Angiogenesis and Long-term Bone and Joint Allotransplant Survival

血管生成和长期骨和关节同种异体移植存活

• 批准号: 7858279
• 负责人: ALLEN T BISHOP
• 金额: $ 32.91万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7858279
• 关键词: Acute; Adult; Allogenic; Allografting; Anastomosis - action; Animals; Area; Autologous Transplantation; Basic Science; Biodegradable microsphere; Biomechanics; Blood Circulation; Blood capillaries; Blood flow; Bone remodeling; Cartilage; Cells; Chimerism; Chronic; Clinical; Cyclophilins; Defect; Dimensions; Effectiveness; Encapsulated; Excision; FGF2 gene; Face; Failure; Female; Femur; Fibroblast Growth Factor 2; Fluorochrome; Fracture; Genes; Goals; Grant; Growth Factor; Healed; Health; Histocompatibility; Histology; Housekeeping Gene; Hyaline Cartilage; Hydrogen; Immune; Immunosuppression; Immunosuppressive Agents; Implant; Infection; Joint Capsule; Joints; Knee; Knee joint; Label; Life; Ligaments; Limb Salvage; Limb structure; Maintenance; Measures; Mechanics; Methods; Modeling; Morbidity - disease rate; Musculoskeletal; Neoplasm Metastasis; Nutrient; Operative Surgical Procedures; Orthopedics; Oryctolagus cuniculus; Osteocytes; Osteogenesis; Outcome; Pharmaceutical Preparations; Pharmacotherapy; Polymerase Chain Reaction; Postoperative Period; Procedures; Property; Prosthesis; Rattus; Relative (related person); Risk; Sampling; Shapes; Solutions; Specimen; Stress Fractures; Structure; Testing; Time; Tissue Viability; Tissues; Transplantation; Transplanted tissue; Vascular Endothelial Growth Factors; Work; Y Chromosome; allotransplant; angiogenesis; bone; bone loss; bone morphogenic protein; capillary; density; functional outcomes; healing; immunoregulation; implantation; improved; joint function; kinematics; laser capture microdissection; male; microangiography; novel; public health relevance; sex; therapeutic angiogenesis; tibia

DESCRIPTION (provided by applicant): Massive bone and joint defects arise from resection of primary and metastatic tumors, congenital deficiency, traumatic loss, infection, or prosthetic implant failure. Available reconstructive methods are prone to high rates of failure. They include structural allografts (infection, nonunion and stress fracture), vascularized autografts (limited availability, size/shape mismatch, morbidity), or prosthetic replacement (infection, periprosthetic fracture and failure). A better solution to this difficult problem is required. Transplantation of living allogenic bone/joint would allow close matching of defect dimension and structure, while simultaneously maintaining the functional and healing properties of living tissue. Long-term immune modulation is necessary at present, unacceptable due to substantial health risks of immunosuppressive drugs or tolerance induction for such non- life critical transplantation. Maintaining allogeneic tissue viability without immunosuppression would be an important advance, and is the goal of this grant renewal proposal. Therapeutic angiogenesis is used to develop a host-derived neoangiogenic circulation within the transplanted bone/joint that maintains blood flow regardless of immune status. Previous work has demonstrated the method's promise. The Specific Aims of this proposal are #1) to test whether local delivery of vasculogenic growth factors improve measures of capillary formation and bone blood flow in living femoral allotransplants with simultaneous recipient AV bundle implantation, #2) to determine whether local delivery of a bone morphogenic protein may enhance new bone formation in the same model, #3 )to investigate whether osteocytes in newly formed bone are of recipient rather than transplant origin, and #4) to evaluate the viability, healing potential and function of orthotopic whole joint composite tissue transplants in a larger animal (rabbit knee) model. Improved clinical outcome of limb salvage surgery performed in difficult circumstances is the ultimate aim of this basic research. Methods: In aims 1, 2 and 3, sex-mismatched vascularized femora are transplanted using a saphenous arteriovenous (AV) bundle to provide host-derived angiogenesis. Growth factors are delivered, encapsulated within biodegradable microspheres placed within the transplanted femur. Angiogenesis is measured at two time points by quantifying capillary density with microangiography and bone blood flow by hydrogen washout; while bone remodeling is determined after fluorochrome labeling by quantitative histomorphometry. Laser capture microdissection will be used to study lineage of osteocytes specifically located in areas of newly formed bone, as measured by quantitative real-time PCR for the Y-chromosome-specific SRY gene (thus defining host or graft origin of the cells). Following whole knee joint transplants in a rabbit model, mechanical properties of joint cartilage and kinematic analysis of joint function will be correlated with measures of blood flow, angiogenesis and histology to assess the method's use in composite tissue allotransplantation. PUBLIC HEALTH RELEVANCE: Segmental bone defects are commonly encountered in orthopedic practice but available reconstructive methods, including structural allografts, vascularized autografts and prosthetic replacement are prone to high rates of failure. Transplantation of living allogenic bone and/or joint is a potential solution that would allow close matching of defect dimension and structure, while simultaneously maintaining the functional and healing properties of living tissue. The goal of this grant renewal proposal is to study the means, mechanism of action and effectiveness of a surgically-created host-derived neoangiogenic circulation to maintain blood flow and function in living musculoskeletal tissue transplants without need for prolonged postoperative drug therapy or tolerance induction.

