Engineering Vascularized Skeletal Muscle for Treatment of Volumetric Muscle Loss

工程血管化骨骼肌用于治疗体积性肌肉损失

• 批准号: 10158427
• 负责人: Ngan F. Huang
• 金额: --
• 依托单位: VETERANS ADMIN PALO ALTO HEALTH CARE SYS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2023-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10158427
• 关键词: Biomedical Engineering; Blood; Blood Vessels; Cell Density; Cell Line; Cell Survival; Cell Transplantation; Cicatrix; Clinical; Cues; DNA Sequence Alteration; Data; Debridement; Disease; Economic Burden; Endothelial Cells; Endothelium; Engineering; Explosion; Extracellular Matrix; Gene Delivery; Generations; Goals; Growth; Growth Factor; Gunshot wound; Health; Histologic; Impairment; In Vitro; Incidence; Injury; Lasers; Length; Measurement; Measures; Mediating; Messenger RNA; Morbidity - disease rate; Mus; Muscle; Muscle Fibers; Muscle function; Muscular Atrophy; Myoblasts; Natural regeneration; Operative Surgical Procedures; Outcome; Oxygen; Patients; Pattern; Perfusion; Physiological; Play; Proteomics; Recovery; Role; Site; Skeletal Muscle; Spectrum Analysis; Structure; Supplementation; Surgical Flaps; System; Testing; Therapeutic; Tissue Engineering; Tissues; Transfection; Transplantation; Traumatic injury; Treatment Efficacy; Vascular Endothelial Cell; Vehicle crash; Veterans; bioluminescence imaging; blood perfusion; combat; confocal imaging; cytokine; design; disability; experience; healing; improved; in vivo; injured; mechanical properties; military veteran; mouse model; muscle engineering; muscle physiology; muscle regeneration; muscle strength; muscular structure; nanofibrillar; nanopattern; nanoscale; novel strategies; repaired; scaffold; tibialis anterior muscle; transplantation therapy; volumetric muscle loss

Volumetric muscle loss (VML) is characterized by the loss of a significant portion of skeletal muscle, leading to permanent damage to muscle structure and function. VML results from major traumatic injury, and it is becoming increasingly more frequent in military Veterans as a result of roadside explosions, gunshot wounds, and motor vehicle crashes. VML contributes to long-term disability and $400 billion in economic burden in the US annually. Traumatic injuries leading to VML are associated with impaired endogenous muscle regeneration and revascularization capacity. Current surgical interventions such as muscle flap grafting or scar tissue debridement are associated with significant donor site morbidity and functional deficiency. Experimental approaches using decellularized extracellular matrix scaffolds show limited benefit in muscle recovery. Accordingly, a tissue engineering system that can restore normal skeletal muscle structure and function remains lacking for treatment of VML. Since skeletal muscle is composed generally of a bundle of parallel-aligned myofibers interspersed with blood vessels that provide blood and oxygen to the myofibers, the long-term goal of this proposal is to engineer vascularized skeletal muscle tissue constructs that mimic the native muscle and vessel structure, in order to restore muscle function after VML. The purpose of this study is to bioengineer skeletal muscle tissue composed of skeletal muscle precursor cells and vascular endothelial cells in a parallel-aligned nanofibrillar scaffold that augments cell survival, myofiber formation, and vascular perfusion recovery in a murine model of VML. Owing to the importance of vascular perfusion recovery, the scaffolds will also be engineered to release angiogenic growth factors in the form of modified mRNA (mmRNA), which obviates genomic alterations. The proposed objectives are designed to advance the understanding of how intercellular interactions with parallel-aligned nanofibrillar scaffolds, along with transient delivery of therapeutic mmRNA, can promote muscle and vascular regeneration. Accordingly, the Specific Aims are: (1) To engineer endothelialized aligned skeletal muscle composed of muscle precursor cells and endothelial cells in an aligned nanofibrillar scaffold that augments cell survival, myotube formation, and contractile function in vitro; (2) To enhance the angiogenic capacity of endothelialized and parallel- aligned engineered skeletal muscle using scaffold-mediated mmRNA delivery; and (3) To quantify the therapeutic efficacy of endothelialized and aligned engineered skeletal muscle with transient therapeutic mmRNA delivery in a murine model of VML. The proposed studies are highly significant because they seek to improve the therapeutic benefit of cell transplantation for treatment of VML, shifting away from the transplantation of acellular scaffolds to pre-formed endothelialized muscle tissue constructs with transient gene delivery for improved clinical outcomes in Veterans and other patients with VML.

容积性肌肉损失（VML）的特征是损失了相当大的一部分， 骨骼肌损伤，导致肌肉结构和功能的永久性损伤。 VML由严重的创伤性损伤引起，并且变得越来越频繁 在退伍军人由于路边爆炸，枪伤，和汽车 车祸。VML有助于长期残疾和4000亿美元的经济增长 美国每年的负担。导致VML的创伤性损伤与以下因素有关： 内源性肌肉再生和血管再生能力受损。电流 外科手术例如肌瓣移植或疤痕组织清创术， 与显著的供体部位发病率和功能缺陷相关。实验 使用脱细胞细胞的细胞外基质支架的方法显示出有限的益处， 肌肉恢复因此，可以恢复正常的组织工程系统， 骨骼肌结构和功能对于VML的治疗仍然缺乏。以来 骨骼肌通常由一束平行排列的肌纤维组成 散布着为肌纤维提供血液和氧气的血管， 这项提案的长期目标是工程化血管化骨骼肌组织 模拟天然肌肉和血管结构的结构，以恢复肌肉 VML之后的函数。 这项研究的目的是生物工程骨骼肌组织组成的 骨骼肌前体细胞和血管内皮细胞在平行排列的 增强细胞存活、肌纤维形成和血管生成的纳米原纤维支架 VML小鼠模型中的灌注恢复。由于血管的重要性 灌注恢复，支架也将被工程化以释放血管生成生长， 修饰的mRNA（mmRNA）形式的因子，其避免了基因组改变。 提出的目标旨在促进理解如何 与平行排列的纳米纤维支架的细胞间相互作用，沿着与瞬时 递送治疗性mmRNA可以促进肌肉和血管再生。 因此，具体目的是：（1）工程化内皮化排列 骨骼肌由肌肉前体细胞和内皮细胞组成， 增强细胞存活、肌管形成和收缩的纳米原纤维支架 体外功能;（2）增强内皮化和平行- 使用支架介导的mmRNA递送对齐的工程化骨骼肌;和（3） 为了量化内皮化和对齐的工程化骨骼的治疗效果， 在VML的鼠模型中用瞬时治疗性mmRNA递送肌肉。的 拟议的研究是非常重要的，因为他们寻求改善治疗， 细胞移植治疗VML的好处，从 脱细胞支架向预形成的内皮化肌肉组织的移植 用于改善退伍军人临床结果的瞬时基因递送构建体， 其他VML患者。