Skeletal Muscle Engineering for the Craniofacial Region

项目摘要/摘要 面部毁容会对一个人的生活质量产生毁灭性的影响。大约26万美国人 每年都有重建手术，例如使用组织瓣来矫正先天性和后天的 头面部缺陷。大体积颅面肌组织缺损的处理仍然是一个挑战。 多达四分之一的患者因皮瓣失败、组织排斥和可获得性有限而重复手术。 仍有一部分患者的治疗完全失败，导致持续 面部毁容、经济负担和社会融合带来的挑战。产生的能力 包含患者自身细胞的头面部肌肉组织将提供更可预测的、改变生活的 对严重问题的治疗。以前的研究已经证明了头面部肌肉的重要性-- 衍生细胞和支架用于颅面骨骼肌工程。 该项目计划通过种植3D打印的仿生支架来设计颅面部肌肉组织 猪颅面肌源性细胞。目标是(1)产生有关使用 颅面肌源性细胞群用于肌肉组织的形成；(2)产生三维仿生 支持头面部肌肉发育的支架；(3)生物工程头面部肌肉组织 植入。我们的愿景是，颅面软组织缺损的永久修复可以通过以下方式实现 植入精密工程的自体颅面骨骼肌组织。假设是这样的 成功的头面部肌肉组织工程应用将结合自体颅面骨骼 肌源性细胞、3-D打印仿生支架、机械负荷的应用和胰岛素样生长 胰岛素样生长因子-1(IGF-1)，一种对肌肉细胞增殖和分化至关重要的生长因子。方法是 利用肌肉干细胞(卫星细胞)丰富猪颅面肌源性细胞群 为了再生。这些细胞将被种植到3D打印的仿生支架中，并受到载荷和IGF- 1促进再生，促进肌纤维肥大，形成适合植入的组织 进入头面部区域。目标1是生产3D打印的仿生支架，并评估 支架内丰富的猪颅面肌源性细胞群。目标2是确定 工程化猪颅面部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内交付。目标3是确定工程化的再生能力 猪颅面部骨骼肌组织的体外和体内实验。该项目的最终目标是生产一种 三者联合应用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)三维打印仿生支架；(3) 应用外部因素(机械负荷和IGF-1)。拟议研究的结果将 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.