Notch signaling and bone fracture healing

Notch信号传导与骨折愈合

• 批准号: 8046355
• 负责人: Michael Irving Dishowitz
• 金额: $ 4.26万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8046355
• 关键词: Adopted; Animal Model; Bilateral; Biocompatible Materials; Biological; Biomaterials Research; Biomechanics; Biomedical Engineering; Bone Morphogenetic Proteins; Bone Regeneration; Bone Tissue; Bone Transplantation; Brain; Caliber; Calvaria; Cells; Clinical; Communities; Complement; Defect; Development; Exhibits; Foundations; Fracture; Fracture Healing; Gene Expression; Gene Targeting; Gold; Healed; Heart; Histology; Homeostasis; Immunohistochemistry; Implant; In Vitro; Injury; LacZ Genes; Ligands; Maintenance; Medicine; Methods; Modeling; Morbidity - disease rate; Mus; Natural regeneration; Notch Signaling Pathway; Oral; Organ; Orthopedics; Osteoblasts; Play; Population; Postoperative Pain; Proteins; Regulation; Reporter; Retina; Role; Signal Transduction; Site; Skin; Staging; Stem cells; Strategic Planning; Testing; Tibial Fractures; Time; Tissue Engineering; Tissues; Trachea; Transgenic Mice; Transgenic Organisms; Translational Research; Up-Regulation; Vascular Endothelial Growth Factors; Wild Type Mouse; Wound Healing; angiogenesis; base; bone; bone healing; craniofacial; gene therapy; healing; improved; inhibitor/antagonist; jagged1 protein; long bone; notch protein; osteoblast differentiation; osteoprogenitor cell; prevent; promoter; public health relevance; reconstruction; response to injury; scaffold; stem; success

DESCRIPTION (provided by applicant): Craniofacial and long bone defects often exhibit delayed or non-union healing. While autogenous bone grafts and bone morphogenetic proteins are widely used to treat such severe injuries, these therapies have limitations. Therefore, a clinical need persists for the development of new methods to enhance bone regeneration. The Notch signaling pathway regulates osteoprogentor proliferation and osteoblast differentiation. Additionally, Notch signaling is an essential component for wound healing in many tissues, and Notch upregulation following injury has been shown to enhance regeneration of those tissues. Therefore, the long-term objectives of this study are to show that Notch signaling also regulates regeneration of craniofacial and long bone tissue, and that upregulation of Notch signaling through a clinically-applicable tissue engineering strategy can enhance craniofacial bone regeneration. The following three specific aims complement the NIDCR's strategic plan for 2009-2013 (draft) objective 1-5, which is to 'Facilitate reconstruction and regeneration of diseased or damaged oral and craniofacial tissues and organs through biological, bioengineering and biomaterials research approaches.' For the first specific aim we will characterize Notch signaling during calvarial defect and tibial bone fracture healing. Notch reporter mice will undergo a 3 mm diameter calvarial defect or bilateral tibial fractures. Mice will be sacrificed pre-injury and at 1, 2, 5, 10, 15, 20 and 40 days post-injury for qPCR and immunohistochemistry analysis. In specific aim 2, we will determine the critical importance of Notch signaling for successful calvarial and tibial bone regeneration. Transgenic mice with Cre-regulated expression of the canonical notch inhibitor dnMAML will undergo a 3 mm calvarial defect or bilateral tibial fractures and adeno-Cre will be locally administered tp permanently inhibit Notch signaling (adeno-LacZ control). Mice will be sacrificed at 5, 10, 20 and 40 days post-injury for |JCT, histology, and biomechanical testing. For the final specific aim we will develop a Notch tissue engineering therapy to enhance calvarial bone regeneration. As a proof of concept, we will administer adeno-NICD (adeno-LacZ control) to 3 mm calvarial defects in wild type mice to transiently upregulate Notch. We will then adsorb Notch ligand, Jagged-1 onto an osteoconductive biomaterial and implant these tissue engineered constructs in 3 mm calvarial defects. The purpose of this study is to 1) investigate the role(s) of the Notch signaling pathway in craniofacial bone healing, and 2) show that Notch manipulation can clinically enhance craniofacial bone regeneration. Public Health Relevance: Craniofacial and long bone defects often exhibit delayed or non-union healing. While autogenous bone grafts and bone morphogenetic proteins are widely used to treat such severe injuries, these therapies have limitations. Therefore, a clinical need persists for the development of new methods to enhance bone regeneration. The purpose of this study is to 1) investigate the role(s) of the Notch signaling pathway in craniofacial bone healing, and 2) show that Notch manipulation can clinically enhance craniofacial bone regeneration.

描述（由申请人提供）：颅面和长骨缺损通常表现出愈合延迟或不愈合。虽然自体骨移植和骨形态发生蛋白被广泛用于治疗此类严重损伤，但这些疗法具有局限性。因此，临床仍然需要开发新方法来增强骨再生。 Notch 信号通路调节骨原细胞增殖和成骨细胞分化。此外，Notch 信号传导是许多组织伤口愈合的重要组成部分，损伤后 Notch 上调已被证明可以增强这些组织的再生。因此，本研究的长期目标是证明Notch信号也调节颅面骨和长骨组织的再生，并且通过临床适用的组织工程策略上调Notch信号可以增强颅面骨再生。以下三个具体目标补充了NIDCR 2009-2013年战略计划（草案）目标1-5，即“通过生物、生物工程和生物材料研究方法促进患病或受损的口腔和颅面组织和器官的重建和再生”。对于第一个具体目标，我们将描述颅骨缺损和胫骨骨折愈合过程中的 Notch 信号传导。 Notch报告小鼠将出现直径3毫米的颅骨缺损或双侧胫骨骨折。将在损伤前以及损伤后1、2、5、10、15、20和40天处死小鼠进行qPCR和免疫组织化学分析。在具体目标 2 中，我们将确定 Notch 信号传导对于颅骨和胫骨骨成功再生的至关重要性。具有经典Notch抑制剂dnMAML的Cre调节表达的转基因小鼠将经历3毫米颅骨缺损或双侧胫骨骨折，并且将局部施用腺-Cre以永久抑制Notch信号传导（腺-LacZ对照）。将在损伤后 5、10、20 和 40 天处死小鼠以进行 JCT、组织学和生物力学测试。为了最终的具体目标，我们将开发一种 Notch 组织工程疗法来增强颅骨再生。作为概念证明，我们将对野生型小鼠的 3 mm 颅骨缺损施用腺-NICD（腺-LacZ 对照）以暂时上调 Notch。然后，我们将 Notch 配体 Jagged-1 吸附到骨传导生物材料上，并将这些组织工程结构植入 3 毫米颅骨缺损中。本研究的目的是 1) 研究 Notch 信号通路在颅面骨愈合中的作用，2) 表明 Notch 操作可以在临床上增强颅面骨再生。 公共健康相关性：颅面和长骨缺损通常表现出愈合延迟或不愈合。虽然自体骨移植和骨形态发生蛋白被广泛用于治疗此类严重损伤，但这些疗法具有局限性。因此，临床仍然需要开发新方法来增强骨再生。本研究的目的是 1) 研究 Notch 信号通路在颅面骨愈合中的作用，2) 表明 Notch 操作可以在临床上增强颅面骨再生。