Mechanisms of Pro-Resolving Mediators in Periodontal Regeneration

项目摘要 不受控制的炎症是组织工程、再生和重建的主要障碍。 患病和受伤的组织导致进一步的组织损伤、组织结疤和纤维化。干细胞 活性被促进炎症的持续信号破坏，而特异性抗炎信号 增强干细胞活性。在成功再生中，间充质干细胞承担抗炎作用， 表型专门的促分解脂质介质（SPMS），包括脂氧素A4（LXA 4）， 干细胞炎症反应炎症的消退是一个活跃的生化和代谢过程， 而不仅仅是由SPM介导的炎症的被动终止。SPM激活伤口愈合， 组织再生而不是纤维化和瘢痕形成并直接改善骨愈合和再生， 包括牙周炎。人牙周膜干细胞释放SPMs，包括脂氧素，以调节 免疫调节和促愈合特性。表征人体SPM的仿生特性 由于缺乏合适的大型动物模型，迫切需要一种经过验证的大型动物 再生模型，以测试SPM翻译到人类的治疗潜力。我们的目标是确定 控制局部炎症和增强间充质干细胞的再生途径 分化成结缔组织，包括骨。中心假设是， 炎症途径和介质可以促进牙周器官（骨、牙骨质）的再生 和牙周膜）通过指导干细胞表型、增殖和分化。在本申请中， 我们将使用一个大型动物模型来解剖SPM途径导致牙周膜干细胞控制 重生的机会在本研究中，我们将：1：为约克郡小型猪生产SPM提供直接证据 猪牙周膜干细胞（mpPDLSC），通过测定mpPDLSC的脂质介质谱; 2： 通过测定mpPDLSC增殖和对SPM的反应来测定小型猪中的干细胞功能， 以及合成酶表达、信号传导途径和SPM受体表达的测定，以及3： 用SPM局部实验证明SPM促进小型猪干细胞介导的牙周再生 单独或与移植的、离体扩增的、 小型猪干细胞这些研究的结果将促进我们对剂量和剂量的实际临床知识， 在组织再生中递送脂氧素，鉴定潜在的新分子靶点，并进一步开发和 表征大型动物模型，以测试用于翻译成人类口腔的基于干细胞的新策略， 颅面组织再生该研究小组由牙周再生专家组成， 生物化学和大型动物模型。

Project Summary Uncontrolled inflammation is a major impediment to tissue engineering, regeneration and reconstruction of both diseased and injured tissues resulting in further tissue injury, tissue scarring and fibrosis. Stem cell activity is disrupted by persistent signals promoting inflammation, whereas specific anti-inflammatory signals enhance stem cell activity. In successful regeneration, mesenchymal stem cells assume an anti-inflammatory phenotype. Specialized Proresolving lipid Mediators (SPMS), including Lipoxin A4 (LXA4), attenuate the dental stem cell inflammatory response. Resolution of inflammation is an active biochemical and metabolic process, not merely a passive termination of inflammation, mediated by SPMs. SPMs activate wound healing with tissue regeneration instead of fibrosis and scarring and directly improve bone healing and regeneration, including in periodontitis. Human periodontal ligament stem cells release SPMs, including lipoxin, to regulate immunomodulatory and pro-healing properties. Characterizing the biomimetic properties of SPMs in humans is hampered by a lack of suitable large animal models. There is a critical need for a validated large animal regeneration model to test therapeutic potential of SPMs for translation to humans. Our goal is to determine the pathways to regeneration that control local inflammation and enhance mesenchymal stem cell differentiation into connective tissues, including bone. The Central Hypothesis is that resolution of inflammation pathways and mediators can promote regeneration of the periodontal organ (bone, cementum and periodontal ligament) by directing stem cell phenotype, proliferation and differentiation. In this application, we will use a large animal model to dissect the SPM pathways leading to periodontal ligament stem cell control of regeneration. In this proposal, we will: 1: Provide direct evidence for SPM production by Yorkshire miniature pig periodontal ligament stem cells (mpPDLSC) by determining the lipid mediator profile of mpPDLSC; 2: Determine stem cell function in miniature pig by determining mpPDLSC proliferation and response to SPMs, and determination of synthetic enzyme expression, signaling pathways and SPM receptor expression, and 3: Demonstrate SPM enhanced stem cell mediated periodontal regeneration in Miniature Pigs using SPM local delivery to enhance periodontal regeneration alone or in combination with transplanted, ex vivo expanded miniature pig stem cells. Results from these studies will advance our practical clinical knowledge of dose and delivery of lipoxins in tissue regeneration, identify potential new molecular targets, and further develop and characterize a large animal model to test novel stem cell-based strategies for translation to human oral and craniofacial tissue regeneration. The research team comprises experts in periodontal regeneration, biochemistry, and large animal models.