Macrolide Accumulation by Host Cells in the Gingiva

牙龈中宿主细胞的大环内酯积累

• 批准号: 7660635
• 负责人: JOHN D WALTERS
• 金额: $ 22.5万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7660635
• 关键词: Actinobacillus actinomycetemcomitans; Active Biological Transport; Address; Adult; Adverse effects; Antibiotics; Azithromycin; Bacteria; Biological Assay; Cell Line; Cells; Clarithromycin; Clinical; Clinical Research; Connective Tissue; Debridement; Drug or chemical Tissue Distribution; Effectiveness; Epithelial Cells; Etiology; Exhibits; Fibroblasts; Foundations; Future; Gingiva; Gingival Crevicular Fluid; Goals; HL-60 Cells; Human; Immune response; In Vitro; Incidence; Individual; Infection; Inflammation; Inflammatory; Invaded; Kinetics; Liquid substance; Macrolide Antibiotics; Macrolides; Periodontal Attachment Loss; Periodontal Diseases; Periodontal Pocket; Periodontitis; Phagocytes; Pharmaceutical Preparations; Plant Roots; Play; Porphyromonas gingivalis; Process; Property; Radiolabeled; Recurrence; Regulation; Reporting; Research; Role; Serum; Signal Transduction; Substrate Specificity; Surface; System; Temperature; Therapeutic; Tooth Loss; Treatment outcome; Work; antimicrobial; base; chemotherapy; cost; design; drug distribution; insight; killings; neutrophil; novel therapeutic intervention; oral pathogen; pathogen; public health relevance; radiotracer; scaling and root planing; soft tissue; uptake

DESCRIPTION (provided by applicant): Aggressive and recurrent forms of periodontitis are associated with infections by A. actinomycetemomitans and P. gingivalis. The ability of these pathogens to invade epithelial cells and phagocytes makes them difficult to eliminate by conventional scaling and root planing alone. However, the use of systemic antibiotics in conjunction with scaling and root planing significantly enhances clinical and microbiological treatment outcomes. While not widely prescribed by periodontists, azithromycin and clarithromycin are potentially useful because they are taken up by host cells and have favorable antimicrobial activity. Both agents can potentially attain higher levels in gingiva than in serum. Preliminary studies suggest that active transport plays an important role in this process. Cultured gingival fibroblasts, epithelial cells and PMN-like HL-60 cells appear to take up clarithromycin by a concentrative and saturable transport mechanism. Clarithromycin uptake by fibroblasts is temperature-dependent and exhibits Michaelis-Menten kinetics, suggesting the involvement of an energy-dependent transport system. We hypothesize that gingival fibroblasts, epithelial cells and PMNs possess active transporters that accumulate macrolides. These transporters play a beneficial role by enhancing macrolide levels in the soft tissue wall of the periodontal pocket, in phagocytes and in gingival crevicular fluid. Since fibroblasts are a relatively large cellular compartment of the gingiva, macrolide transporters could permit these cells to function as drug reservoirs that enhance and sustain macrolide levels in gingival connective tissue and in gingival crevicular fluid. In epithelial cells and PMNs, these transporters could facilitate elimination of invasive pathogens by enhancing intracellular macrolide accumulation. The objective of this proposal is to study mechanisms by which macrolides are transported and concentrated by host cells in the gingiva. Specific aim 1 is to characterize transport systems by which gingival fibroblasts, epithelial cells and PMNs take up these agents, using radiolabeled macrolides to assay transport activity. [Specific aim 2 is to assess the pharmacological benefits of intracellular macrolide accumulation by host cells (including effects on drug distribution). Specific Aim 3 is to assess the killing of intracellular A. actinomycetemcomitans and P. gingivalis by macrolides in cultured PMNs and gingival epithelial cells]. The broad goal of this work is to provide a rational basis for selecting the most effective agents for periodontal antimicrobial chemotherapy. Ultimately, this work could contribute to reductions in tooth loss and the cost burden of treating periodontitis. PUBLIC HEALTH RELEVANCE: This proposal will study mechanisms by which macrolide antibiotics are transported and concentrated by cells in human gingiva. Exploiting cellular accumulation of macrolides could enhance the elimination of invasive periodontal pathogens that are difficult to treat with conventional scaling and root planing. By providing a rational basis for selecting an antibiotic for periodontal antimicrobial chemotherapy, this work could ultimately contribute to reductions in tooth loss and the cost burden of treating destructive periodontitis.

描述（由申请人提供）：牙周炎的侵略性和复发形式与曲霉曲霉和牙龈疟原虫的感染有关。这些病原体侵入上皮细胞和吞噬细胞的能力使它们难以通过传统的缩放和根策划来消除。然而，使用全身性抗生素与缩放和根策划结合使用显着增强了临床和微生物治疗结果。阿奇霉素和克拉霉素虽然没有被牙周医生广泛开处方，但可能有用，因为它们被宿主细胞吸收并具有有利的抗菌活性。两种药物在牙龈中的水平都比血清中的水平更高。初步研究表明，主动运输在此过程中起着重要作用。培养的牙龈成纤维细胞，上皮细胞和PMN样HL-60细胞似乎通过一种浓缩且可饱和的转运机制吸收了克拉霉素。成纤维细胞的克拉霉素摄取是温度依赖性的，并且表现出Michaelis-Menten动力学，这表明依赖能量的运输系统参与。我们假设牙龈成纤维细胞，上皮细胞和PMN具有积累大环内酯群的活性转运蛋白。这些转运蛋白通过在牙周口袋的软组织壁，吞噬细胞和牙龈刺耳的液体中增强大花环的水平来发挥有益的作用。由于成纤维细胞是牙龈的一个相对较大的细胞室，因此大环内酯类转运蛋白可以使这些细胞充当药物储层，可在牙龈结缔组织和牙龈结缔组织和牙龈填充丝裂液中增强和维持大花环水平。在上皮细胞和PMN中，这些转运蛋白可以通过增强细胞内大花环积累来促进侵袭性病原体。该提案的目的是研究通过牙龈中的宿主细胞运输和浓缩大花环的机制。具体目的1是表征牙龈成纤维细胞，上皮细胞和PMN的运输系统，使用放射性标记的大花环来分析运输活性。 [具体目的2是评估宿主细胞细胞内大花环积累的药理益处（包括对药物分布的影响）。具体目的3是评估大环内酯类培养的PMN和牙龈上皮细胞的细胞内拟南芥和牙龈疟原虫的杀死]。这项工作的广泛目标是为选择牙周抗菌化疗的最有效药物提供合理的基础。最终，这项工作可能导致减少牙齿脱落和治疗牙周炎的成本负担。 公共卫生相关性：该提案将研究大牙龈中的细胞运输和集中大环内酯类抗生素的机制。利用大环内酯类的细胞积累可以增强消除侵入性牙周病原体，这些病原体难以通过常规的缩放和根平行来治疗。通过为选择用于牙周抗菌化学疗法的抗生素提供合理的基础，这项工作最终可能有助于减少牙齿脱落和治疗破坏性牙周炎的成本负担。