Aligned Tendon or Ligament Derived Matrix Nanoscaffolds for Rotator Cuff Repair

用于肩袖修复的对齐肌腱或韧带衍生基质纳米支架

• 批准号: 8251126
• 负责人: Dianne Little
• 金额: $ 9.8万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-04 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8251126
• 关键词: Adipose tissue; Animal Model; Anisotropy; Architecture; Award; Basic Science; Biochemical; Biological Assay; Biomechanics; Biomedical Engineering; Cells; Chronic; Clinical; Collagen; Collagen Fiber; Collection; Confocal Microscopy; Consultations; DNA; Data; Deposition; Development; Devices; Disease; Doctor of Philosophy; Educational process of instructing; Environment; Equipment; Extracellular Matrix; Failure; Fiber; Fourier Transform; Gastrointestinal Physiology; Gene Expression; Goals; Human; Image; In Vitro; Infiltration; Inflammation; Joint Capsule; Laboratory Research; Left; Ligaments; Manuscripts; Measures; Mechanics; Mentorship; Modification; Muscle; Nuclear; Operative Surgical Procedures; Orthopedics; Phenotype; Porosity; Positioning Attribute; Process; Property; Proteoglycan; Publishing; Reporting; Research; Research Personnel; Research Project Grants; Research Training; Resistance; Rodent Model; Rotator Cuff; Scanning Electron Microscopy; Shapes; Shoulder; Solid; Solutions; Stem cells; Stress; Stress Tests; Structure; Surface; Surgeon; Surgical sutures; Tenascin; Tendon structure; Testing; Time; Tissue Engineering; Tissues; Training; Work; aqueous; biglycan; biomaterial compatibility; biomaterial development; bone; career; career development; cell motility; decorin; electric field; experience; in vivo; injury and repair; innovation; meetings; nanofiber; novel; novel strategies; polysulfated glycosaminoglycan; post-doctoral training; repaired; responsible research conduct; scaffold; scleraxis; skills; standard care; standard of care; statistics; success

DESCRIPTION (provided by applicant): Candidate: The candidate is a board-certified veterinary surgeon with extensive research training in animal models and basic science who transitioned after earning a PhD in gastrointestinal physiology to postdoctoral training in biomedical engineering. Her immediate career development goals in the current proposal will be in biomechanics and tissue engineering and will allow expansion of current skills in mechanical testing and stress-strain analysis, biomaterial development, confocal microscopy, image and data post-processing and fast Fourier transform analysis, skills essential to the success of the research project and long term career goals as an independent investigator in the field of tendon and ligament tissue engineering. The candidate plans R03 submission in year 2 to evaluate biocompatibility of the proposed scaffolds in rodent models, and then an R01 submission in year 4 to evaluate the proposed scaffolds in a large animal model of chronic rotator cuff tear, and expects to publish three manuscripts resulting from this initial work. Additional didactic training in tissue engineering, statistics and responsible conduct of research will be obtained during the award period. Environment: The mentorship of Dr. Farshid Guilak PhD and co-mentorship of Dr. David Ruch MD, combined with the degree of institutional commitment will provide this candidate with the support needed to transition successfully into an independent research career. External consultation from experts in rotator cuff disease, research and repair will provide added expertise. All the equipment and facilities needed by the candidate for successful completion of this proposal are already available in the Orthopaedic Research Laboratories in the Department of Surgery, and the candidate has 100% protected time, with no administrative, clinical or teaching responsibilities to allow for the successful completion of this project. Research: The rotator cuff is a composite of joint capsule, tendons and ligaments that is critical for stability and function of the shoulder. There are over 300,000 rotator cuff injuries repaired annually in the US, at an annual burden in surgical expenses alone of $3-4 billion. Suture repair is the clinical standard of care but re- tear rates of 34-94% have been reported. Extracellular matrix (ECM) scaffolds are used for augmentation of rotator cuff repair because they contain bioactive molecules that stimulate cell migration, proliferation and ECM synthesis. All of the currently available ECM scaffold patches used to augment suture repair have limitations and none mimic the interdigitating multi-layered structure of the normal rotator cuff which has layers of collagen aligned at 45 degrees to each other to allow efficient load transfer and distribution, suture retention and resistance to failure. Therefore, there is a need for a rotator cuff patch that meets all of the deficiencies of currently available devices, while retaining the benefits of using ECM. The long term goal of the proposed research is to develop a tendon or ligament derived-extracellular matrix (TLDM) patch for augmentation of rotator cuff repair or as a tendon substitute in irreparable rotator cuff tears while allowing the PI to establish an independent research career in tissue engineering of tendon and ligament. Electrospun nanofibers are attractive for tissue engineering of a rotator cuff patch because the fibers can be aligned to mimic the natural architecture and biomechanical properties of native tissue. Electrospinning into an aqueous solution and collection of multiple layers of scaffold (hydrospinning) increases porosity and further enhances cell infiltration compared to more traditional nanoscaffolds deposited as a single layer onto a solid ground plate. We have shown that a novel TLDM enhances proliferation and early differentiation of human adipose stem cells (hASCs) towards a tendon phenotype and that TLDM can be used to produce hydrospun nanofiber scaffolds. We have developed a novel strategy to align adjacent layers of hydrospun nanofibers in different directions by modification of the shape of the electric field. The PI is well prepared to meet the scientific and training goals of this application through the research experience already acquired in the proposed research and mentorship environment, and through the mentorship and continued development in this environment covered in this proposal. The overall hypothesis is that aligned TLDM scaffolds will show anisotropic ECM maturation compared to non-aligned scaffolds, and scaffolds with layers aligned at 45 degrees to each other (45-Offset) will demonstrate increased mechanical properties over a range of load directions than TLDM scaffolds with all layers aligned similarly (0- Offset) or non-aligned TLDM scaffolds. Use of 45-Offset layers of hydrospun fibers represents an innovative approach to promote early ECM synthesis, alignment and maturation in a multi-layered RC tendon TLDM patch. We will test the following hypotheses: Hypothesis 1: Scaffold fiber alignment and mechanical anisotropy will be greatest in 0-Offset scaffolds followed by 45-Offset scaffolds and finally non-aligned TLDM scaffolds. Hypothesis 2: 45-Offset and 0-Offset scaffolds will show increased cell and ECM alignment and maturation when seeded with hASCs, compared to non-aligned scaffolds. We expect that 45-Offset aligned TLDM scaffolds will permit earlier and more aligned ECM synthesis and maturation compared to non-aligned scaffolds and reduced mechanical anisotropy compared to 0-Offset aligned scaffolds.

