Extracellular Matrix Impacts Angiogenesis and Growth Plate Repair

细胞外基质影响血管生成和生长板修复

• 批准号: 10668056
• 负责人: Melissa Krebs
• 金额: $ 16.89万
• 依托单位: COLORADO SCHOOL OF MINES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10668056
• 关键词: Address; Affect; Alginates; Angiogenesis Inhibition; Angiogenesis Inhibitors; Angiogenic Peptides; Animal Model; Antibodies; Area; Biocompatible Materials; Bone Growth; Cartilage; Child; Childhood; Chitosan; Chondrogenesis; Clinic; Clinical; Cues; Deformity; Development; Endothelial Cells; Environment; Epiphysial cartilage; Extracellular Matrix; Fracture; Goals; Growth; Histologic; Hyaluronic Acid; Hydrogels; Impairment; In Vitro; Injectable; Injury; Link; Modeling; Modification; Molecular Weight; Operative Surgical Procedures; Osteogenesis; Pathway interactions; Peptides; Physiological; Play; Prevention; Process; Rattus; Research; Role; Signal Pathway; Signal Transduction; Site; Skeleton; System; Technology; Tissues; Translating; Translations; Vascular Endothelial Growth Factors; Work; angiogenesis; biomaterial development; bone; cartilaginous; cell behavior; cost; disability; healing; in vivo; injured; innovation; long bone; microCT; novel; pediatric patients; prevent; protein aminoacid sequence; regenerative; regenerative therapy; repaired; response; tissue repair; transcriptome sequencing; treatment strategy

PROJECT SUMMARY Growth plate injuries, which account for 30% of all pediatric fractures, can impair bone growth and even halt it completely. For children who are still growing, these injuries can be devastating. The growth plate (or physis) is a cartilage region found at the end of all long bones in children and is responsible for longitudinal bone growth. It is a weak area of the developing skeleton and prone to injury. Once damaged, cartilage tissue within the growth plate can be replaced by unwanted bony tissue, forming a “bony bar”, which can lead to angular deformities or complete growth arrest. Pediatric patients who sustain these injuries may require multiple surgical interventions during childhood. Innovative treatment strategies that prevent initial bony bar formation, thus avoiding growth deformities and potential lifelong disability, are critically needed. The goal of this project is to develop clinically useful treatment strategies for growth plate injuries that prevent bony bar formation and associated growth problems. One approach is to target mechanisms responsible for unwanted bony repair tissue, which include angiogenic signaling pathways. These pathways are regulated at many levels and can be modulated by insoluble cues such as extracellular matrix factors. The modulation of these cues via a material-only system could provide significant benefit to ultimate translation of such a regenerative therapy. Our hypothesis is that targeted disruption of angiogenic signaling cascades after growth plate injury through insoluble cues such as extracellular matrix factors will inhibit angiogenesis and completely prevent bony bar formation. We will examine this with the following 2 aims: AIM 1: To quantify the impact of hyaluronic acid (HA) on the angiogenic response that occurs after growth plate injury. As HA is known to be important in angiogenic signaling, but as its effect can be varied in different physiologic settings and as its impact in the growth plate after injury has not been studied, here we will investigate varying molecular weights of HA. Angiogenesis and bony bar prevention will be evaluated in our rat model of growth plate injury using bulk RNA-seq, microCT, immunostaining and histological assessment. AIM 2: To quantify the impact of specific peptide sequences on the angiogenic response that occurs after growth plate injury. Here 4 different peptides with established inhibitory effects on angiogenesis and osteogenesis will be covalently linked to our alginate hydrogels to study their influence on cell behavior. This Aim will quantify the impact of these peptides on angiogenesis, osteogenesis, and chondrogenesis in vitro and on the angiogenesis and osteogenesis that occurs in vivo in a rat growth plate injury model, as quantified using bulk RNAseq, microCT, immunostaining, and histological assessment. This project will provide important information about the impact of extracellular matrix cues in de novo growth plate injury healing and bony bar formation, supporting the development of novel biomaterial-based approaches to preventing bony bar formation. Ultimately, we will translate the technology to larger animal models of growth plate injuries, and eventually into the clinic. This research will help address a critical unmet clinical need for children suffering from growth plate injuries.

项目摘要 生长板损伤，占所有儿科骨折的30%，可损害骨骼生长，甚至停止它 彻底对于正在成长的孩子来说，这些伤害可能是毁灭性的。生长板（或骺板）是 儿童所有长骨末端的软骨区域，负责纵向骨骼生长。 它是骨骼发育中的薄弱部位，容易受伤。一旦受损，软骨组织内的生长 接骨板可能被不需要的骨组织替代，形成“骨棒”，这可能导致成角畸形或 完全停止生长这些损伤的儿科患者可能需要多次手术干预 在童年时期。创新的治疗策略，防止最初的骨棒形成，从而避免生长 残疾和潜在的终身残疾，是迫切需要的。该项目的目标是临床开发 用于防止骨棒形成和相关生长的生长板损伤的有效治疗策略 问题一种方法是靶向负责不需要的骨修复组织的机制，包括 血管生成信号通路。这些途径在许多水平上受到调节，并且可以通过不溶性的 细胞外基质因子等线索。通过一个纯物质系统对这些线索的调节可以提供 这对这种再生疗法的最终转化有显著益处。我们的假设是， 生长板损伤后，通过细胞外基质等不溶性线索， 因子将抑制血管生成并完全阻止骨棒形成。我们将用 以下2个目的：目的1：量化透明质酸（HA）对血管生成反应的影响， 生长板损伤后已知HA在血管生成信号传导中是重要的，但由于其作用可以变化， 在不同的生理环境中，由于其对损伤后生长板的影响尚未研究， 将研究HA的不同分子量。血管生成和骨杆预防将在我们的 使用批量RNA-seq、microCT、免疫染色和组织学评估的大鼠生长板损伤模型。目的 2：量化特定肽序列对生长后发生的血管生成反应的影响 钢板损伤在这里，4种不同的肽具有确定的抑制血管生成和骨生成的作用， 共价连接到我们的藻酸盐水凝胶，以研究它们对细胞行为的影响。这一目标将量化 这些肽对体外血管生成、骨生成和软骨生成的影响以及对血管生成的影响 和在大鼠生长板损伤模型中体内发生的骨生成，如使用批量RNAseq，microCT， 免疫染色和组织学评估。该项目将提供有关影响的重要信息， 细胞外基质在新生生长板损伤愈合和骨棒形成中的作用，支持 开发新的生物材料为基础的方法，以防止骨杆形成。最终，我们将 将这项技术转化为生长板损伤的大型动物模型，并最终应用于临床。这 研究将有助于解决生长板损伤儿童的一个关键的未满足的临床需求。