3D-bioprinting of sustained- and phased-release antibiotic and probiotic scaffolds to treat bacterial vaginosis

持续和分阶段释放抗生素和益生菌支架的 3D 生物打印用于治疗细菌性阴道病

• 批准号: 10580042
• 负责人: Hermann Frieboes
• 金额: $ 76.74万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-25 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10580042
• 关键词: 3-Dimensional; 3D Print; Adherence; Adverse event; Affect; Aftercare; Age; Alginates; Anaerobic Bacteria; Antibiotic Therapy; Antibiotics; Antifungal Agents; Appearance; Atopobium vaginae; Back; Bacteria; Bacterial Vaginosis; Biological Assay; Biomedical Engineering; Calibration; Cell Line; Cells; Clinical; Clinical Research; Clinical Trials; Complex; Computer Models; Data; Development; Devices; Epithelial Cells; Epithelium; Equilibrium; Experimental Models; Female; Formulation; Gardnerella; Gardnerella vaginalis; Gelatin; Goals; Growth; HIV; Health; Histopathology; Human; In Vitro; Infection; Infertility; Inflammation Mediators; Investigation; Kinetics; Lactobacillus; Link; Measurement; Measures; Medicine; Metronidazole; Microbe; Modeling; Mucous body substance; Mus; Oral; Outcome; Pelvic Inflammatory Disease; Phase; Pre-Clinical Model; Prevotella; Printing; Probiotics; Process; Proliferating; Property; Reapplication; Records; Recurrence; Regimen; Retreatment; Serum; Sexually Transmitted Diseases; Silicones; Symptoms; Syndrome; Testing; Therapeutic; Therapeutic Agents; Tissues; Topical Antibiotic; Topical application; Treatment Efficacy; Treatment Protocols; Treatment outcome; Uterus; Vagina; Woman; Work; adverse pregnancy outcome; aged; bioprinting; co-infection; cytotoxicity; delivery vehicle; design; dysbiosis; engineering design; experience; experimental study; fabrication; gut health; high risk; improved; improved outcome; in vitro Model; in vitro testing; in vivo; mathematical model; microbial; microbicide; mouse model; multidisciplinary; novel; pathogen; pre-clinical; predictive modeling; prevent; probiotic therapy; prototype; reduce symptoms; reproductive; reproductive tract; scaffold; vaginal lactobacilli; vaginal microbiome; virtual

PROJECT SUMMARY Bacterial vaginosis (BV) is a dysbiosis of the vaginal microbiome that affects ~29% of reproductive age women and is linked with higher risks of adverse pregnancy outcomes, postsurgical infections, and sexually transmitted infections. While lactobacilli typically dominate the healthy vagina, BV is characterized by low lactobacilli levels and an overgrowth of diverse anaerobic bacteria, most often including Gardnerella and Prevotella. Current oral or topical antibiotic treatments alleviate symptoms in 80% of women, at least temporarily. However, most women will experience a recurrence of BV within one-year post-treatment. A recent clinical study showed that a vaginal probiotic treatment regimen (with Lactobacillus crispatus), used following a vaginal course of antibiotics (metronidazole), significantly improved long-term treatment efficacy. Unfortunately, current topically-applied formulations require repeated administrations (once to twice daily), which can hinder female convenience and adherence to treatment, particularly when undergoing multiple weeks of treatment (required to administer both antibiotic and probiotics). In this project we will use 3D printing and computational modeling, iteratively enabled by functional investigation of prototype scaffolds in vitro and in vivo, to design long-acting products that sustain therapeutic delivery, while enabling phased-delivery of antibiotics and probiotics to the female reproductive tract. Our team brings together female reproductive tract-specific expertise in delivery vehicle design, computational modeling, and preclinical BV models. The ultimate goal is to use 3D-bioprinting to incorporate different device compartments, which sequentially release antibiotics that target anaerobic overgrowth, followed by live probiotics that restore balance back in favor of vaginal lactobacilli. In Aim 1, we will design and characterize 3D-printed silicone scaffolds that sustain antibiotic-only delivery. Aim 2 will design and evaluate 3D-printed silicone and gelatin alginate composites that sequentially release antibiotics followed by probiotics, providing a “1-2 punch” strategy to kill BV bacteria and provide a ‘healthy’ Lactobacillus alternative. Each aim will focus first on a materials-based characterization of Met-containing silicone (1A) or Met-silicone probiotic-gelatin alginate composites (2A). Measurements from in vitro release experiments will be used to develop and test computational models that predict the delivery of antibiotic (1B) or dual agents (2B) in a “virtual female reproductive tract” and ultimately in a mouse co-infection model. We will evaluate Met-silicone (1C) and Met-silicone probiotic-gelatin alginate composites (2C) for cytotoxicity to the vaginal epithelium and ability to stimulate soluble proinflammatory mediators and downstream histopathology. In our mouse model, we will measure levels of viable Gardnerella and Prevotella recovered from vaginal and uterine tissues following treatment with blank or active agent- containing 3D-printed scaffolds. If successful, this project will support the development of multipurpose platforms to prevent and treat BV as well as other female reproductive tract applications.

