The Gene Therapy of Pulp Cell Differentiation into Odontoblasts using Growth / Differentiation Factor 11.
使用生长/分化因子 11 进行牙髓细胞分化为成牙本质细胞的基因治疗。
基本信息
- 批准号:11470406
- 负责人:
- 金额:$ 9.73万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (B).
- 财政年份:1999
- 资助国家:日本
- 起止时间:1999 至 2000
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Gene therapy is now evolving rapidly and gaining significant momentum as a therapeutic strategy to treat patients suffered from inherited disease, infectious disease, and malignant tumors in the near future. Among the nonviral techniques for gene transfer, electroporation is simple, inexpensive and safe. We investigated the applicability of in vivo electroporation of gene transfer into amputated dental pulp, using plasmid DNA expressing Growth/Differentiation factor ( Gdf) 11 as the vector. The bead soaked with recombinant GDF11 could induce the mRNA expression of Dentin sialoprotein (DSP), the differentiation marker of odontoblasts, in the dental papillae cells surrounding them, 7 days after organ culture. The conditions required for optimal in vitro (or ex vivo) electroporation were determined by green fluorescent protein (GFP) expression after the gene transfer of GDF11-GFP cDNA plasmid into dental papillae mesenchyme. The optimal Gdf11 gene transfer into the dental pulp mesenchymal cells much increased the mRNA expression of DSP locally as the recombinant GDF11 application. It demonstrated that Gdf11 cDNA plasmid transferred by electroporation might have potential for gene therapy of dental pulp cell differentiation into odontoblasts and reparative dentin formation in vivo. Therefore, for in vivo electroporation we used the same condition and the same electrode as the in vitro electroporation. There was a difficulty, however, in the approach of the electrode to the surface of the exposed pulp in the narrow cavity without any damage on the pulp tissue. The pressure and heat by the electrode resulted in the necrosis of the pulp tissue locally attached to the electrode. Further appraisal of nonviral mean such as ultrasound should be waited for more powerful and convenient gene transfer into the exposed dental pulp.
基因治疗正在迅速发展,并在不久的将来成为一种治疗遗传病、传染病和恶性肿瘤患者的治疗策略。在非病毒基因转移技术中,电穿孔是一种简单、廉价和安全的技术。本研究以表达生长/分化因子(Gdf) 11的质粒DNA为载体,探讨了在体电穿孔基因转染断牙髓的可行性。用重组GDF11浸泡的牙珠可诱导成牙细胞周围的牙乳头细胞中牙本质涎蛋白(DSP) mRNA的表达,该蛋白是成牙细胞的分化标志物。将GDF11-GFP cDNA质粒转染到牙乳头间质后,通过绿色荧光蛋白(GFP)的表达来确定体外(或离体)电穿孔的最佳条件。将Gdf11基因转染到牙髓间充质细胞后,重组Gdf11在牙髓间充质细胞中的表达量显著增加。结果表明,经电穿孔转移的Gdf11 cDNA质粒在牙髓细胞向成牙细胞分化和修复性牙本质形成的体内基因治疗中具有潜在的应用价值。因此,对于体内电穿孔,我们使用了与体外电穿孔相同的条件和相同的电极。然而,在狭窄的腔体中,电极接近暴露的牙髓表面而不损伤牙髓组织是一个困难。电极的压力和热量导致局部附着在电极上的牙髓组织坏死。进一步评估非病毒手段,如超声,应等待更强大和方便的基因转移到暴露的牙髓。
项目成果
期刊论文数量(13)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
M.Nakashima,T.Toyono,A.Akamine,A.Joyner: "Expression of growth/differentiaton factor 11, a new member of the BMP/TGFβ superfamily during mouse embryogenesis."Mech.Dev.. 80・2. 185-189 (1999)
M.Nakashima、T.Toyono、A.Akamine、A.Joyner:“小鼠胚胎发生过程中 BMP/TGFβ 超家族的新成员生长/分化因子 11 的表达。”Mech.Dev.. 80・2。 189 (1999)
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- 影响因子:0
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- 通讯作者:
M.Nakashima, T.Toyono, T.Murakami A.Akamine: "Transforming growth factor-β superfamily members expressed in rat incisor pulp."Archs oral Biol. 43-9. 745-751 (1998)
M.Nakashima、T.Toyono、T.Murakami A.Akamine:“大鼠门牙牙髓中表达的转化生长因子-β 超家族成员”。Archs 口腔生物学 43-9(1998)。
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- 影响因子:0
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- 通讯作者:
H.L.Park,C.Bai,K.A.Platt,M.P.Matise,A.Beeghly,C.c.Hui,M.Nakashima and A.L.Joyner: "Mouse Glil mutants are viable but have defects in SHH signaling in combination with a Gli2 mutation."Development. 128・8. 1593-1605 (2000)
H.L.Park、C.Bai、K.A.Platt、M.P.Matise、A.Beeghly、C.c.Hui、M.Nakashima 和 A.L.Joyner:“小鼠 Glil 突变体是可行的,但在与 Gli2 突变结合的 SHH 信号传导方面存在缺陷。”128。・8。1593-1605(2000)
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
M.Nakashima, T.Toyono, A.Akamine, A.Joyner: "Expression of growth/differentiaton factor 11, a new member of the BMP/TGF β superfamily during mouse embryogenesis."Mech.Dev.. 80-2. 185-189 (1999)
M.Nakashima、T.Toyono、A.Akamine、A.Joyner:“小鼠胚胎发生过程中 BMP/TGF β 超家族的新成员生长/分化因子 11 的表达。”Mech.Dev.. 80-2。 -189 (1999)
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
M. Nakashim,T. Toyono,A. Akamine,A. Joyner: "Expression of growth/defferntiation factor 11,a new member of the BMP/TGFβ superfamily during mouse embryogenesis."Mech. Dev.. 80・2. 185-189 (1999)
M. Nakashim,T. Toyono,Akamine,A. Joyner:“小鼠胚胎发生过程中 BMP/TGFβ 超家族的新成员的表达”。 80・2。 189 (1999)
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NAKASHIMA Misako其他文献
NAKASHIMA Misako的其他文献
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{{ truncateString('NAKASHIMA Misako', 18)}}的其他基金
Development of Optimal Cell Culture Method with Morphological Quality Assessment by DNA Damage
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- 批准号:
26293422 - 财政年份:2014
- 资助金额:
$ 9.73万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Demonstration of transdifferentiation of other tissue stem cells into dental pulp stem cells by a pulp biomarker
通过牙髓生物标志物证明其他组织干细胞转分化为牙髓干细胞
- 批准号:
20390504 - 财政年份:2008
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$ 9.73万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Ex Vivo Gene Therapy for Dentin Regeneration Using Dental Pulp Stem Cells transfected with Gdf11
使用转染 Gdf11 的牙髓干细胞进行牙本质再生的体外基因治疗
- 批准号:
15390577 - 财政年份:2003
- 资助金额:
$ 9.73万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Molecular Cloning to Dentin Morphogenetic Protein (DMP)
牙本质形态发生蛋白 (DMP) 的分子克隆
- 批准号:
07044279 - 财政年份:1995
- 资助金额:
$ 9.73万 - 项目类别:
Grant-in-Aid for international Scientific Research
Molecular Cloning of Dentin Morphogenetic Protein
牙本质形态发生蛋白的分子克隆
- 批准号:
07457460 - 财政年份:1995
- 资助金额:
$ 9.73万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
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