Regeneration of periodontal structures through the recruitment of regulatory lymp

描述(由申请人提供)：越来越清楚的是，牙周炎不仅以病原性感染为特征，而且更是以免疫稳态的丧失为特征。在不完全阻断局部和全身感染的免疫反应的情况下再生牙周组织的策略将是理想的。基于对调节性T细胞生物学的最新见解，以及它们在牙周炎中的缺失，我们假设调节性T细胞招募到牙周组织将解决疾病症状并促进再生环境。为此，我们开发了一种初步的控释制剂，用于传递调节性T细胞招募因子(CCL-22)。在我们的牙周病小鼠模型中，该制剂的牙周内口袋给药增加了调节性T细胞向牙周组织的迁移，并导致了疾病症状的改善。为了进一步了解调节性T细胞(Treg)治疗牙周炎的机制，并开发更具临床可行性的治疗方法，我们的目标如下。具体目标一：研究Treg Recruiting制剂的治疗机制。在这里，我们将在小鼠牙周炎的实验模型中，基于我们目前的治疗方案，研究调节性T细胞迁移到牙周组织并滞留在牙周组织中的动力学。此外，我们将跟踪Treg相关因子、促炎和抗炎介质以及导致硬组织和软组织破坏的因素的表达，以观察Treg的存在如何影响它们在牙周组织中的表达。最后，为了测试Treg-Recruiting配方的再生性能，我们将使用活体动物成像技术测量治疗前后的牙槽骨水平。具体目标二：CCL-22微粒的合理设计，可自主产生复杂的释放特性。我们的数据强烈表明，多次注射后，Treg Recruiting(CCL-22)微粒的初步配方具有治疗效果。为了进行临床上更理想的治疗，我们建议使用新的数学模型来设计两种不同的配方，根据两种假设的复杂趋化因子释放时间表产生精确定义的释放行为。特定目的III：检测血管活性肠肽作为天然细胞分泌CCL-22的潜在介体。我们的初步数据显示，血管活性肠肽(VIP)治疗上调内源性Treg募集趋化因子CCL-22在小鼠牙龈组织中的表达。由于VIP从产品开发的角度来看很有吸引力，我们将系统地研究VIP的治疗潜力，并通过长期、受控的给药方式将其输送到牙周袋。 公共卫生相关性：这项研究与公共健康相关，因为它打算解决当今最紧迫的口腔健康问题-牙周炎，估计影响到7800万美国人。这种疾病不仅会导致牙齿脱落，还会增加心血管疾病、糖尿病、呼吸系统疾病和早产的发生率。此外，我们的方法可能与其他疾病相关，如颞下颌关节骨关节炎。

DESCRIPTION (provided by applicant): It is becoming clear that periodontitis is not only characterized by pathogenic infection, but even more so by a loss of immunological homeostasis. Strategies toward regenerating the periodontium without completely blocking immune responses against local and systemic infections would be ideal. Based on recent insights into regulatory T cell biology, and their absence in periodontitis, we hypothesize that the recruitment of regulatory T cells to the periodontium will resolve disease symptoms and promote a regenerative milieu. To this end, we have developed a preliminary controlled release formulation for the delivery of a regulatory T cell recruiting factor (CCL-22). In our periodontal disease mouse model, intra-periodontal pocket delivery of this formulation increased regulatory T cell migration to the periodontium and led to the amelioration of disease symptoms. To further understand the mechanisms for regulatory T cell (Treg) therapy in periodontitis and develop more clinically viable therapeutics, we purpose the following aims. Specific Aim I: Investigation into the therapeutic mechanisms of Treg recruiting formulations. Herein we will examine the kinetics of regulatory T cell migration to, and residence in, the periodontium based on our current therapeutic formulation, in an experimental model of murine periodontitis. Furthermore, we will track the expression of Treg-associated factors, pro- and anti-inflammatory mediators, as well as factors that lead to hard and soft tissue destruction, to observe how Treg presence influences their presentation in the periodontium. Finally, to test the regenerative properties of Treg-recruiting formulations, we will use a live animal imaging technique to measure alveolar bone levels before, and after, treatment. Specific Aim II: Rational design of CCL-22 microparticles engineered to autonomously produce complex release characteristics. Our data strongly suggests a therapeutic effect associated with our preliminary formulation of Treg recruiting (CCL-22) microparticles after multiple injections. To make a more clinically- desirable therapy, we propose to use new mathematical models to engineer two different formulations that produce precisely defined release behavior according to two hypothetical schedules of complex chemokine delivery. Specific Aim III: Examination of VIP as a potential mediator of CCL-22 secretion by native cells. Our preliminary data shows treatment with vasoactive intestinal peptide (VIP) up-regulates the expression of endogenous Treg recruiting chemokine CCL-22 in gingival tissues of mice. Because VIP is attractive from a product development standpoint, we will investigate the therapeutic potential of VIP both systemically and through long-lasting, controlled delivery to the periodontal pocket. PUBLIC HEALTH RELEVANCE: The research has relevance to public health as it intends to address the most pressing oral health concern today, periodontitis, affecting an estimated 78 million Americans. This disease not only leads to tooth loss, but increased incidence of cardiovascular diseases, diabetes, respiratory diseases, and premature births. Further, our approach may have relevance to other disorders as osteoarthritis of the temporo-mandibular joint.