Muscle-based tissue engineering to improve bone healing

基于肌肉的组织工程改善骨愈合

基本信息

批准号：
7676899
负责人：
Johnny Huard
金额：
$ 45.45万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-08-01 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7676899
关键词：
Address Adipose tissue Adult Affect Age Angiogenic Factor Angiogenic Proteins Animal Model Applications Grants Arts Autologous BMP2 gene BMP4 Biological Assay Biology Biopsy Blood Circulation Blood Vessels Bone Injury Bone Marrow Bone Regeneration Bone Tissue CD34 gene Calvaria Cell Count Cell Differentiation process Cell Proliferation Cell Separation Cells Chemicals Clinical Treatment Data Defect Desmin Development Embryo Employee Strikes Endothelial Cells Engineering Enhancers Exhibits Female Fracture Funding Future Galactosidase Gel Genetic Engineering Germ Cells Goals Green Fluorescent Proteins Half-Life Healed Heart Human Human Biology Image Implant In Vitro Inferior Injectable Injection of therapeutic agent Injury International Kinetics Laboratories LacZ Genes Lead Life Light MAP Kinase Gene MAPK11 gene MAPK14 gene Mechanical Stimulation Mechanics Mediating Mesenchymal Stem Cells Methods Microscopic Modeling Modification Monitor Multipotent Stem Cells Mus Muscle Muscle Fibers Muscle satellite cell Musculoskeletal Musculoskeletal System Myoblasts NIH 3T3 Cells National Institute of Dental and Craniofacial Research Natural regeneration Nature Orthopedic Surgery procedures Osteocalcin Osteogenesis Paper Pathway interactions Patients Pericytes Play Population Process Proteins Protocols documentation Recruitment Activity Regenerative Medicine Reporting Research Research Personnel Retroviral Vector Retroviridae Reverse Transcriptase Polymerase Chain Reaction Role SCID Mice Sex Characteristics Signal Pathway Signal Transduction Site Skeletal Muscle Skeleton Source Staining method Stains Stem cells Stromal Cells Surgeon System Techniques Technology Testing Time Tissue Engineering Tissues Transgenic Mice Transplantation Vascular Endothelial Cell Vascular Endothelial Growth Factor Receptor Vascular Endothelial Growth Factors Vascular Endothelium Vascular blood supply abstracting adult stem cell age effect age related angiogenesis base bone bone cell bone healing bone morphogenetic protein 2 cellular transduction clinical application craniofacial cranium design genetic manipulation healing human female human tissue implantation improved in vivo injured long bone male meetings member mineralization mouse model muscle regeneration new therapeutic target older patient osteogenic osteogenic protein osteopontin postnatal progenitor promoter prospective protein expression recombinase regenerative repaired research study scaffold sex stem cell differentiation

项目摘要

Incomplete healing of bone defects ih the craniofacia.l skeleton is common. Osteogenic proteins, including bone morphogenetic protein 2 and 4 (BMP2, BMP4), promote healing in bone defects, but the proteins' short half-lives and rapid clearance by the bloodstream limit their utility. The main goal of our initial R01 and the first competitive renewal project was the development of tissue engineering approach'es, based on muscle-derived stem cells (MDSCs), to efficiently deliver osteogenic proteins and improve craniofacial bone healing. In brief, during this funding period, we demonstrated that MDSCs genetically engineered to express BMP2 and BMp4 differentiate toward an osteogenic lineage and can improve bone healing in calvarial and long bone defects. We also found that concomitant expression Of vasCular endothelial growth factor (VEGF) improves the bone healing observed after implantation Of BMP2 and 13MP4 expressing MDSCs. Additionally, we have demonstrated that donor sex influences the in vitro osteogenic potential and in vivo bone regeneration potential of murine MDSCs and also identified wa.ys, such as genetic engineering and manipulation of the BMP signaling pathways, to improve the osteogenic potential of MDSCs. Finally, we have isolated the human equivalents of the murine MDSCs and determine their osteogenic potential in vitro. We would like to thank NIDCR for their support during the prior funding period. We met and exceeded all the key objectives in the original R01 application as well as the first competitive renewal, and our results formed the basis for 33 + papers and 117+ abstracts. This DE013420 second cornpetitive renewal application outlines experiments designed to extend these initial findings and lead to. possible future clinical applications of MDSCs to improve bone healing. We will focus this second competitive renewal on human equivalents to murine MDSCs and optimization of their use for bone regeneration. Since after implantation of MDSCs into injured musculoskeletal tissues, including bone, the repair process is often mediated by chemoattraction of host cells, we plan to determine the influence of host cells(especially blood vessel wall progenitors) chemoattraCted by donor human cells during the bone healing process (Aim 1). We plan to examine the effect of age and sexotdonor patient on the number and osteogenic potential of hMDCs derived from that patient. We then will investigate ways to optimize bone formation and healing by using hMDC-based tissue engineering, including the modulation of BMP signaling through inhibition of ERK1/2, pj8 MAPK and PI3K pathwa.ys and mechanical stim'ulation of hMDCs prior to implantation (Aim 2). The proposed experiments will provide important information regarding the basic biology of hMDCs and their use for bone healing and further the development of clinical treatments for osseous deficiencies.

骨缺损的愈合不完全，骨骼骨骼很常见。成骨蛋白，包括骨形态发生蛋白2和4（BMP2，BMP4），可促进骨缺损的愈合，但是蛋白质的短半衰期和血液流的快速清除限制了其效用。我们最初的R01和第一个竞争性更新项目的主要目标是基于肌肉衍生的干细胞（MDSC）的组织工程方法的开发，以有效地提供成骨蛋白并改善颅骨骨愈合。简而言之，在此资助期间，我们证明了MDSC在基因上设计以表达BMP2和BMP4的依靠成骨谱系，并且可以改善钙钙和长骨缺陷的骨骼愈合。我们还发现，血管内皮生长因子（VEGF）的同时表达可改善植入BMP2和13MP4表达MDSC后观察到的骨骼愈合。此外，我们已经证明，供体性别会影响鼠MDSC的体外成骨潜力和体内骨再生潜力，并确定了WA.YS（例如基因工程和BMP信号通路的操纵），以提高MDSC的成骨潜力。最后，我们分离了鼠MDSC的人体当量，并在体外确定其成骨潜力。我们要感谢NIDCR在以前的资助期间的支持。我们实现了原始R01应用程序中的所有关键目标以及第一个竞争续订，我们的结果构成了33 +论文和117+摘要的基础。此DE013420第二次玉米式更新应用程序概述了旨在扩展这些初始发现并导致的实验。 MDSC的未来临床应用可能改善骨骼愈合。我们将把第二个竞争性更新重点放在鼠类MDSC和优化其用于骨再生的使用。由于将MDSC植入包括骨骼在内的受伤的肌肉骨骼组织（包括骨骼）之后，通常是通过宿主细胞的趋化过程介导的，因此我们计划确定托管细胞（尤其是血管壁祖细胞）在骨骼愈合过程中由供体人类细胞介绍的host宿主细胞（尤其是血管壁祖细胞）的影响（AIM 1）。我们计划检查年龄和SEXOTDONOR患者对来自该患者的HMDC的数量和成骨潜力的影响。然后，我们将调查方法通过使用基于HMDC的组织工程来优化骨形成和愈合，包括通过抑制ERK1/2，PJ8 MAPK和PI3K PATHWA.YS来调节BMP信号传导，以及在植入之前对HMDC的机械刺激（AIM 2）。提出的实验将提供有关HMDC的基本生物学及其用于骨骼愈合的重要信息，并进一步发展骨缺陷的临床治疗方法。