描述（由申请人提供）：候选人：候选人是一名经过董事会认证的兽医外科医生，对动物模型和基础科学进行了广泛的研究培训，他们在获得胃肠道生理学博士学位后转变为生物医学工程的博士后培训。 Her immediate career development goals in the current proposal will be in biomechanics and tissue engineering and will allow expansion of current skills in mechanical testing and stress-strain analysis, biomaterial development, confocal microscopy, image and data post-processing and fast Fourier transform analysis, skills essential to the success of the research project and long term career goals as an independent investigator in the field of tendon and ligament tissue engineering.候选人计划在第2年内提交R03提交，以评估啮齿动物模型中提议的脚手架的生物相容性，然后在第4年进行R01提交，以评估在大型慢性肩袖撕裂的大动物模型中提出的脚手架，并期望由这项初始工作产生三个手稿。在奖励期间将获得组织工程，统计和负责任的研究的其他教学培训。环境：Farshid Guilak博士博士的指导和大卫·鲁奇（David Ruch）博士的教会，加上机构承诺的程度，将为这位候选人提供成功过渡到独立研究职业所需的支持。肩袖疾病，研究和维修专家的外部咨询将提供更多的专业知识。候选人成功完成本提案所需的所有设备和设施都已在外科部的骨科研究实验室中提供，候选人有100％受保护的时间，没有行政，临床或教学责任，无法成功完成该项目。研究：肩袖是关节胶囊，肌腱和韧带的复合物，对于肩膀的稳定性和功能至关重要。每年在美国每年进行300,000多个肩袖伤害，仅手术费用为3-4亿美元。缝合线修复是临床护理标准，但据报道，撕裂率为34-94％。细胞外基质（ECM）支架用于增强肩袖修复，因为它们包含刺激细胞迁移，增殖和ECM合成的生物活性分子。用于增强缝合线修复的所有当前可用的ECM脚手架贴片都有局限性，并且没有模仿正常肩袖的互相结构的结构，该结构的胶原蛋白层以45度排列在45度中，以允许有效的载荷转移和分布，Suture retrestion和分布，抵抗力和对失败的抵抗力。因此，需要一个符合当前可用设备的所有缺陷的肩袖贴片，同时保留了使用ECM的好处。拟议的研究的长期目标是开发肌腱或韧带衍生的 - 细胞基质（TLDM）贴片，以增强肩袖修复的增强或作为无法弥补的肩袖撕裂的肌腱替代品，同时允许PI在肌腱和韧带的组织工程中建立独立的研究职业。电纺纳米纤维对肩袖贴片的组织工程具有吸引力，因为纤维可以对齐以模仿天然组织的天然结构和生物力学特性。与将单个层作为单层沉积在实心接地板上的更传统的纳米镜相比，静电纺入水溶液和多层脚手架（水纺）的集合会增加孔隙率，并进一步增强细胞浸润。我们已经表明，一种新型的TLDM增强了人脂肪干细胞（HASC）向肌腱表型的增殖和早期分化，并且可以使用TLDM来产生水传物纳米纤维支架。我们已经开发了一种新的策略，可以通过修改电场的形状来对齐在不同方向上的氢传纳米纤维层。 PI已准备好通过拟议的研究和指导环境中已经获得的研究经验以及通过本提案中涵盖的这种环境中的指导和持续发展来实现该应用程序的科学和培训目标。总体假设是，与非对齐的脚手架相比，对齐的TLDM支架将显示各向异性ECM成熟，并且在45度以45度排列的层（45下降）将表明机械性能增加的机械性能增加了与所有layers and ofligned and ofligned（0）相似的机械性能（45下），将表明机械性能增加（0）。脚手架。在多层RC肌腱TLDM贴片中，使用45个偏移层的液压纤维代表了一种促进ECM合成，对齐和成熟的创新方法。我们将检验以下假设：假设1：支架纤维比对和机械各向异性在0折叠支架中将是最大的，然后是45个偏移的支架，最后是非对准的TLDM支架。与非对齐的支架相比，假设2：45偏移和0偏移支架将显示出增加的细胞和ECM比对和成熟。我们预计，与非对准的支架相比，与0偏移的支架相比，与非对齐的支架相比，与0偏移的支架相比，与非对齐的支架相比，与0偏移的机械各向异性相比，ECM的合成和成熟度更早，比对齐的ECM合成和成熟。