Winter S, Kohl A, Huppertz A, Herold-Mende C, Wiest T, Komposch G, Tomakidi P
Department of Operative Dentistry and Periodontology, Dental School, University of Heidelberg, Im Neuenheimer Feld 400, 69120, Germany.
Cell Tissue Res. 2005 Mar;319(3):467-78. doi: 10.1007/s00441-004-1026-z. Epub 2005 Jan 25.
Although the function and effects of many growth factors and extracellular matrix (ECM) molecules have been described for several periodontal tissues in vivo and in vitro, the molecular interactions involved in the communication between cells of the periodontal ligament and the alveolar bone are poorly understood. To contribute to the identification of such interactions, we have generated co-cultures (CCs) of periodontal ligament fibroblasts (PDLs) and alveolar bone cells (ABCs) and compared mRNA expression for various growth factors, ECM molecules, and matrix metalloproteinase13 (MMP13) after 1 and 2 weeks with matched mono-cultures (MCs) by reverse transcription/polymerase chain reaction. Compared with CCs of 1 week, PDLs and ABCs after 2 weeks revealed relatively high levels of all analyzed mRNAs, viz., for EGF, HGF, VEGF, TGFbeta1, collagen-I (COL1), osteonectin (ON), fibronectin (FN1), and MMP13. At week 2, when compared with MCs, CCs showed an elevation of all tested mRNAs in PDLs and ABCs, except for TGFbeta1 and FN1, which only increased in PDLs. After 1 week, when CCs were compared with MCs, mRNAs for HGF and TGFbeta1 were less abundant in PDLs and ABCs, whereas the other genes exhibited lower expression levels in only one of the cell types. Analysis of our data indicated that the expression of mRNAs for growth factors and for COL1, ON, FN1, and MMP13 was modulated in the distinct cell types with respect to culture time and culture type. The differences in the mRNA expression patterns between CCs and MCs suggest that the respective genes are involved in the molecular interactions of PDLs and ABCs.
尽管多种生长因子和细胞外基质(ECM)分子在体内和体外对几种牙周组织的功能及作用已被描述,但牙周膜细胞与牙槽骨细胞之间通讯所涉及的分子相互作用仍知之甚少。为有助于识别此类相互作用,我们构建了牙周膜成纤维细胞(PDL)与牙槽骨细胞(ABC)的共培养体系(CC),并通过逆转录/聚合酶链反应,将1周和2周后的共培养体系与相应的单培养体系(MC)进行比较,分析了多种生长因子、ECM分子及基质金属蛋白酶13(MMP13)的mRNA表达情况。与1周的共培养体系相比,2周后的PDL和ABC显示出所有分析的mRNA水平相对较高,即表皮生长因子(EGF)、肝细胞生长因子(HGF)、血管内皮生长因子(VEGF)、转化生长因子β1(TGFβ1)、I型胶原(COL1)、骨连接蛋白(ON)、纤连蛋白(FN1)和MMP13的mRNA水平。在第2周时,与单培养体系相比,共培养体系中PDL和ABC的所有检测mRNA均升高,但TGFβ1和FN1除外,这两种分子仅在PDL中增加。在第1周时,将共培养体系与单培养体系比较,PDL和ABC中HGF和TGFβ1的mRNA含量较少,而其他基因仅在其中一种细胞类型中表达水平较低。对我们数据的分析表明,生长因子以及COL1、ON、FN1和MMP13的mRNA表达在不同细胞类型中受培养时间和培养类型的调节。共培养体系与单培养体系之间mRNA表达模式的差异表明,相应基因参与了PDL和ABC的分子相互作用。