Kinetics of neo-vascularization during pulp regeneration

我们已经证明，从头牙髓再生可以通过干细胞为基础的方法实现。而 这些进展标志着患者牙髓再生的最终临床实践，但存在一个关键问题 阻碍该领域的进展：缺乏及时的血管/血管生成，导致不一致的结果， 缩小适用于这种做法的情况（仅限于根尖开放的未成熟牙齿）。另夕h 牙髓再生过程中新血管形成的动力学是未知的。没有这些知识，很难 进入下一级纸浆再生。 在这里，我们建议使用血管生成诱导的牙髓干细胞（DPSC）和非血管生成诱导的牙髓干细胞（DPSC）的组合。 诱导DPSC，以促进牙髓再生过程中的新血管形成。最重要的是，我们设计了一个 新的研究模型，使我们能够进行实时动态检查的细胞过程中， 在这种方法下的血管化。 总体假设：血管生成诱导的DPSC和非诱导的DPSC的组合允许 加速和稳定的新血管形成，从而与宿主血管系统的及时血液灌注可以 并且可以达到更完全的纸浆再生。我们将利用我们新设计的 牙齿碎片皮褶窗室模型，管模型，以及我们完善的牙齿碎片 模型来检验假设。下面是具体目标。 目标1。目的：探讨新设计的血管化技术在牙髓再生中的动力学作用。 研究模型。 ·使用成为内皮样细胞的血管生成诱导的DPSC（DPSC-EC）的组合 和未诱导的DPSC用于新血管形成。 ·使用牙碎片皮褶窗室模型来研究增强 血管化和血液灌注从宿主脉管系统到工程化脉管系统的真实的时间。 目标二。目的：探讨血管生成机制和长期血管稳定性， 再生纸浆 ·研究牙髓中工程化血管/血管生成和吻合的分子机制 采用牙体缺损皮褶窗室模型进行再生。 ·检查由DPSC-EC加DPSC形成的工程化脉管系统的存活和长期稳定性 以及牙齿碎片模型中再生牙髓的质量/数量。 该项目的成功将使该领域更接近临床应用，并可能 建立了广泛应用于临床牙髓病学的技术。

We have demonstrated that de novo pulp regeneration can be achieved via stem cell-based approaches. While such progress signifies the ultimate clinical practice of pulp regeneration on patients, one critical issue exists hindering the progress in this field: lack of timely vasculo/angiogenesis rendering inconsistent outcomes and narrowing the cases suitable for such practice (limited to immature teeth with wide open apex). Additionally, the kinetics of neo-vascularization during pulp regeneration is unknown. Without such knowledge, it is difficult to advance to the next level of pulp regeneration. Here, we propose to use combination of angiogenically induced dental pulp stem cells (DPSCs) and non- induced DPSCs to enhance the neo-vascularization during pulp regeneration. Most importantly, we designed a new study model that allows us to conduct real-time kinetic inspection of the cellular process during neo- vascularization under this approach. Overall hypothesis: Combination of angiogenically induced DPSCs and non-induced DPSCs allows accelerated and stabilized neo-vascularization thereby timely blood perfusion with the host vascular system can occur and more complete pulp regeneration can be reached. We will take the advantage of our newly designed tooth fragment skin-fold window chamber model, tube model, as well as our well-established tooth fragment model to test the hypothesis. Below are the Specific Aims. Aim 1. To Investigate the kinetics of the neo-vascularization of pulp regeneration using newly designed study models. • To use combination of angiogenically induced DPSCs that become endothelial-like cells (DPSC-ECs) and non-induced DPSCs for neo-vascularization. • To use tooth fragment skin-fold window chamber model to investigate the enhancement of vascularization and blood perfusion from the host vasculature to the engineered vasculature in real time. Aim 2. To investigate the neo-vascularization mechanism and long-term neo-vascular stability of regenerated pulp. • To examine molecular mechanisms of engineered vasculo/angiogenesis and anastomosis during pulp regeneration using tooth fragment skin-fold window chamber model. • To examine survival and long-term stability of engineered vasculature formed by DPSC-ECs plus DPSCs as well as the quality/quantity of the regenerated pulp in the tooth fragment model. The success of this project will allow this field to move closer to clinical applications, and potentially establishing a technology widely used in clinical endodontics.