Macrolide Accumulation by Host Cells in the Gingiva

牙龈中宿主细胞的大环内酯积累

基本信息

批准号：
7783831
负责人：
JOHN D WALTERS
金额：
$ 18.56万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-01 至 2013-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7783831
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

项目摘要

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）：侵袭性和复发性牙周炎与放线放线菌和牙龈卟啉单胞菌感染有关。这些病原体侵入上皮细胞和吞噬细胞的能力使得它们难以仅通过传统的刮治和根面平整来消除。然而，全身抗生素的使用结合洗牙和根面平整术可显着提高临床和微生物治疗效果。虽然牙周病医生没有广泛使用阿奇霉素和克拉霉素，但它们可能有用，因为它们被宿主细胞吸收并具有良好的抗菌活性。这两种药物在牙龈中的含量可能高于血清中的含量。初步研究表明主动运输在此过程中发挥着重要作用。培养的牙龈成纤维细胞、上皮细胞和 PMN 样 HL-60 细胞似乎通过集中且可饱和的转运机制吸收克拉霉素。成纤维细胞对克拉霉素的吸收具有温度依赖性，并表现出米氏动力学，表明涉及能量依赖性运输系统。我们假设牙龈成纤维细胞、上皮细胞和中性粒细胞具有积聚大环内酯类的活性转运蛋白。这些转运蛋白通过提高牙周袋软组织壁、吞噬细胞和龈沟液中大环内酯的水平而发挥有益作用。由于成纤维细胞是牙龈相对较大的细胞区室，大环内酯转运蛋白可以允许这些细胞充当药物储库，增强和维持牙龈结缔组织和龈沟液中的大环内酯水平。在上皮细胞和中性粒细胞中，这些转运蛋白可以通过增强细胞内大环内酯的积累来促进消除侵入性病原体。该提案的目的是研究牙龈中宿主细胞转运和浓缩大环内酯类药物的机制。具体目标 1 是表征牙龈成纤维细胞、上皮细胞和中性粒细胞吸收这些药物的转运系统，使用放射性标记的大环内酯来测定转运活性。 [具体目标 2 是评估宿主细胞细胞内大环内酯积累的药理学益处（包括对药物分布的影响）。具体目标 3 是评估大环内酯类药物对培养的 PMN 和牙龈上皮细胞中胞内放线放线菌和牙龈卟啉单胞菌的杀灭作用]。这项工作的总体目标是为选择最有效的牙周抗菌化疗药物提供合理的基础。最终，这项工作可能有助于减少牙齿脱落和治疗牙周炎的成本负担。公共健康相关性：该提案将研究人类牙龈细胞转运和浓缩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对传统刮牙和根面平整难以治疗的侵入性牙周病原体的消除。通过为牙周抗菌化疗选择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。