The phenotypic modulation of fibroblasts and its application for periodontal therapy

本研究建立了成纤维细胞表达碱性磷酸酶(ALP)的培养体系，以了解成纤维细胞的表型调控及其在牙周组织工程治疗中的应用。抗坏血酸(ASA)可诱导单层培养的人牙龈成纤维细胞碱性磷酸酶(ALP)活性和蛋白表达。高剂量转化生长因子-β(TGFR-β)促进细胞生长，抑制AsA对碱性磷酸酶活性的上调，低剂量促进碱性磷酸酶的诱导，但不刺激细胞增殖。将单层细胞暴露于添加AsA、高转化生长因子-β水平和10%血清的培养液中，可形成多层细胞。将这些多层膜转移到含有AsA和低水平转化生长因子-β的培养液中，可显著上调碱性磷酸酶的活性。这些结果表明，AsA和转化生长因子-β联合应用可促进成纤维细胞多层形成和碱性磷酸酶的诱导。我们的结果提示成纤维细胞表型调节可能应用于牙周组织工程治疗。

In this study, we developed a culture system in which fibroblasts express alkaline phosphatase (ALP), in order to understand the phenotypic modulation of fibroblasts and its application for the periodontal tissue-engineering therapy. Ascorbic acid (AsA) induced ALP activity and protein in human gingival fibroblasts in conventional monolayer culture. Transforming growth factor-β (TGF-β) at high doses caused cell growth and inhibited the up-regulation by AsA of ALP activity, whereas the low doses promoted ALP induction without stimulating proliferation. Exposure of cells in monolayer to the medium supplemented with AsA, high TGF-β levels, and 10% serum resulted in the formation of multilayers. Transfer of these multilayers to the medium containing AsA and low TGF-β levels caused a marked up-regulation of ALP activity. These results demonstrate that a combination of AsA and TGF-β promoted multilayer formation and ALP induction in fibroblasts. Our results suggest a possible application of fibroblast phenotypic modulation for the periodontal tissue-engineering therapy.