Engineering Innervated Muscle-Tendon Constructs for Tissue Regeneration

工程神经支配的肌肉肌腱结构用于组织再生

• 批准号: 7847858
• 负责人: LISA M LARKIN
• 金额: $ 8.62万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-10 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7847858
• 关键词: 3-Dimensional; Acetylcholine; Address; Adult; Advanced Development; Animals; Biochemical; Biology; Bioreactors; Caliber; Cardiovascular system; Cell membrane; Characteristics; Coculture Techniques; Collagen; Complex; Development; Electric Stimulation; Engineering; Environment; Fiber; Glycolates; Growth; Hindlimb; Implant; In Vitro; Infiltration; Injury; Isometric Exercise; Lead; Length; Limb structure; Longitudinal Studies; Mechanics; Medicine; Methodology; Muscle; Muscle Cells; Muscle Contraction; Muscle Fibers; Musculoskeletal; Myosin Heavy Chains; Neonatal; Nerve; Nerve Growth Factors; Nerve Tissue; Neuromuscular Junction; Neurons; Phenotype; Physical environment; Plasmids; Polymers; Production; Protein Isoforms; Proteins; Rattus; Recruitment Activity; Silicones; Simulate; Skeletal Muscle; Spinal Cord; Stress; Structure; Study models; System; Tail; Tendon structure; Testing; Time; Tissue Engineering; Tissues; Transfection; Work; alpha Bungarotoxin; base; design; end-stage organ failure; fetal; functional restoration; implantation; in vitro activity; in vivo; migration; muscle engineering; muscle pharmacology; nerve supply; neurotrophic factor; relating to nervous system; response; sciatic nerve; three dimensional structure; tissue regeneration

DESCRIPTION (provided by applicant): End-stage organ failure or tissue loss is one of the most devastating and costly problems in medicine. The creation of engineered musculoskeletal tissue with functional myotendinous (MTJ) and neuromuscular (NMJ) junctions will not only restore the function of complex tissues such as muscle, tendon, and nerve following traumatic injury, but can also be used as a model for studying developmental muscle biology and muscle pharmacology. We have demonstrated that the co-culture of fetal nerve tissue with fetal or engineered tendon and engineered muscle tissue produces constructs with viable muscle-tendon interfaces that remain intact during force production, and viable neuromuscular junctions that advance the phenotype of the muscle tissue within the construct [1, 2]. The tissues formed express neonatal structures and do not substantially advance functionally or phenotypically in the absence of electrical or mechanical loading. The purpose of this study is to design, fabricate, and evaluate the structural and contractile characteristics of three-dimensional (3-D) engineered tissues containing myotendinous junctions (MTJ) and neuromuscular junctions (NMJ), two of the principal tissue interfaces required for a functional musculoskeletal construct. We propose to study the long-term viability of these constructs; to increase the expression of neurotrophic proteins like glial derived neurotrophic factor (GDNF) and introduce synthetic, engineered, and tissue based conduits to enhance innervation of the muscle constructs; to use bioreactors to place the nerve-tendon- muscle constructs into physical environments which simulate the stress, strain, and contractile activity resembling the mechanical milieu found in hind limb muscles in vivo; and to implant the constructs in vivo to surround the construct with the actual mechanical and biochemical environment of a hindlimb. Our working hypothesis is that an increase in the number of NMJs in conjunction with applied electrical activity in vitro will lead to a more advanced phenotype in the muscle and tendon, increased force production in the engineered muscles, and stronger and more developed tissue interfaces. In vivo implantation of in vitro engineered constructs will further advance the phenotype and functionality of all tissues and tissue interfaces. This project will address the following specific aims: Specific Aim 1. To design and fabricate nerve-tendon-muscle constructs with functional MTJs and NMJs and evaluate both the structural and functional integrity of the constructs at three time points of development. Specific Aim 2. To develop a conduit system to direct the growth of multiple neural extensions towards the muscle construct thus increasing the innervation ratio of the construct. Specific Aim 3. To evaluate the effect of muscle contraction in response to electrical stimulation on the structure and function of the nerve-tendon-muscle construct. Specific Aim 4. To evaluate the effect of an in vivo implantation into a host animal on the structure and function of the nerve-muscle tendon construct at four weeks.

描述（由申请人提供）：终末期器官衰竭或组织损失是医学上最具破坏性和代价最高的问题之一。具有功能性肌腱（MTJ）和神经肌肉（NMJ）连接的工程化肌肉骨骼组织的创建不仅将恢复创伤性损伤后的复杂组织如肌肉、肌腱和神经的功能，而且还可以用作研究发育肌肉生物学和肌肉药理学的模型。我们已经证明，胎儿神经组织与胎儿或工程化肌腱和工程化肌肉组织的共培养产生了具有在力产生期间保持完整的可行肌肉-肌腱界面的构建体，以及促进构建体内肌肉组织表型的可行神经肌肉接头[1，2]。所形成的组织表达新生结构，并且在没有电或机械负载的情况下在功能上或表型上基本上不前进。本研究的目的是设计，制造，并评估三维（3-D）工程组织的结构和收缩特性，包含肌腱接头（MTJ）和神经肌肉接头（NMJ），两个主要的组织界面所需的功能性肌肉骨骼结构。我们建议研究这些结构的长期生存能力;增加神经营养蛋白如胶质源性神经营养因子（GDNF）的表达，并引入合成的、工程化的和基于组织的导管来增强肌肉结构的神经支配;使用生物反应器将神经-肌腱-肌肉结构置于模拟应力，应变，和类似于在体内后肢肌肉中发现的机械环境的收缩活性;以及将所述构建体植入体内以用后肢的实际机械和生物化学环境包围所述构建体。我们的工作假设是，NMJ数量的增加与体外应用的电活动相结合，将导致肌肉和肌腱中更高级的表型，工程肌肉中的力产生增加，以及更强大和更发达的组织界面。体外工程构建体的体内植入将进一步促进所有组织和组织界面的表型和功能。该项目将针对以下具体目标：具体目标1。设计和制造具有功能性MTJ和NMJ的神经-肌腱-肌肉结构，并在三个开发时间点评估结构和功能的完整性。具体目标2。开发一种导管系统，以引导多个神经延伸向肌肉结构生长，从而增加结构的神经支配率。具体目标3。评价电刺激引起的肌肉收缩对神经-肌腱-肌肉结构和功能的影响。具体目标4。评价体内植入宿主动物后4周对神经-肌腱结构和功能的影响。