矽酸鈣材料結合纖維母細胞生長因子促進硬組織修復協同效應探討

骨頭修復的過程中需要周圍環境的生長因子的相互作用以來促進骨頭的生成。本研究主要探討矽酸鈣（calcium silicate，CS）和三鈣磷酸鹽（tricalcium phosphate，β-TCP）骨水泥經由成纖維母細胞生長因子受體（fibroblast growth factor receptor，FGFR）和MAPK路徑影響人類牙髓細胞（human dental pulp cells，hDPCs）的細胞行為。hDPCs培養於β-TCP和CS後，再使用PrestoBlue®和西方墨點法來分析細胞的生長及牙源性分化蛋白。此外還使用小干擾RNA（siRNA）轉染來抑制FGFR的表現。結果顯示CS除了促進hDPCs細胞的生長和FGFR表現外，也還會增強ERK和p38的蛋白活性。此外，在si-FGFR轉染後的hDPCs細胞培養於CS會抑制ERK的表現以及鈣沉積，這些結果都可證明ERK/MAPK路徑參與了矽離子影響hDPCs的牙源性分化。目前在臨床上，三氧礦化合物（MTA）是一常見的矽酸鈣材料，並且擁有了極佳的生物相容性。因此後續也嘗試將MTA/第二型纖維細胞生長因子-2（fibroblast growth factor-2，FGF-2）混合，並評估其物化性質以及對於hDPCs的影響。從結果得知，添加FGF-2的MTA並不會降低其硬化時間及強度，此外MTA/FGF-2還可以有效提升hDPCs細胞生長和分化能力。經過FGFR-siRNA轉染後，hDPCs細胞培養於有無FGF-2的MTA組別的ALP和OC的蛋白分泌則無顯著差異。這些結果顯示，CS會藉由通過FGFR來調節hDPCs細胞中ERK/MAPK活化，並且促進牙本齒質形成的關鍵作用。在臨床的應用上也可將MTA添加入FGF-2以提高hDPCs細胞的生長和牙本質分化的能力。

關鍵字

人類牙髓細胞；矽酸鈣；三氧礦化合物；纖維母細胞生長因子受體；有絲分裂活化蛋白激

並列摘要

Bone healing needs a complex interaction of growth factors that establishes an environment for efficient bone formation. We examine how calcium silicate (CS) and tricalcium phosphate (β-TCP) cements influence the behavior of human dental pulp cells (hDPCs) through fibroblast growth factor receptor (FGFR) and active MAPK pathways, in particular ERK. The hDPCs are cultured with β-TCP and CS, after which the cells’ viability and odontogenic differentiation markers are determined by using PrestoBlue® assay and western blot, respectively. The effect of small interfering RNA (siRNA) transfection targeting FGFR was also evaluated. The results showed CS promoted cell proliferation and enhances FGFR expression. It was also found that CS increases ERK and p38 activity in hDPCs. Furthermore, raises the expression and secretion of DSP, and DMP-1. Additionally, statistically significant differences (p < 0.05) have been found in the calcium deposition in si-FGFR transfection and ERK inhibitor between CS and β-TCP; these variations are indicated that ERK/MAPK signaling is involved in the silicon-induced odontogenic differentiation of hDPCs. In addition, Mineral trioxide aggregate (MTA) is a biocompatible material and has had several clinical applications in endodontic treatment since 1993. In this study, we examined the effects of MTA/fibroblast growth factor-2 (FGF-2) on material properties and in vitro hDPCs behavior. Setting time and DTS data were not found to be significant (p > 0.05) between MTA with and without FGF-2. Cell proliferation and differentiation increased significantly (p < 0.05) with FGF-2 mixed MTA. After siRNA transfection with FGFR, the proliferation and differentiation behavior of the hDPCs appreciably decreased when cultured on an MTA/FGF-2 composite. In contrast, no significant amounts (p > 0.05) of ALP and OC were secreted by hDPCs seeded on MTA. The current study shows that CS substrate plays a key role in odontoblastic differentiation of hDPCs through FGFR and modulate of ERK/MAPK activation and MTA/FGF-2 enhanced the higher expression of hDPCs proliferation and osteogenic differentiation as compared to pure MTA cement.

並列關鍵字

Human dental pulp cell ； calcium silicate cement ； Mineral trioxide aggregate ； fibroblast growth factor receptor ； MAPK ； osteogenesis

參考文獻

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矽酸鈣材料結合纖維母細胞生長因子促進硬組織修復協同效應探討

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