项目摘要 细菌性阴道病（BV）是一种阴道微生物群失调，影响约29%的育龄妇女 并且与不良妊娠结局、术后感染和性传播的风险较高有关。 感染.虽然乳酸杆菌通常占主导地位的健康阴道，BV的特点是低乳酸杆菌水平 以及各种厌氧细菌的过度生长，最常见的包括加德纳菌和普雷沃菌。当前口服 或局部抗生素治疗缓解症状，在80%的妇女，至少暂时。然而，大多数女性 将在治疗后一年内发生BV复发。最近的一项临床研究表明， 益生菌治疗方案（使用卷曲乳杆菌），在抗生素阴道疗程后使用 （甲硝唑），显著提高长期治疗疗效。不幸的是，目前的主题应用 制剂需要重复给药（每天一至两次），这可能妨碍女性的便利性， 对治疗的依从性，特别是在接受多周治疗时（需要同时给予 抗生素和益生菌）。 在这个项目中，我们将使用3D打印和计算建模，通过功能调查迭代启用 原型支架在体外和体内，设计长效产品，维持治疗输送，而 从而能够将抗生素和益生菌分阶段递送到女性生殖道。我们的团队汇集了 女性生殖道特定的专业知识，在运载工具设计，计算建模， BV模型。最终目标是使用3D生物打印来整合不同的设备隔间， 依次释放针对厌氧过度生长的抗生素，然后是恢复平衡的活益生菌 阴道乳酸杆菌的优势在目标1中，我们将设计和表征3D打印的硅胶支架， 维持只向南极运送。目标2将设计和评估3D打印硅胶和明胶藻酸盐 复合材料依次释放抗生素，然后释放益生菌，提供了杀死BV的“1-2拳”策略 细菌和提供一个'健康'乳酸菌的替代品。每个目标将首先侧重于一个基于材料的 含Met的硅氧烷（1A）或Met-硅氧烷益生菌-明胶藻酸盐复合物（2A）的表征。 体外释放实验的测量结果将用于开发和测试计算模型， 预测抗生素（1B）或双药剂（2B）在“虚拟女性生殖道”中的递送，并最终在 小鼠共感染模型。我们将评估Met-硅酮（1C）和Met-硅酮益生菌-藻酸明胶 复合物（2C）对阴道上皮细胞的细胞毒性和刺激可溶性促炎性细胞因子的能力 介质和下游组织病理学。在我们的小鼠模型中，我们将测量活加德纳菌的水平， 和在用空白或活性剂处理后从阴道和子宫组织中回收的普雷沃氏菌- 包含3D打印的支架。如果成功，该项目将支持多用途平台的开发 预防和治疗BV以及其他女性生殖道应用。