2012-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/647498摘要：牙齒的發育與其它器官一樣都是在胚胎時期經由上皮細胞與間葉細胞相互作用之後而形成，因此探討牙齒再生最大的目標是找尋相關的機制以作為探討器官再生的模式。藉由探討牙胚上皮細胞與間葉細胞作用之機轉將可作為探討器官再生的調控機制。本研究計劃的目的乃是探討牙胚上皮細胞與間葉細胞之相互作用導引分化之調控機制，以建立器官再生研究模式。本研究計劃為三年計劃，特定目標如下:目標一：探討不同生長因子骨形成蛋白(BMP2, BMP4s)及纖維母細胞生長因子(FGF9,FGF10)對於牙胚上皮細胞與間葉細胞相互作用之調控機制。目標二：探討不同生長因子骨形成蛋白(BMP2, BMP4s)及纖維母細胞生長因子(FGF9,FGF10)作用於牙胚上皮細胞與間葉細胞後，在老鼠體內牙齒再生與發育的影響。目標三：探討不同生長因子骨形成蛋白(BMP2, BMP4s)及纖維母細胞生長因子(FGF9,FGF10)作用於牙胚上皮細胞與間葉細胞後，在迷你猪體內牙齒再生與發育的影響。研究方法 如下第一年(1) 進行體外研究, 分離小鼠牙胚上皮細胞與間葉細胞並重新組合後，實驗組有四組分別加入不同的骨形成蛋白(BMP2, BMP4s)及纖維母細胞生長因子(FGF9, FGF10)，第一組加BMP2, 第二組加BMP4, 第三組加FGF9, 第四組加FGF10, 另有一組控制組,進行體外立體培養實驗，對於牙胚上皮細胞與間葉細胞互動與分化的影響。(2)利用RT-PCR 探討上皮細胞與間葉細胞經由不同生長因子骨形成蛋白(BMP2, BMP4s)及纖維母細胞生長因子(FGF9, FGF10) 作用對於牙胚上皮細胞與間葉細胞分化表現的影響。第二年利用老鼠動物模式進行體外實驗;將分離之牙胚上皮細胞與牙胚間葉細胞重新組合後，實驗組有四組分別加入不同的骨形成蛋白(BMP2, BMP4s)及纖維母細胞生長因子(FGF9,FGF10)，第一組加BMP2, 第二組加BMP4, 第三組加FGF9, 第四組加FGF10, 另有一組控制組,進行體外立體培養實驗，植入大鼠體內比較不同生長因子骨形成蛋白(BMP2,BMP4s)及纖維母細胞生長因子(FGF9, FGF10)對於牙齒生長速度與形態發育的影響。第三年利用迷你猪動物模式進行體外實驗; 分離胚胎迷你猪的牙胚上皮細胞與間葉細胞，將分離之牙胚上皮細胞與牙胚間葉細胞重新組合，實驗組有四組分別加入不同的骨形成蛋白(BMP2, BMP4s)及纖維母細胞生長因子(FGF9, FGF10)，第一組加BMP2, 第二組加BMP4, 第三組加FGF9, 第四組加FGF10, 另有一組控制組,進行體外立體培養實驗，並植入迷你猪體內比較不同生長因子骨形成蛋白(BMP2, BMP4s)及纖維母細胞生長因子(FGF9, FGF10)對於牙齒生長速度與形態發育的影響。<br> Abstract: Tooth development is originated from the cross-talk between embryonic epithelial cellsand mesenchymal cells which is similar to other organs. Therefore, the most meaningful goalsof tooth regeneration is to find the important mechanism of interactions between epithelialcells and mesenchymal cells for building up the study model for other organ regeneration.The purposes of this study are to investigate the interactions between embryonic dentalepithelial cells and mesenchymal cells for the development of tooth and build up a model forfurther study of organ regeneration.This is a three year project, the specific aims are as follows:Aim 1. To investigate the effects of different factors including bone morphogenicproteins (BMP2, BMP4) and fibroblast growth factors (FGF9, FGF10) )separately on cell-cell interactions between dental epithelial cells andmesenchymal cellsAim 2. To investigate the effects of different factors including bone morphogenicproteins (BMP2, BMP4) and fibroblast growth factors (FGF9, FGF10) )separately on epithelial-mesenchymal interactions and tooth regeneration in rats.Aim 3. To investigate the effects of different factors different factors including bonemorphogenic proteins (BMP2, BMP4) and fibroblast growth factors (FGF9,FGF10) ) separately on epithelial-mesenchymal interactions and toothregeneration in mini pigs.The materials and methods are as follows:In the 1st year(1) In vitro study by isolating embryonic dental epithelial cells and mesenchymal cellsand then recombinate these two kinds of cells for 3D cell culture and added withdifferent factors including bone morphogenic proteins (BMP2, BMP4) and fibroblastgrowth factors (FGF9, FGF10) ) separately to investigate the effects of differentfactors on cell-cell interactions between dental epithelial cells and mesenchymal cells(2) Using RT-PCR to investigate the effects of different factors including bonemorphogenic proteins (BMP2, BMP4) and fibroblast growth factors (FGF9, FGF10)separately on the differentiation of dental epithelial cells and mesenchymal cellsIn the 2nd yearIn vivo study by using rats as animal model. By isolating the embryonic dental epithelialcells and mesenchymal cells from rats and then recombinate these two kinds of cells for 3Dcell culture and added with different factors including bone morphogenic proteins (BMP2,BMP4) and fibroblast growth factors (FGF9, FGF10) ) separately and then transplanted intorats to see the effects of different factors on the growth rate of tooth regeneration in rats.. .In the 3rd yearIn vivo study by using mini pigs as animal model. By isolating the embryonic dentalepithelial cells and mesenchymal cells from embryo mini pigs and then recombinate these twokinds of cells for 3D cell culture and added with different factors including bone morphogenicproteins (BMP2, BMP4) and fibroblast growth factors (FGF9, FGF10) separately and thentransplanted into mini pigs to see the effects of different factors on the growth rate of toothregeneration in mini pigs.. .上皮細胞與間葉細胞互動牙齒再生骨形成蛋白纖維母細胞生長因子epithelial-mesenchymal cell interactionstooth regenerationbone morphogenic proteinsfibroblast growth factors