Developmentofan New Implant System Attached with Tissue Engineered Periodontal-Ligament

本研究建立了人牙周韧带源性上皮细胞（Epithelial rests of Malassez：ME）的无血清培养体系，采用无血清生长实验研究了成纤维细胞生长因子（FGF）对ME和人牙周韧带源性成纤维细胞（PLF）的促有丝分裂作用。此外，我们还通过逆转录-聚合酶链反应（RT-PCR）、PCR-Southern杂交、核糖核酸酶保护试验（RPA）和免疫组化检测证实了FGF受体（FGFR）在ME中的表达和FGF在PLF中的表达，FGF-1和FGF-7/角质形成细胞生长因子（KGF）均刺激ME细胞生长。FGF-1和FGF-2均刺激PLF的细胞生长。RT-PCR和PCR-Southem分析显示ME表达FGFR 2-（IIIb）mRNA，但不表达FGFR 1-（IIIc）、FGFR 2-（IIIc）、FGFR 3-（IIIb）、FGFR 3-（IIIc）和FGFR 4的mRNA。免疫组织化学。FGF-7/KGF的分析显示免疫反应性FGF-7/KGF主要在细胞质中检测到。PLF细胞。PCR-Southem和RPA结果显示，PLF表达FGF-7/KGF mRNA，提示FGF-FGFR 2-（IIIb）系统在ME的生长和分化中起重要作用，内源性FGF在ME和PLF的生长和分化中起旁分泌相互调节作用。

In this study, we have developed a serum-free culture system of human periodontal ligament-derived epithelial cells (Epithelial rests of Malassez : ME) derived from Hertwig's epithelial root sheath.The mitogenic effect of fibroblast growth factor (FGF) in the ME and human periodontal ligament-derived fibroblast cells (PLF) were investigated by serum-free growth assay. In addition, we demonstrated the expression of FGF receptor (FGFR) in the ME and the expression of FGF in the PLF by reverse transcription-polymerase chain reaction (RT-PCR), PCR-Southem hybridization, ribonuclease protection assay (RPA) and immunohistochemical examinaijon.Both FGF-1 and FGF-7/keratinocyte growth factor (KGF) stimulated cell growth of the ME.On the other hand, both FGF-1 and FGF-2 stimulated cell growth of the PLF.RT-PCR and PCR-Southem analysis revealed that the ME expressed FGFR2-(IIIb) mRNA but not mRNAs for FGFR1 -(IIIc), FGFR2-(IIIc), FGFR3-(IIIb), FGFR3-(IIIc) and FGFR4. An immunohistochemical. analysis of FGF-7/KGF revealed that immunoreactive FGF-7/KGF were detected predominantly in the cytoplasm of the. PLF cells. PCR-Southem and RPA also revealed that the PLF expressed FGF-7/KGF mR.NA.These results suggest that the FGF-FGFR2-(IIIb) system plays an important role in the growth and differentiation of ME.In addition, endogenously produced FGFs work as paracrine interactive regulators of the growth and differentiation between ME and PLF.