週期性壓力刺激以及其引發之細胞外基質金屬蛋白-3對蝕骨細胞的影響

在矯正治療的過程中，是藉由矯正裝置來對牙齒進行施力，而牙周組織，包含牙周韌帶，齒槽骨等，扮演著接受力量刺激的角色，並且產生一連串的代謝反應而使得牙齒移動。過去已知骨母細胞接受力學刺激後會調控蝕骨細胞的分化以進行骨質重塑活動，其中基質金屬蛋白酶-3(MMP-3可能扮演重要角色。同時，蝕骨細胞本身亦可能是直接接受力學刺激的細胞，而力量刺激可能會影響蝕骨細胞的分化。本研究以小鼠RAW 264.7細胞為對象，培養8~132小時，觀察MMP-3及RANKL對於蝕骨細胞分化的影響。以即時定量聚合酶鏈鎖反應(Real-time PCR)分析，結果在同時加入RANKL及MMP-3的細胞其TRAP及MMP-9基因表現量會較僅加入RANKL為多，並且在染色的結果上會有較多TRAP染色的細胞，同時也會有較多鈣化物質吸收之能力。這證明MMP-3與RANKL對於蝕骨細胞的分化調控具有加成作用。另外並將RAW264.7細胞培養於3D膠原蛋白凝集體中，施予細胞1%週期性壓力刺激 24小時，並給予有無RANKL刺激。而蝕骨細胞在直接接受週期性壓力後，會有形態上的改變，但是其TRAP表現上並無顯著差異。證明蝕骨細胞亦會受到力學刺激調控，但並非使其成熟分化。

關鍵字

基質金屬蛋白酶機械力量刺激； RAW 264.7

並列摘要

Orthodontic tooth movement is based on the force delivered by the appliance and transducing to the surrounding tissue such as periodontal ligament , and alveolar bone to recieve the mechanical stimulation. The serial metabolic effect after force stimulation that known as bone remodeling involves both osteoblast and osteoclast. Osteoblast can regulate osteoclast function after force stimulation, and MMP-3 is proposed to be one of the key factors. Besides, osteoclast might be stimulated by mechanical force directly. We used mouse monocyte cell line RAW 264.7 cell, cultured with RANKL and MMP-3 to evaluate the indirect mechanical stimulation on osteoclast. In order to verify how the mechanical compression may affect osteoclast directly, 1% cyclic compression force for 24 hours with or without RANKL was applied to RAW 264.7 cells grown in a 3D collagen gel to mimic the physiologic environment. The result of real-time PCR showed that MMP-9 and TRAP genes were up regulated significantly after 84 hours cultured with combination of RANKL and MMP-3 then with RANKL only. TRAP stain and pit assay indicated more mature osteoclast function in the combination group. Therefore, we suggest a synergic effect of MMP-3 and RANKL in regulating osteoclast differentiation. Force stimulation also cause morphological change of osteoclast without significant change of TRAP expression, which suggest osteoclastmay have mechanical receptor itself, but not this direct influence the differentiation process.

並列關鍵字

matrix metalloproteinase ； mechanical stress ； RAW 264.7 osteoclast

參考文獻

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週期性壓力刺激以及其引發之細胞外基質金屬蛋白-3對蝕骨細胞的影響

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