Effects of growth/differentiation factor-5 on human periodontal ligament cells
Effects of growth/differentiation factor-5 on human periodontal ligament cells
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DOI：
10.1034/j.1600-0765.2003.00695.x
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发表时间：
2003-12-01
影响因子：
3.5
通讯作者：
Izumi, Y
中科院分区：
文献类型：
--
作者：
Nakamura, T;Yamamoto, M;Izumi, Y
目的：生长/分化因子-5（GDF-5）是转化生长因子-β超家族的成员，与骨形态发生蛋白具有密切的结构关系，在骨骼、肌腱和韧带的形态发生中起关键作用。编码GDF-5的mRNA在牙胚发育过程中也有表达，特别是在牙囊组织中。虽然这表明GDF-5参与牙槽骨和牙周膜、牙骨质和牙根的形成，但GDF-5在成年期这些组织中的生理作用仍不清楚。因此，我们研究GDF-5对培养的人牙周膜细胞（HPDL）的影响。材料和方法：HPDL细胞从健康的个人牙周膜获得。四唑还原试验用于细胞增殖试验。通过测量405 nm处的吸光度估计碱性磷酸酶（ALP）活性。逆转录-聚合酶链反应（RT-PCR）和北方分析在培养的HPDL细胞的基因表达。结果：RT-PCR检测到GDF-5及其受体在HPDL细胞中的表达。rhGDF-5在10-1000 ng/ml浓度范围内可显著降低HPDL细胞ALP活性（p < 0.05）。尽管北方分析显示rhGDF-5刺激的HPDL细胞中胶原α 2（I）的基因表达几乎没有变化，但rhGDF-5剂量依赖性地增强细胞增殖。这种增殖效应持续16 d。结论：rhGDF-5可能通过影响牙周组织细胞外基质的代谢，为牙周组织的修复和再生提供环境。
Objectives: Growth/differentiation factor-5 (GDF-5), a member of the transforming growth factor-beta superfamily, shows a close structural relationship to bone morphogenetic proteins and plays crucial roles in skeletal, tendon, and ligament morphogenesis. The mRNA encoding GDF-5 is also expressed during odonto-genesis, especially in dental follicle tissue. While this suggests that GDF-5 participates in the formation of alveolar bone and the periodontal ligament, cementum, and dental root, the physiologic role of GDF-5 in these tissues in adulthood remains unclear. We therefore investigated GDF-5 effects upon cultures of human periodontal ligament (HPDL) cells.Material and methods: HPDL cells were obtained from healthy periodontal ligaments of individuals. Tetrazolium reduction assay was carried out for cell proliferation assay. Alkaline phosphatase (ALP) activity was estimated by measuring light absorbance at 405 nm. Reverse transcription-polymerase chain reaction (RT-PCR) and northern analysis were performed for gene expression in cultured HPDL cells. Sulfated glycosaminoglycan (sGAG) synthesis was evaluated by histochemical staining and a quantitative dye-binding method.Results: Expression of GDF-5 and its receptor was demonstrated in HPDL cells by RT-PCR. ALP activity in HPDL cells was significantly decreased by addition of rhGDF-5 at 10-1000 ng/ml (p < 0.05). Although northern analysis showed little change in gene expression for collagen alpha2(I) in rhGDF-5-stimulated HPDL cells, rhGDF-5 dose-dependently enhanced cell proliferation. This proliferative effect persisted for 16 d. Alcian blue staining and dye-binding assays indicated that sGAG synthesis was enhanced by rhGDF-5.Conclusion: rhGDF-5 may provide an environment fostering periodontal healing or regeneration by affecting extracellular matrix metabolism.