑的影响。方法 将山羊TMJ盘细胞分别在常氧(21% O₂)和低氧(2%、4%和8% O₂)条件下培养。进行甲苯胺蓝、天狼星红和Ⅰ型胶原免疫细胞化学染色,观察不同氧水平下细胞表型的变化。然后进行免疫荧光染色和实时逆转录 - 聚合酶链反应分析,以鉴定培养的盘细胞中的肌动蛋白、微管蛋白和波形蛋白。结果 在不同氧水平下,TMJ盘细胞仍表现出成纤维细胞特征,其细胞骨架排列规则。肌动蛋白和波形蛋白的荧光强度在4% O₂时最低(P<0.05),而微管蛋白的荧光强度在2% O₂时最高(P<0.05)。其他组之间未观察到显著差异(P>0.05)。在低氧条件下,2% O₂和4% O₂时肌动蛋白mRNA水平显著降低,而在21% O₂时肌动蛋白mRNA表达最高。微管蛋白mRNA水平在2% O₂时显著增加,而在8% O₂时微管蛋白mRNA表达最低(P<0.05)。波形蛋白mRNA表达在4% O₂时最低,在21% O₂时最高,这些氧水平之间的波形蛋白mRNA表达水平存在显著差异(P<0.05)。结论 在不同氧张力下细胞骨架发生重构,2% O₂可能是TMJ盘细胞增殖所需的最佳氧水平。