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转化生长因子-β1(TGF-β1)对人成牙本质细胞和牙髓细胞中I型和III型胶原蛋白mRNA及蛋白质合成的基线表达及体外影响

Baseline expression and effect of TGF-β1 on Type I and III collagen mRNA and protein synthesis in human odontoblasts and pulp cellsIn Vitro.

作者信息

Palosaari H, Tasanen K, Risteli J, Larmas M, Salo T, Tjäderhane L

机构信息

Institute of Dentistry, University of Oulu, Oulu, Finland.

Department of Dermatology and Collagen Research Unit Biocenter, University of Oulu, Oulu, Finland.

出版信息

Calcif Tissue Int. 2001 Feb;68(2):122-129. doi: 10.1007/BF02678151.

Abstract

Since growth factors have been suggested to regulate dentin collagen formation in response to external irritation, we investigated the effect of TGF-β1 on proα1(I) collagen mRNA expression in cultured mature human odontoblasts and pulpal fibroblasts, as well as cultured human pulp tissue, using quantitative PCR. Cultured gingival fibroblasts (GF) and osteoblasts (OB) served as controls. Also, type I collagen synthesis in cultured odontoblasts and pulp tissue, as well as type III collagen synthesis in odontoblasts, were studied by measuring respective procollagen (PINP and PIIINP) secretion into culture media with radioimmunoassay (RIA). Odontoblasts expressed significantly higher basic level of type I collagen mRNA than pulp tissue or pulp fibroblasts in culture, but markedly lower level than GF and OB cells. TGF-β1 (10 ng/ml) had negligible effects on type I collagen mRNA expression or PINP synthesis in cultured odontoblasts and pulp tissue, and PIIINP synthesis in the odontoblasts. In PF cells, the effect of TGF-β1 depended on culturing conditions; a 6-fold increase in mRNA expression was observed using serum-free medium but no effect was seen in the cells cultured with 10% FBS. In contrast, GF cells serving as controls were not markedly affected by the culture conditions, with 2-3-fold increase in mRNA expression by TGF-β1. These experiments demonstrate that mature human odontoblasts are capable of synthesizing type III collagen protein, and that TGF-β1 has negligible effect on mature human odontoblast and pulp tissue collagen expression.

摘要

由于有研究表明生长因子可响应外部刺激调节牙本质胶原蛋白的形成,我们使用定量PCR研究了转化生长因子-β1(TGF-β1)对培养的成熟人成牙本质细胞、牙髓成纤维细胞以及培养的人牙髓组织中proα1(I)胶原蛋白mRNA表达的影响。培养的牙龈成纤维细胞(GF)和成骨细胞(OB)作为对照。此外,通过放射免疫分析(RIA)测量培养基中各自前胶原(PINP和PIIINP)的分泌量,研究了培养的成牙本质细胞和牙髓组织中的I型胶原蛋白合成以及成牙本质细胞中的III型胶原蛋白合成。在培养中,成牙本质细胞表达的I型胶原蛋白mRNA基础水平显著高于牙髓组织或牙髓成纤维细胞,但明显低于GF和OB细胞。TGF-β1(10 ng/ml)对培养的成牙本质细胞和牙髓组织中的I型胶原蛋白mRNA表达或PINP合成以及成牙本质细胞中的PIIINP合成影响可忽略不计。在PF细胞中,TGF-β1的作用取决于培养条件;在无血清培养基中观察到mRNA表达增加6倍,但在含10%胎牛血清(FBS)培养的细胞中未见影响。相比之下,作为对照的GF细胞受培养条件影响不明显,TGF-β1使mRNA表达增加2 - 3倍。这些实验表明,成熟的人成牙本质细胞能够合成III型胶原蛋白,并且TGF-β1对成熟的人成牙本质细胞和牙髓组织胶原蛋白表达影响可忽略不计。

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