Tissue Engineered Repair of Cranial Facial Muscle

颅面肌的组织工程修复

• 批准号: 8604631
• 负责人: Emily Lynn Ongstad
• 金额: $ 4.3万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-20 至 2015-07-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8604631
• 关键词: 3-Dimensional; Abdominal Muscles; Address; Animals; Atrophic; Autologous; Binding; Biomedical Engineering; CTF1 gene; Cell Culture Techniques; Cell Line; Cephalic; Cicatrix; Clinical; Clinical Research; Collagen; Connexin 43; Cornea; Cutaneous; DNA Sequence Rearrangement; Data; Defect; Deformity; Deposition; Development; Differentiation Inhibitor; Disease; Face; Facial Muscles; Family suidae; Fellowship; Fibrosis; Food; Funding; Future; Genes; Goals; Growth; Head and Neck Muscle; Heart; Implant; In Vitro; Inflammation; Injury; Joints; Lead; Malignant Neoplasms; Mentors; Methods; Modeling; Muscle; Muscular Atrophy; Myofibroblast; National Institute of Dental and Craniofacial Research; Natural regeneration; Operative Surgical Procedures; Outcome; Patients; Peptides; Protein Isoforms; Proteins; Publishing; Rattus; Reporting; Research; Rodent; Role; Signal Transduction; Silicones; Skeletal Muscle; Skin; Smiling; Stem cells; Structure; Surgeon; Techniques; Testing; Tissue Engineering; Tissues; Trauma; Tubulin; Universities; Work; Wound Healing; capsule; fetal; functional restoration; improved; in vivo Model; injured; member; mimetics; monolayer; programs; public health relevance; regenerative; repaired; response; satellite cell; skeletal muscle differentiation; small hairpin RNA; stem cell population; tissue reconstruction; tissue repair

DESCRIPTION (provided by applicant): The goal of my Ruth L. Kirschstein Fellowship application is to develop a tissue-engineered skeletal muscle construct for repair of cranial facia defects. We propose to investigate the mechanism by which the Connexin43 (Cx43) carboxyl terminal (CT) mimetic peptide ¿CT1 modulates the wound healing response and promotes regenerative repair of injured muscle by a tissue-engineered construct. Cranial facial muscle loss due to disease or trauma can impair the ability to chew food or smile. Current surgical repair involves rearrangement of skeletal muscle, though these approaches often fail when the muscle atrophies or results in fibrosis. Attempts at regenerating skeletal muscle lead to some degree of new muscle formation, though fibrosis is a persistent problem. In skin, promiscuous TGF¿ signaling leads to increased collagen deposition and scarring. In skeletal muscle, TGF¿ is a potent inhibitor of differentiation. Cx43 has been shown to activate TGF¿ signaling by competing with smad2 for ¿-tubulin binding. Our joint group has shown that ¿CT1 reduces inflammation and scarring in skin wounds, and reduces myofibroblast differentiation and preserves underlying muscle in rodent and porcine models of implant contraction capsules - all effects which appear to be antagonistic of TGF¿ signaling. In preliminary data we have determined that ¿CT1 reduces collagen deposition in a model of rat skeletal muscle repair by a 3D tissue-engineered construct containing autologous satellite cells (SCs). This application tests the hypothesis that targeting Cx43 function via ¿CT1 or gene knockdown promotes skeletal muscle differentiation and reduces fibrosis in tissue engineered skeletal muscle constructs through reduced downstream TGF¿ signaling. This hypothesis will be tested in three aims: In the first aim, we will conduct a cell culture study on mechanism to test the prediction that ¿CT1 or Cx43 shRNA reduces proliferation and promotes differentiation of the C2C12 myogenic cell line in vitro via reduced downstream TGF¿ signaling. In the second aim, we will conduct a study on mechanism in 2D monolayers and in an existing 3D collagen tissue-engineered construct using primary rat satellite cells (rSCs) to test the prediction that ¿CT1 or Cx43 shRNA reduces downstream TGF¿ signaling and results in reduced proliferation and increased differentiation in vitro. In the third aim, we will conduct an animal study to test the prediction that targeting Cx43 with ¿CT1 or Cx43 shRNA in the 3D tissue-engineered construct containing rSCs promotes regenerative integration of injured muscle in a cranial facial defect model in vivo. This work will be carried out under NIDCR funded RO-1 project DE019355-01A2 led jointly by my research mentor Dr. Robert Gourdie, and a member of my dissertation committee Dr. Michael Yost. The other members of my committee include Drs. Martine LaBerge (Chair, Bioengineering Department, Clemson University) and Richard Swaja (Director, Clemson-MUSC Bioengineering Program). This work will contribute to the clinical need of providing an effective means of regenerating structure and function following significant injury to cranial facial skeletal muscle.

描述（由申请人提供）：我的露丝L的目标。Kirschstein奖学金申请旨在开发一种组织工程骨骼肌构建体，用于修复颅面缺陷。我们建议通过组织工程构建来研究连接蛋白43（Cx43）羧基末端（CT）模拟肽<$CT1调节创伤愈合反应和促进损伤肌肉再生修复的机制。由于疾病或创伤导致的颅面部肌肉损失会损害咀嚼食物或微笑的能力。目前的手术修复涉及骨骼肌的重排，尽管当肌肉萎缩或导致纤维化时，这些方法通常会失败。再生骨骼肌的尝试导致一定程度的新肌肉形成，尽管纤维化是一个持续存在的问题。在皮肤中，混杂的TGF β信号导致胶原沉积增加和瘢痕形成。在骨骼肌中，TGF ²是一种有效的分化抑制剂。Cx43已被证明通过与smad 2竞争β-微管蛋白结合来激活TGF β信号传导。我们的联合小组已经表明，<$CT1可以减少皮肤伤口的炎症和瘢痕形成，减少肌成纤维细胞分化，并在植入收缩胶囊的啮齿动物和猪模型中保留潜在的肌肉-所有这些作用似乎都是TGF <$信号的拮抗作用。在初步数据中，我们已经确定，在大鼠骨骼肌修复模型中，通过含有自体卫星细胞（SC）的3D组织工程构建，CT 1减少了胶原沉积。本申请测试了通过CT 1或基因敲低靶向Cx43功能促进骨骼肌分化并通过减少下游TGF β信号传导减少组织工程化骨骼肌构建体中的纤维化的假设。这一假设将在三个目标中进行测试：在第一个目标中，我们将进行细胞培养研究的机制，以测试预测，<$CT1或Cx43 shRNA通过减少下游TGF <$信号转导减少体外C2 C12肌源性细胞系的增殖并促进其分化。在第二个目标中，我们将使用原代大鼠卫星细胞（rSC）在2D单层和现有的3D胶原组织工程构建体中进行机制研究，以测试预测<$CT1或Cx43 shRNA减少下游TGF <$信号传导并导致体外增殖减少和分化增加。在第三个目标中，我们将进行动物研究，以验证靶向Cx43的预测， 与CT 1或Cx43 shRNA在含有rSCs的3D组织工程构建体中促进颅面缺损模型中受损肌肉的再生整合这项工作将 在NIDCR资助的RO-1项目DE 019355 - 01 A2下进行，该项目由我的研究导师Robert Gourdie博士和我的论文委员会成员Michael Yost博士共同领导。我的委员会的其他成员包括Martine LaBerge博士（克莱姆森大学生物工程系主任）和Richard Swaja（克莱姆森-南加州大学生物工程项目主任）。这项工作将有助于提供一种有效的手段，再生结构和功能后，颅面骨骼肌严重损伤的临床需要。