ORAL MUCOSAL CELL RESPONSES TO DENTAL RESIN COMPONENTS

口腔粘膜细胞对牙科树脂成分的反应

• 批准号: 2387051
• 负责人: GEORGE S SCHUSTER
• 金额: $ 21.09万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2387051
• 关键词: biomaterial compatibility; biomaterials; calcium flux; composite resins; connective tissue cells; dental prosthesis; enzyme activity; human tissue; lipid metabolism; lysophospholipids; mucosal immunity; oral mucosa; phosphatidate; phosphatidylcholines; phosphatidylethanolamines; phospholipase A2; phospholipase C; phospholipase D; polymethacrylate; tissue /cell culture; unsaturated fatty acids

DESCRIPTION (Adapted from the investigator's Abstract): Substances that elute from dental resins and composite materials may alter the growth and metabolism of cells with which they come in contact. The substances themselves may affect the cells or they may be hydrolyzed by tissue enzymes and the products of hydrolysis affect the cells. The cell changes may relate to growth, membrane composition and membrane-related functions, many of which are related to cell sterol and phospholipid composition. Two widely used components of resins, dimethylaminoethyl methacrylate (DMAEMA) and hydroxyethyl methacrylate (HEMA), produce such changes and will serve as prototype compounds for the study. The former affects cell neutral lipid and phospholipid metabolism while the latter appears to only alter neutral lipid metabolism. In the case of neutral lipid metabolism, the major effects of both compounds are decreases in cholesterol and increase in cholesterol esters, with some changes in other neutral lipid classes. In the case of phospholipids, there are changes in phosphatidycholine (PC) and phosphatidylethanolamine (PE) with accumulation of a methylated precursor of PC, dimethylphosphatidylethanolamine (DMPE). Changes in sterols and levels of methylated phospholipid alter a variety of membrane-associated functions. The specific aims of the current proposal are to examine 1) hydrolysis of the methacrylates and their effects on cell lipid metabolism of oral epithelial and connective tissue cells, as well as a non-keratinizing epithelial cell type and a non-oral fibroblast cell type, 2) cellular phospholipase activity, and 3) cell Ca2+ fluxes. Cultures of the various cells will be exposed to the methacrylates and the patterns of lipid metabolism analyzed by thin-layer chromatography. Alterations in the phospholipid pathways in response to DMAEMA will be examined biochemically, since the different cell types respond in a different manner quantitatively. The ability of cells and tissue esterases to hydrolyze the materials will also be assessed using cell homogenates and commercial tissue esterases. Since methylated phospholipids and cell sterol composition can affect calcium fluxes, changes in [CA2+]i response to the materials will be determined using spectrofluorimetery. Phospholipase A2, C and D( PLA2, PLC, PLD) activities are affected by methylated phospholipids and by cell calcium, and products of their activity can provide not only precursors for the methylated phospholipid, but also components of the intracellular cell signaling pathways. These activities will be measured using radiolabeled substrates. These studies will provide information concerning the mechanism by which cells metabolize methacrylates and how the compounds can affect cells.

描述（改编自研究者摘要）： 从牙科树脂和复合材料中提取的杂质可能会改变生长， 与它们接触的细胞的代谢。 的物质 它们本身可以影响细胞，或者它们可以被组织酶水解， 并且水解产物影响细胞。 细胞变化可能 与生长、膜组成和膜相关功能有关， 与细胞固醇和磷脂组成有关。 两 广泛使用的树脂成分，甲基丙烯酸二甲氨基乙酯（DMAEMA） 和甲基丙烯酸羟乙酯（HEMA），产生这种变化，将作为 用于研究的原型化合物。 前者影响细胞中性脂质 和磷脂代谢，而后者似乎只改变中性 脂质代谢 在中性脂质代谢的情况下， 这两种化合物的作用是降低胆固醇和增加 胆固醇酯，其他中性脂质类也有一些变化。 在 在磷脂的情况下，磷脂酰胆碱（PC）和 磷脂酰乙醇胺（PE）与积累的甲基化前体 PC，二甲基磷脂酰乙醇胺（DMPE）。 固醇和水平的变化 甲基化磷脂改变了多种膜相关功能。 本提案的具体目标是审查1）水解 甲基丙烯酸酯及其对口腔黏膜细胞脂代谢的影响 上皮和结缔组织细胞，以及非角质化 上皮细胞类型和非口腔成纤维细胞类型，2）细胞 磷脂酶活性; 3）细胞Ca ~（2+）流。 各种文化 细胞将暴露于甲基丙烯酸酯和脂质的模式 通过薄层色谱法分析代谢。 的改变 将对响应DMAEMA的磷脂途径进行生化检查， 因为不同的细胞类型以不同的方式定量响应。 细胞和组织酯酶水解材料的能力将 也可以使用细胞匀浆和商业组织酯酶进行评估。 由于甲基化磷脂和细胞固醇的组成可以影响 钙通量，[CA 2 +]i响应材料的变化将是 用荧光分光光度法测定。 磷脂酶A2、C和D（PLA 2，PLC， PLD）活性受甲基化磷脂和细胞因子的影响。 钙及其活性产物不仅可以提供 甲基化磷脂，还有细胞内细胞的成分， 信号通路 这些活动将使用放射性标记的 印刷受体. 这些研究将提供有关该机制的信息， 细胞通过什么代谢甲基丙烯酸酯，以及化合物如何影响 细胞