Personalized bioprinting technology for de novo PDL regeneration

摘要 恒牙撕脱伤是最严重的牙科损伤之一。牙周膜 (PDL)在撕脱过程中被破坏和污染，以及再植后的结果 撕脱的牙齿是不好的。使用PDL干细胞(PDLSCs)的细胞治疗为我们提供了 再生牙撕脱后牙周韧带的机会。最近对大型动物的研究表明 承诺以细胞为基础的治疗撕脱牙。然而，目前还缺乏 系统设计和部署尖端组织工程技术类型， 最终将导致为新的PDL开发个性化的医疗方法 撕脱牙齿的再生。在这里，我们建议采用新开发的两步法 技术，即使用生物合成球体和干细胞的个性化3D生物打印- 介导的PDL再生。第一步涉及生产含有PDLSCs的生物墨水 包裹在微球中，称为PDLSC-生物合成球。第二步使用一个 定制的3D生物打印机设计了特定的算法，可以打印一层PDLSC- 生物复合体沿着其自然表面附着在去污染和剥离的牙根上 地形，在整个根上具有期望的和均匀的PDL再生厚度 浮出水面。我们假设，撕脱牙齿上丢失的PDL可以用 使用PDLSC生物合成球进行个性化生物打印。在目标1中，我们将优化生成 PDLSC-生物合成微球及其印制后的生物学行为 在体外与已建立的SCID小鼠模型进行牙根表面移植。在目标2中，我们将利用 一种研究功能性PDL体内再生的简易小型猪模型。我们 预计PDLSC生物合成球和个性化3D生物打印的结合 技术将在根部产生一致的PDL再生，这将产生初步的 这些数据为未来R01应用中更广泛的大型动物研究奠定了基础。这个 重大的临床影响是有可能在 田纳西州再生医学研究所(Tennessee Institute of Regenerative Medicine，TennIRM)用于再植脱位牙齿 取得成功的长期有利结果。未来的努力将包括建立干细胞 银行，以便利和保护这种医疗疗法的细胞来源。

ABSTRACT Avulsion of permanent teeth is one of the most serious dental injuries. The periodontal ligament (PDL) is damaged and contaminated during avulsion and the outcome after replantation of the avulsed teeth is poor. Cell-based therapy using PDL stem cells (PDLSCs) provides us with the opportunity to regenerate the PDL of avulsed teeth. Recent large animal studies have shown promise of cell-based therapies for managing avulsed teeth. However, there is a lack of systematic design and deployment of the type of cutting-edge tissue engineering technology that would ultimately lead to development of a personalized medicine approach for the de novo PDL regeneration of avulsed teeth. Here we propose to incorporate a newly developed, two-step technology, namely the use of Biosynspheres and personalized 3D-bioprinting for stem cell- mediated PDL regeneration. The first step involves the production of bio-ink containing PDLSCs encapsulated in microspheres, termed PDLSC-Biosynspheres. The second step employs a custom-made 3D-bioprinter designed with specific algorithms that can print a layer of PDLSC- Biosynspheres onto the decontaminated and denuded tooth root following its natural surface topography, with a desired and uniformed thickness for PDL regeneration on the entire root surface. We hypothesize that the lost PDL on avulsed teeth can be regenerated de novo using personalized bioprinting with PDLSC-Biosynspheres. In Aim 1, we will optimize the generation of PDLSC-Biosynspheres and characterize their biological behaviors after being printed onto the root surface in vitro and with an established SCID mouse model in vivo. In Aim 2, we will utilize a simple mini-swine model to characterize the regeneration of functional PDL in vivo. We anticipate that the combination of PDLSC-Biosynspheres and personalized 3D-bioprinting technology will result in consistent PDL regeneration on the root, which will generate preliminary data that sets the stage for more extensive large animal studies in a future R01 application. The significant clinical impact is the potential to establish a Dental Trauma Management Center at the Tennessee Institute of Regenerative Medicine (TennIRM) for replanting avulsed teeth aiming at successful long-term favorable outcomes. Future endeavor will include establishing stem cell banking to facilitate and secure the cell source for such medical therapies.