Cell survival in engineered skeletal muscle: The role of complement

摘要 由于创伤、肿瘤切除或先天性畸形造成的肌肉损失对患者和他们的 一家人。目前的治疗方案包括创造性地重新排列皮肤和肌肉瓣以缓解 畸形。然而，当活动的皮瓣组织收缩的百分比时，这些方法通常是次优的， 萎缩和/或死亡，功能与原始组织不同。长期以来，该领域一直在设想一种治疗方式 使用患者自己的细胞创建组织移植，然后可以移植到患者体内进行修复 形式和功能。然而，不幸的是，这种新疗法仍有许多障碍需要克服。 计划在临床上广泛采用。一个紧迫且被低估的障碍是低存活率 我们推测，植入的细胞是被对植入的移植物的先天免疫反应杀死的。一位少校 天然免疫的效应机制是补体(CP)系统。早期CP炎症的程度 事件杀死TECs内的大量细胞仍然是组织中一个重要的悬而未决的问题- 工程学领域。即使可以避免由于早期炎症导致的细胞死亡，也没有足够的血管 支持。产生具有快速吻合潜力的预血管化TECs的技术将 显著提高细胞存活率和植入率。这两个突出的事件是密切相关的。 在没有早期血管支持或早期血管支持的情况下减少早期炎症 大量的炎症反应仍然会导致细胞死亡和修复失败。因此，我们的中心假设 也就是说，CP效应通路的调节将通过减少炎症直接促进TEC的存活 细胞损伤，并通过促进新生血管。在这项提案中，我们阐明了一系列调查， 导致了一种使技术能够通过调节早期的细胞化植入物的植入 炎症过程和促进新生微血管床。我们的实验室发明了新的策略和 针对早期炎症反应的治疗学。此外，我们最近开发了一部小说 无支架技术构建自组织血管内皮细胞-成纤维细胞 (规格)。这些经过改造的结构一旦被植入，就可以很快地注入血液。vbl.使用 炎症、血管生成和吻合口的已知机制作为我们的指南，我们将系统地测试 这些创新技术使用特定的抑制剂和在肌肉下植入基因工程小鼠 模特。预期结果将提供一个可行的解决方案，从而极大地推动该领域向前发展 组织工程面临的主要障碍--移植细胞的快速血管形成和存活 还有纸巾。

Abstract Muscle loss due to trauma, tumor resection or congenital malformation is devastating to the patient and their family. Current treatment regimens involve creative rearrangement of skin and muscle flaps to mitigate the deformity. However, these approaches are often sub-optimal as a percentage of mobilized flap tissues contract, atrophy and/or die and do not function like the original tissues. The field has long envisioned a treatment modality where using the patient's own cells to create tissue grafts, that can then be transplanted into a patient to restore form and function. However, unfortunately, there are numerous hurdles yet to be overcome for this new treatment plan to be broadly adopted in the clinic. One urgent and underappreciated hurdle is the poor survival rate of implanted cells which we hypothesize are killed by the innate immune response to the implanted graft. A major effector mechanism of innate immunity is the complement (Cp) system. The extent that early Cp inflammatory events kill a substantial number of cells within the TECs remains an important unanswered question in the tissue- engineering field. Even if cell death due to early inflammation can be avoided, there is insufficient vascular support. Techniques to generate pre-vascularized TECs which have rapid anastomotic potential would substantially improve cell survival and engraftment. These two highlighted events are intimately linked. Reduction of early inflammation without early vascular support or early vascular support in the presence of a substantial inflammatory response will still result in cell death and failed repair. Therefore, our central hypothesis is that; modulation of Cp effector pathways will promote survival of TEC by reducing inflammation, direct cell injury, and by promoting neovascularity. In this proposal we articulate a line of investigation that will lead to an enabling technology to improve the engraftment of cellularized implants by modulating the early inflammatory process and promoting nascent microvascular beds. Our lab has invented novel strategies and therapeutics that target the early inflammatory response. Additionally, we have recently developed a novel scaffold-free technology to generate Self-organizing Pre-vascularized Endothelial-fibroblast Constructs (SPECs). These engineered constructs can become perfused with blood very quickly once implanted. Using known mechanisms of inflammation, angiogenesis and anastomosis as our guide, we will systematically test these innovative technologies using specific inhibitors and genetically engineered mice in a sub-muscular implant model. The expected outcomes would significantly move the field forward by providing a workable solution to the principal hurdles facing tissue engineering - rapid vascularization and survival of implanted cells and tissues.