描述（由申请人提供）：骨骼和关节缺陷由原发性和转移性肿瘤的切除，先天性缺乏，创伤性损失，感染或假体植入物衰竭引起。可用的重建方法容易出现高失败率。它们包括结构同种异体移植物（感染，骨不足和应力骨折），血管化自体移植（有限的可用性，尺寸/形状不匹配，发病率）或假体替代（感染，周围骨折和失败）。需要更好地解决这个困难问题。生命同种异体骨/关节的移植将允许与缺陷维度和结构密切匹配，同时维持活组织的功能和愈合特性。目前需要长期免疫调节，这是由于免疫抑制药物的重大健康风险或这种非生命关键移植的耐受性诱导而无法接受的。在没有免疫抑制的情况下保持同种异体组织的生存力将是一个重要的进步，这是该授予更新建议的目标。治疗性血管生成用于在移植的骨/关节内开发宿主衍生的新血管生成循环，无论免疫状态如何，它都保持血液流动。以前的工作已经证明了该方法的诺言。该提案的具体目的是＃1）测试血管生长生长因子的局部输送是否改善了与同时接收者同时植入的股骨同种异体移植物中毛细血管形成和骨血流动的度量，＃2）确定骨形蛋白在同一模型中是否可以增强骨形成的新骨形成，是否可以在同一模型中提高骨骼形式，是否会在同一模型中增强骨骼的新骨形成，是否会在同一模型中提高骨骼的效果。移植起源和＃4）评估较大动物（兔膝盖）模型中原位整体关节复合组织移植的生存力，愈合潜力和功能。这项基础研究的最终目的是在困难情况下进行的肢体救助手术的临床结果改善。方法：在目标1、2和3中，使用隐式动静脉（AV）束对性不匹配的血管化股骨进行移植，以提供宿主衍生的血管生成。生长因子被交付，封装在移植股骨内的可生物降解的微球内。血管生成是通过微观摄影和氢洗涤来量化毛细管密度和骨血流的两个时间点测量的；而通过定量组织形态法测定荧光体标记后骨重塑。激光捕获的微分解将用于研究特异性位于新形成骨区域的骨细胞的谱系，如Y染色体特异性SRY基因的定量实时PCR测量（从而定义了细胞的宿主或移植物来源）。在兔模型中，全膝关节移植后，关节软骨的机械性能和关节功能的运动学分析将与血流，血管生成和组织学的测量相关，以评估该方法在复合组织同种异体移植中的使用。 公共卫生相关性：骨科实践中通常会遇到分段骨缺陷，但可用的重建方法，包括结构性同种异体移植，血管化自体移植和假肢替代物，易于失败率很高。生命同种异体骨和/或关节的移植是一种潜在的解决方案，可以允许缺陷维度和结构紧密匹配，同时维持活性组织的功能和愈合特性。该赠款更新建议的目的是研究由手术衍生的宿主衍生的新血管生成循环的手段，作用机理和有效性，以维持血流和在活肌肉骨骼组织移植中的血液流量和功能，而无需延长术后术后治疗或耐受性诱导。