Mechanisms of Pro-Resolving Mediators in Periodontal Regeneration

项目摘要 炎症失控是组织工程、再生和重建的主要障碍。 病变和损伤的组织都会导致进一步的组织损伤、组织疤痕形成和纤维化。干细胞 活动被持续的促进炎症的信号干扰，而特定的抗炎信号 增强干细胞活性。在成功的再生过程中，间充质干细胞具有抗炎作用。 表型。包括脂氧素A4(LXA4)在内的专门的促分解脂质介体(SPMS)可以削弱牙齿 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， 不只是被动地终止炎症，由SPM介导。SPM通过以下方式促进伤口愈合 组织再生而不是纤维化和疤痕形成，并直接促进骨愈合和再生， 包括牙周炎。人牙周膜干细胞释放SPM，包括脂氧素，以调节 具有免疫调节和促进愈合的特性。SPM在人体内的仿生特性表征 由于缺乏合适的大型动物模型而受到阻碍。迫切需要一种经过验证的大型动物 再生模型，以测试SPM移植到人类的治疗潜力。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。中心假设是解决 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)，通过指导干细胞的表型、增殖和分化。在此应用程序中， 我们将使用一个大型动物模型来解剖导致牙周膜干细胞控制的SPM通路。 重生的力量。在这项提案中，我们将：1：为约克郡微型公司生产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： 通过检测mpPDLSC的增殖和对SPMS的反应来确定小型猪的干细胞功能， 以及合成酶表达、信号通路和SPM受体表达的测定，以及3： SPM局部增强干细胞介导的小型猪牙周再生 单独或与移植、体外扩增联合应用促进牙周再生 微型猪干细胞。这些研究的结果将促进我们对剂量和 脂氧素在组织再生中的传递，识别潜在的新分子靶点，并进一步开发和 描述一种大型动物模型，以测试基于干细胞的新策略，将其转化为人类口腔和 颅面组织再生。研究团队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.