An immunomodulatory approach to improve oral cavity wound healing

项目概述：腭裂患者(1：1000活产)都接受了腭裂修复；然而，60%的患者 包括口鼻瘘(ONF)形成在内的包膜并发症。ONF的形成会导致摄食不良和 演讲。腭裂修复术涉及将腭部粘膜瓣旋转到裂隙中，愈合受到物理因素的影响。 卡尔的舌头创伤和细菌滋生的环境。目前，腭裂修复手术缺乏 再生性重建选择，由于ONF需要多次重新修复，增加了成本和发病率。 外科医生使用同种异体真皮植入物，尽管有艾滋病毒/普里恩病毒传播的风险，但仅作为一种屏障 减少ONF。我们的团队已经开发出一种新的ONF的小鼠表型，并鉴定了一种独特的免疫- 在促进再生的计划中指导ONF愈合过程的调节性方法。FTY720，一款活跃的 生物脂，靶向鞘氨醇途径，优先吸引促再生单核细胞和巨噬细胞- ES以促进ONF的愈合。我们的长期目标是开发一种免疫再生方法来改善PAL- 吃了伤口愈合。本应用的总体目标是确定ONF愈合和 使用FTY720纳米纤维以促进再生的方式调整单核细胞和巨噬细胞的迁移。这个 中心假设是炎性单核细胞和巨噬细胞是口腔中的主要反应。 空洞伤口愈合和FTY720的释放优先吸引促再生单核细胞和Mac- 能促进伤口愈合的罗弗斯。FTY720信号通过鞘氨醇途径，增加 促进再生的单核细胞和巨噬细胞的募集。建议进行这项研究的理由是 对ONF形成过程中的炎症反应进行全面和机械性的了解将 提供旨在改变促再生单核细胞迁移的治疗靶点。在强劲的前期指导下- 没有数据，包括一篇证明在FTY720纳米纤维交付后ONF显著减少的论文， 这一假设将通过追求两个具体目标来检验：1)确定需求、时机和机制- 在ONF形成过程中炎症反应的自然贡献；2)设计一种纳米纤维支架来 使用免疫再生AP提供FTY720的受控释放以促进口腔伤口的愈合 试探一下。在目标1中，我们将确定关键的炎性细胞募集对修复ONF，确定重新- ONF后单核细胞停滞及细胞因子和修复基因通路的变化 队形。在目标2中，我们将测试FTY720的最佳传递策略，FTY720纳米纤维定向的能力 应用和不使用单核细胞促进创面再生修复，并测定FTY720纳米粒子对创面愈合的影响 细胞因子和修复基因通路上的纤维。所提出的研究在机械方面具有创新性。 描述了允许测试免疫调节策略的ONF小鼠模型中的单核细胞迁移。 这项拟议的研究意义重大，因为它表明FDA批准的药物FTY720被批准用于治疗多发性硬化症 硬化症，可以快速进入临床试验，以减少ONF的形成。

PROJECT SUMMARY: Cleft palate patients (1:1000 live births) all undergo palate repair; however, 60% de- velop complications including oronasal fistula (ONF) formation. ONF formation leads to poor feeding and speech. Cleft palate repair involves rotating palate mucosal flaps into the cleft and healing is affected by physi- cal trauma from the tongue and a bacteria laden environment. Currently, cleft palate repair surgeries lack a regenerative reconstructive option, require multiple re-repairs due to ONF and increase cost and morbidity. Surgeons use allograft dermal implants, despite the risk of HIV/prion transmission, to act solely as a barrier to reduce ONF. Our group has developed a novel murine phenocopy of ONF, and identified a unique immuno- modulatory approach to directing the ONF healing process in a pro-regenerative program. FTY720, an active biolipid, targets the sphingosine pathway to preferentially attract pro-regenerative monocytes and macrophag- es to improve ONF healing. Our long-term goal is to develop an immunoregenerative approach to improve pal- ate wound healing. The overall objective in this application is to determine the mechanism of ONF healing and use FTY720 nanofibers to tune the monocyte and macrophage migration in a pro-regenerative fashion. The central hypothesis is that inflammatory monocytes and macrophages are the predominant response during oral cavity wound healing and that delivery of FTY720 preferentially attracts pro-regenerative monocytes and mac- rophages leading to improved wound healing. FTY720 signals through the sphingosine pathway, increasing the recruitment of pro-regenerative monocytes and macrophages. The rationale for the proposed research is that a comprehensive and mechanistic understanding of the inflammatory response during ONF formation will provide therapeutic targets aimed at altering pro-regenerative monocyte migration. Guided by strong prelimi- nary data, including a paper demonstrating significant reduction in ONF following FTY720 nanofiber delivery, the hypothesis will be tested by pursuing two specific aims: 1) Determine the requirement, timing and mecha- nistic contributions of the inflammatory response during ONF formation; 2) Engineer a nanofiber scaffold to provide controlled delivery of FTY720 to improve oral cavity wound healing using an immunoregenerative ap- proach. In Aim 1 we will identify the critical inflammatory cell recruitment to healing ONF, determine the re- quirement of monocytes, and identify alteration in the cytokines and reparative gene pathways following ONF formation. In Aim 2, we will test the optimal FTY720 delivery strategy, the ability of FTY720 nanofiber to direct pro-regenerative ONF wound healing with and without monocytes, and determine the effects of FTY720 nano- fiber on the cytokines and reparative gene pathways. The proposed research is innovative by mechanistically describing monocyte migration in an ONF murine model that allows testing of immunomodulatory strategies. The proposed research is significant because it that FTY720, an FDA-approved drug approved to treat multiple sclerosis, could be fast tracked to clinical trials to reduce ONF formation.