Van der Pauw M T, Van den Bos T, Everts V, Beertsen W
Department of Periodontology, Academic Center for Dentistry Amsterdam, University of Amsterdam, The Netherlands.
J Periodontol. 2000 Jan;71(1):31-43. doi: 10.1902/jop.2000.71.1.31.
Although it is claimed that enamel matrix-derived proteins (EMP) can be used to promote new attachment formation around periodontally involved teeth, the underlying biological mechanism is not understood. It was the aim of the present study to investigate the effects of EMP on the behavior of human periodontal ligament (HPLF) and gingival fibroblasts (HGF) in vitro, with special focus on their attachment properties, the expression of alkaline phosphatase (ALP) activity, the release of transforming growth factor (TGF)beta1, and their proliferative rate.
Fibroblast populations were obtained from 10 individuals with a healthy periodontium and cultured in chemically defined medium on culture plates coated with EMP, purified collagen type I, or their respective vehicles. Experiments were performed in the absence of serum for periods up to 48 hours.
It was shown that HGF barely attached and spread on EMP-coated substrata, whereas HPLF attached and spread within 24 hours. However, when cultured on purified collagen type I, both cell types showed rapid attachment and spreading. Furthermore, the expression of ALP activity was significantly enhanced under the influence of EMP, especially in HPLF. HPLF and HGF both released significantly higher levels of TGFbeta1 in the presence of EMP. EMP did not influence 3H-thymidine incorporation by HPLF and HGF.
Our results indicate that HPLF and HGF respond differently to EMP. A more rapid attachment of HPLF to this substratum might contribute, during the initial stages of periodontal healing, to selective outgrowth and colonization of exposed root surfaces in vivo.
尽管有人声称牙釉质基质衍生蛋白(EMP)可用于促进牙周病累及牙齿周围新附着的形成,但其潜在的生物学机制尚不清楚。本研究的目的是在体外研究EMP对人牙周膜成纤维细胞(HPLF)和牙龈成纤维细胞(HGF)行为的影响,特别关注它们的附着特性、碱性磷酸酶(ALP)活性的表达、转化生长因子(TGF)β1的释放及其增殖率。
从10名牙周健康个体获取成纤维细胞群体,在涂有EMP、纯化的I型胶原或其各自载体的培养板上,于化学成分明确的培养基中培养。在无血清条件下进行长达48小时的实验。
结果显示,HGF在涂有EMP的基质上几乎不附着和铺展,而HPLF在24小时内即可附着和铺展。然而,当在纯化的I型胶原上培养时,两种细胞类型均显示出快速的附着和铺展。此外,在EMP的影响下,ALP活性的表达显著增强,尤其是在HPLF中。在有EMP存在的情况下,HPLF和HGF均释放出显著更高水平的TGFβ1。EMP不影响HPLF和HGF对3H-胸腺嘧啶核苷的掺入。
我们的结果表明,HPLF和HGF对EMP的反应不同。在牙周愈合的初始阶段,HPLF对该基质更快的附着可能有助于体内暴露根面的选择性生长和定植。