低强度脉冲超声协同DFCs/BMSCs诱导牙周组织再生的实验研究

Seed cells start the process of tissue regeneration, then recruit massive autologous cells to participate in this regeneration. However, as the common seed cell of periodontal tissue engineering, Dental Follicle Cells(DFCs) show poor resources; Hence, having further numerous autologous cells recruited in the periodontal tissue regeneration is the key element. Our previous studies have showed that the co-culture of DFCs and Bone Marrow Mesenchymal Stem Cells(BMSCs) might make for tissue regeneration, and Low-Intensity Pulsed Ultrasound(LIPUS) provides synergistic effect on this process. However,the exact mechanism involved is still undefined. Based on our previous Natural Science Foundation of China(NSFC) research, we construct the co-culture system of DFCs/BMSCs and related animal models to analyze the proliferation and differentiation of DFCs and BMSCs cell-interaction in the co-culture , and to observe cell homing of autologous cells ,with the technology of tissue engineering and molecular biology. Furthermore, we’ll investigate the exact molecular mechanism of biological events referring to LIPUS enhancing the activity of seed cells, boosting cell-interaction, inducing cells homing, suppressing the LPS-meditated periodontal local inflammation and promoting the periodontal tissue regeneration. Our team combine this project with the tissue engineering technology and the medical philosophy of noninvasive ;We intend to determine the role of the cell-interaction played in the co-culture of DFCs and BMSCs. Morever, we’ll investigate the combined application of DFCs/BMSCs and the synergistic effect of LIPUS during periodontal tissue regeneration. The research shall devote a brand new thought for the periodontal tissue regeneration.

种子细胞启动组织再生过程并募集更多自体细胞参与。DFCs是牙周组织工程常用种子细胞，来源有限，如何解决并诱导更多自体细胞参与牙周组织再生极为关键。项目前期发现DFCs与BMSCs共培养可能更利于牙周组织再生，LIPUS也具有协同作用，但LIPUS如何协同DFCs与BMSCs诱导牙周组织再生的作用不清楚。本研究拟在前期国家自然科学基金研究基础上，采用组织工程、分子生物学等实验方法，构建DFCs/BMSCs共培养体系和相应细胞、动物模型，研究DFCs/BMSCs共培养下细胞相互作用对增殖、诱导分化的影响，观察对自体细胞的募集效应，探讨LIPUS增强细胞功能活性、细胞间相互作用，促进细胞募集，抑制LPS介导的牙周组织局部炎症，促进牙周组织再生等生物学机制。项目紧密结合组织工程技术和无创理念，以期阐明DFCs/BMSCs联合应用和LIPUS的协同效应在牙周组织再生的作用，为牙周组织再生开辟新思路。

种子细胞启动组织再生过程并募集更多自体细胞参与。DFCs是牙周组织工程常用种子细胞.，来源有限，如何解决并诱导更多自体细胞参与牙周组织再生极为关键。项目前期发现DFCs与BMSCs共培养可能更利于牙周组织再生，LIPUS也具有协同作用，但LIPUS如何协同DFCs与BMSCs诱导牙周组织再生的作用不清楚。本项目采用transwell实验、siRNA技术、免疫荧光技术、real time PCR、western blot、扫描电子显微镜、荧光素酶报告系统等实验研究发现，LIPUS通过TWIST1促进了SDF1的表达和分泌，从而促进了牙周膜细胞等的迁移和归巢。此外，LIPUS对LPS诱导的牙周膜细胞及巨噬细胞炎症因子IL-6、IL-8的表达具有抑制效果，后续研究发现与NFκB信号通路有关。LIPUS对牙周膜细胞的氧化应激和内质网应激反应均有抑制作用，Nrf2及IRE1α信号通路参与其中。LIPUS可以通过促进细胞外基质的合成及细胞成骨分化促进牙周组织的再生。本项目的研究意义在于进一步阐明了LIPUS对细胞分化、免疫调节、细胞归巢和组织再生的分子机制，为LIPUS辅助的牙周组织工程临床应用提供了科学依据。