Muscle progenitor cell-based implants for dynamic laryngeal muscle reconstruction

 描述(由申请人提供)：在美国，每年有数以千计的人遭受严重的失声(发音困难或失声)影响，这些人接受创伤性或肿瘤性喉部分切除术，或因声带瘫痪而遭受肌肉容量丧失。这些患者的语音恢复方案不是最理想的，因此，大多数患者都会留下永久性的失声和沟通障碍。这项应用为直接声带损伤和/或去神经损伤后恢复声带肌肉容量和功能提供了一种新的方法。结果可能会改善声带瘫痪和/或接受过半喉切除、喉皮质切除或创伤性撕脱的患者的发音恢复手术选择。这项应用的第一个目标是开发一种肌祖细胞来源的植入物(MI)，在动物模型中植入后，可以获得强大的神经支配。具体地说，将确定肌纤维运动终板表达对心肌梗死植入后神经支配的影响。为了做到这一点，我们将在体外构建并使用诱导心肌梗死肌肉表达运动终板的因子对心肌梗死进行预处理。在同种异体大鼠模型中，mis将被用来替代部分喉切除的缺损区，并将根据喉肌电图和与神经接触的运动终板的量化来确定植入后的神经状态。使用该动物植入模型，在具有和不具有喉返神经完整性的环境中，将研究MIS的结果与对照MIS的结果进行比较。这项应用的下一个目标是在定制的胶原蛋白基质中开发MI，并确定该方法是否有助于改善肌肉纤维的排列，并在用于修复喉部肌肉缺陷时增加肌肉体积。为了达到这些目标，将使用胶原聚合物创建MI，该聚合物展示胶原纤维基质的自组装和分级工程设计，以指导细胞命运，从而促进肌肉纤维的形成，其大小、肌纤维密度和排列模仿天然声带肌肉的特征。同样，使用动物模型，在有和没有喉返神经完整的环境中，胶原基MI的结果将与对照MI的结果进行比较。拟议的研究结果应该会克服目前开发用于半咽重建的功能性组织工程MCC的主要障碍-这些障碍是肌肉的神经支配不足，肌纤维的次优组织，以及肌肉与自然内收肌的不同步激发。此外，这些初步实验的结果应该会导致里程碑式的临床创新，这些创新将与语音恢复应用和全球肌肉修复概念相关。

 DESCRIPTION (provided by applicant): Devastating voice loss (dysphonia or aphonia) impacts thousands of individuals in the United States each year undergoing traumatic or oncologic partial laryngectomies, or suffering muscle volume loss due to vocal fold paralysis. Voice restoration options for these patients are suboptimal, and, as a result, most patients are left with permanent voice loss and communication impairment. This application introduces a novel approach for restoring vocal fold muscle volume and function after direct vocal fold injury and/or denervation. Results may lead to improved surgical options for voice restoration in patients who have vocal paralysis and/or have undergone hemilaryngectomies, cordectomies, or traumatic avulsions. The first goal of this application is to develop a muscle progenitor cell-derived implant (MI) that, after implantation in an animal model, receives strong innervation. Specifically, the effect of myofiber motor endplate expression on post-implantation innervation of MIs will be determined. To do this, we will fabricate and pre-treat MIs with factors in vitro that induce the MI muscle to express motor endplates. The MIs will be used to replace a partial laryngectomy defect in a syngeneic rat model, and post-implantation innervation status, based on laryngeal electromyography and quantification of motor endplates with nerve contact, will be determined. Using this animal implant model, outcomes with the study MIs will be compared to those of control MIs in environments with and without recurrent laryngeal nerve integrity. The next goal of this application is to develop a MI within a customized collagen matrix, and determine if this approach will facilitate improved alignment of muscle fibers and enhanced muscle volume when used to repair laryngeal muscle defects. To reach these goals, MIs will be created with collagen polymers exhibiting self-assembly and hierarchical engineering design of collagen fibril matrices to guide cell fate, and thereby facilitating formation of muscle fibers wih size, myofiber density, and alignment that mimics the characteristics of the native vocal fold muscle. Again, using an animal model, outcomes with the collagen based MIs will be compared to those of control MIs in environments with and without recurrent laryngeal nerve integrity. Findings from the proposed studies should overcome current major hurdles to developing a functional tissue engineered MCC for hemilaryngeal reconstruction-those hurdles being inadequate innervation of the muscle, suboptimal organization of myofibers, and asynchronous firing of the muscle with the native adductor muscle. Furthermore, results from these initial experiments should lead to landmark clinical innovations that will be relevant to both voice restoration applications, and muscle repair concepts globally.