Developmentofan New Implant System Attached with Tissue Engineered Periodontal-Ligament

本研究建立了Hertwig上皮根鞘来源的人牙周膜上皮细胞(Malassez上皮细胞：Me)的无血清培养体系，并用无血清生长实验研究了成纤维细胞生长因子对ME和人牙周膜成纤维细胞(PLF)的促有丝分裂作用。此外，我们通过逆转录-聚合酶链式反应(RT-PCR)、聚合酶链式反应(RT-PCR)、核糖核酸酶保护试验(RPA)和免疫组织化学方法证实了成纤维细胞生长因子受体(FGFR)在ME中的表达和在PLF中的表达。FGFR3-(IIIb)、FGFR3-(IIIc)和FGFR4。免疫组织化学。对成纤维细胞生长因子-7/KGF的分析表明，成纤维细胞生长因子-7/KGF免疫反应阳性产物主要分布在细胞浆中。PLF细胞。结果表明，FGFR2-(IIIb)系统在Me的生长和分化中起重要作用。此外，内源性产生的FGFs对ME和PLF的生长和分化具有旁分泌调节作用。

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.