Yao Meng, Man Liu, Ding Bai
Hua Xi Kou Qiang Yi Xue Za Zhi. 2015 Apr;33(2):130-4. doi: 10.7518/hxkq.2015.02.005.
To investigate the functions of human periodontal myofibroblast (MFB) in vitro.
Human periodontal fibroblast (hPDLFs) was cultured and induced to MFB by transforming growth factor-β1 (TGF-β1). MFB was denoted as the experimental group, whereas the hPDLFs was the control group. The groups were continuously cultured and harvested at 0, 12, 24, 48, and 72 h. The MFB marker α-smooth muscle actin (α-SMA) was examined by immunocytochemistry. The expression of fibronectin (FN) between MFB was examined by immunocytochemistry to detect the MFB contact relationship. The mRNA expression levels of α-SMA, collagen (Col) I, and Col III were measured by reverse transcription-polymerase chain reaction (RT-PCT) to analyze extracellular matrix secretion. The protein expression levels of α-SMA and Col I were also assessed by Western blot.
The experimental group had significantly higher α-SMA expression than the control group at 0 h (P < 0.001). A positive expression of FN was found between MFB. The experimental group had significantly higher expression levels of Col I and Col III than the control group at 24 h (P < 0.001).
Human periodontal MFB presents a continuous, high expression of α-SMA. MFB could interact through FN. MFB is significantly capable of extracellular matrix secretion.
体外研究人牙周肌成纤维细胞（MFB）的功能。
培养人牙周膜成纤维细胞（hPDLFs），并用转化生长因子-β1（TGF-β1）诱导其分化为MFB。将MFB作为实验组，hPDLFs作为对照组。两组细胞持续培养，并于0、12、24、48和72小时收获。通过免疫细胞化学检测MFB标志物α-平滑肌肌动蛋白（α-SMA）。通过免疫细胞化学检测MFB之间纤连蛋白（FN）的表达，以检测MFB的接触关系。通过逆转录-聚合酶链反应（RT-PCT）测量α-SMA、胶原蛋白（Col）I和Col III的mRNA表达水平，以分析细胞外基质分泌情况。还通过蛋白质免疫印迹法评估α-SMA和Col I的蛋白质表达水平。
实验组在0小时时α-SMA表达显著高于对照组（P < 0.001）。在MFB之间发现FN呈阳性表达。实验组在24小时时Col I和Col III的表达水平显著高于对照组（P < 0.001）。
人牙周MFB呈现α-SMA的持续高表达。MFB可通过FN相互作用。MFB具有显著的细胞外基质分泌能力。
