Skeletal Muscle Engineering for the Craniofacial Region

项目总结/摘要 面部毁容会对一个人的生活质量产生毁灭性的影响。约26万美国人 每年有重建程序，如使用组织瓣，以纠正先天性和后天性 颅面缺损大体积颅面肌组织缺损的管理仍然是一个挑战。 多达四分之一的患者由于皮瓣失败，组织排斥和有限的可用性而重复手术 组织;并且仍然有一部分患者治疗完全失败，导致继续 面部毁容、经济负担和融入社会的挑战。能够产生 含有患者自身细胞的颅面肌肉组织将提供更可预测、改变生活的方法 治疗严重的问题。以前的研究已经证明了颅面肌的重要性- 衍生的细胞和支架用于工程化颅面骨骼肌。 该项目提出通过在3D打印的仿生支架上种植 猪颅面肌源性细胞。目标是（1）产生关于使用 颅面肌源性细胞群用于肌肉组织的形成;（2）产生三维仿生 支持颅面肌肉发育的支架;（3）生物工程颅面肌肉组织， 置入我们的愿景是，颅面软组织缺损的永久性修复可以通过以下方式实现： 植入精密工程自体颅面骨骼肌组织。前提是 成功的颅面肌肉组织工程应用将包括自体颅面骨骼肌， 肌源性细胞，3D打印仿生支架，机械载荷的应用和胰岛素样生长 因子1（IGF-1），一种对肌肉细胞增殖和分化重要的生长因子。采取方法是 用肌肉干细胞（卫星细胞）富集猪颅面肌来源的细胞群， 为了再生。这些细胞将被接种到3D打印的仿生支架中，并接受负荷和IGF-1的刺激。 1以改善再生并促进肌纤维肥大，从而产生适合于植入的组织 进入颅面区域目的1是生产3-D打印仿生支架并评估 在支架内富集猪颅面肌来源的细胞群。目标2：确定 工程化猪颅面骨骼肌组织对机械刺激和IGF-1的反应 在定制的生物反应器中输送。目的3是确定工程化的再生能力， 猪颅面骨骼肌组织的体外和体内研究。该项目的最终目的是生产一个 功能性颅面组织修复颅面软组织缺损 创新元素：（1）颅面肌源性细胞群;（2）3D打印仿生支架;（3） 施加外部因素（机械负荷和IGF-1）。拟议研究的成果将 广泛影响整个人体的骨骼肌工程应用。

PROJECT SUMMARY/ABSTRACT Facial disfigurement can have devastating effects on one’s quality of life. Approximately 0.26 million Americans per annum have reconstructive procedures, such as the use of tissue flaps, to correct congenital and acquired craniofacial defects. Management of large volumetric craniofacial muscle tissue defects remains a challenge. Up to one quarter of patients have repeat procedures due to flap failure, tissue rejection and limited availability of tissue; and there remains a subset of patients for whom treatment completely fails resulting in continued facial disfigurement, financial burden and challenges with societal integration. The ability to generate craniofacial muscle tissue containing the patient’s own cells would provide a more predictable, life-changing treatment for a serious problem. Previous work has demonstrated the importance of craniofacial muscle- derived cells and scaffolds for engineering the craniofacial skeletal muscles. The project proposes to engineer craniofacial muscle tissue by seeding 3-D printed biomimetic scaffolds with porcine craniofacial muscle-derived cells. The goals are to (1) generate new knowledge on the use of craniofacial muscle-derived cell populations for the formation of muscle tissue; (2) produce 3-D biomimetic scaffolds to support craniofacial muscle development; (3) bioengineer craniofacial muscle tissue for implantation. The vision is that permanent restoration of craniofacial soft tissue defects can be achieved by implantation of precision-engineered autologous craniofacial skeletal muscle tissue. The hypothesis is that successful craniofacial muscle tissue engineering applications will incorporate autologous craniofacial skeletal muscle-derived cells, 3-D printed biomimetic scaffolds, application of mechanical load and insulin-like growth factor 1 (IGF-1), a growth factor important for muscle cell proliferation and differentiation. The approach is to enrich porcine craniofacial muscle-derived cell populations with muscle stem cells (satellite cells) responsible for regeneration. The cells will be seeded into 3-D printed biomimetic scaffolds and subjected to load and IGF- 1 to improve regeneration and promote muscle fiber hypertrophy leading to a tissue suitable for implantation into the craniofacial region. Aim 1 is to produce 3-D printed biomimetic scaffolds and assess response of an enriched porcine craniofacial muscle-derived cell population within the scaffolds. Aim 2 is to determine response of engineered porcine craniofacial skeletal muscle tissue to mechanical stimulation and IGF-1 delivered within customized bioreactors. Aim 3 is to determine the regenerative capability of engineered porcine craniofacial skeletal muscle tissue in vitro and in vivo. This project ultimately aims to produce a functional craniofacial tissue for restoration of craniofacial soft tissue defects through the combination of three innovative elements: (1) craniofacial muscle-derived cell populations; (2) 3-D printed biomimetic scaffolds; (3) application of external factors (mechanical load and IGF-1). The outcomes of the proposed research will broadly impact skeletal muscle engineering applications for the entire human body.