Tendon-Bone Construct Tissue Engineering for Extremity Reconstuction

用于四肢重建的肌腱-骨结构组织工程

• 批准号: 9114884
• 负责人: James Chang
• 金额: --
• 依托单位: VETERANS ADMIN PALO ALTO HEALTH CARE SYS
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9114884
• 关键词: Accidents; Allografting; Anatomy; Animal Model; Area; Athletic Injuries; Biocompatible; Biocompatible Materials; Biological Assay; Biological Preservation; Biomechanics; Biomedical Engineering; Blood Platelets; Body Temperature; Bone Regeneration; Bone Tissue; Bone Transplantation; Cadaver; Caring; Cell Proliferation; Cell Survival; Cells; Clinical; Complex; DNA; Defect; Degenerative Disorder; Development; Distal; Early Mobilizations; Extracellular Matrix Proteins; Fatty acid glycerol esters; Flexor; Gel; Goals; Grant; Healed; Histology; Human; Hydrogels; Immunohistochemistry; Implant; In Situ; In Situ Hybridization; In Vitro; Injury; Lead; Limb structure; Liquid substance; Measurement; Medical; Methods; Migration Assay; Military Personnel; Modeling; Molecular; Motion; Operating Rooms; Operative Surgical Procedures; Patients; Phalanx; Plasma; Postoperative Period; Powder dose form; Process; Production; Proliferating; Property; Protocols documentation; Rattus; Rehabilitation therapy; Research; Resected; Resolution; Roentgen Rays; Sampling; Seeds; Site; Soldier; Solid; Stem cells; Structure; Supplementation; Surgeon; Surgical sutures; Survival Rate; System; Techniques; Tendon structure; Testing; Time; Tissue Banking; Tissue Banks; Tissue Engineering; Tissues; Training; Translating; Translations; Upper Extremity; Veterans; achilles tendon; anterior cruciate ligament rupture; base; biomaterial compatibility; bone; bone healing; bone strength; calcaneum; cell motility; clinical application; combat; healing; human tissue; improved; improved outcome; in vivo; injured; joint function; microCT; migration; novel; public health relevance; reconstitution; reconstruction; repaired; rotator cuff tear; tendon rupture; tissue culture

 DESCRIPTION (provided by applicant): When soldiers and Veterans injure their extremities, tendons are likely to be disrupted. Without intact tendon-bone units, extremities cannot function properly. These injuries may be mid-substance tendon ruptures, or tears at the tendon-bone interface (TBI). The most difficult aspect of tendon reconstruction is adequate healing of this interface with bone. To date, the healing process of the TBI is poorly understood. The goals of this project are to determine the cellular and molecular mechanisms of TBI healing, and to develop tissue engineered TBI constructs that can be used for extremity reconstruction. It is hypothesized that tissue-engineered composite constructs consisting of tendon and bone would allow reconstruction with tendon-to-tendon and bone-to-bone healing. This would be faster and stronger, thus offering earlier mobilization and improved outcomes. Because there is a paucity of research on the process of TBI healing and tissue engineered TBI composite constructs, the following research objectives are proposed: The initial objective is to use an animal model of TBI healing based on the rat Achilles tendon to develop tendon-bone constructs. Decellularization of rat Achilles tendon-calcaneal bone segments and reseeding with adipoderived stem cells (ASCs) will be performed to create tissue engineered composite constructs. This process will be optimized with platelet-rich plasma (PRP) supplementation and the use of a novel tendon hydrogel as a carrier. The hydrogel is produced from decellularized tendon powder, is reconstituted into liquid form, and becomes a solid, yet porous gel at body temperature. It is hypothesized that the strength of bone-bone healing and tendon-tendon healing will exceed that of tendon-bone healing. Furthermore, repairs made with reseeded TBI grafts would be stronger than repairs performed with decellularized TBI grafts. In order to test this, tissue engineered rat Achilles TBI constructs will be re-implanted to assess fo short-term and long-term biocompatibility and repair strength. Techniques to assess healing will include immunohistochemistry, in situ hybridization, MTS measurement of tendon and bone-tendon junction biomechanical strength, high resolution Xray, and micro CT. Lastly, the ultimate goal would be translation to the production of human TBI composite grafts for clinical use in the reconstruction of extremity injuries. Human tissue engineered TBI constructs will be created based on optimized decellularization and reseeding techniques from the previous objectives. The specific anatomic structure to be produced initially is the distal phalanx-flexor tendon graft. The strength of native versus decellularized composite grafts will be tested. This proposal will ultimately allow the development of human composite TBI grafts for use in the reconstruction of traumatic and degenerative conditions of the extremities in soldiers and Veterans.

 描述（由申请人提供）： 当士兵和退伍军人伤害他们的四肢，肌腱很可能会中断。没有完整的腱骨单位，四肢不能正常工作。这些损伤可能是中间物质肌腱断裂，或肌腱-骨界面（TBI）撕裂。肌腱重建最困难的方面是充分愈合这个界面与骨。迄今为止，对TBI的愈合过程知之甚少。该项目的目标是确定TBI愈合的细胞和分子机制，并开发可用于肢体重建的组织工程TBI结构。 假设由肌腱和骨组成的组织工程复合结构将允许重建肌腱-肌腱和骨-骨愈合。这将更快、更有力，从而提供更早的动员和更好的结果。由于缺乏对TBI愈合过程和组织工程TBI复合结构的研究，提出了以下研究目标：最初的目标是使用基于大鼠跟腱的TBI愈合的动物模型来开发腱-骨结构。将大鼠跟腱-跟骨骨段脱细胞并用脂肪来源的干细胞（ASC）重新接种以产生组织工程复合结构。这一过程将通过补充富血小板血浆（PRP）和使用新型肌腱水凝胶作为载体来优化。该水凝胶由脱细胞肌腱粉制成，重组为液体形式，并在体温下成为固体但多孔的凝胶。 假设骨-骨愈合和腱-腱愈合的强度将超过腱-骨愈合。此外，用再灌注TBI移植物进行的修复将比用脱细胞TBI移植物进行的修复更强。为了测试这一点，组织工程大鼠跟腱TBI结构将被重新植入，以评估短期和长期的生物相容性和修复强度。评估愈合的技术将包括免疫组织化学、原位杂交、肌腱和骨-肌腱连接处生物力学强度的MTS测量、高分辨率X射线和微型CT。 最后，最终的目标将是转化为人类创伤性脑损伤复合移植物的生产，用于肢体损伤的重建。人类组织工程化TBI构建体将基于来自先前目标的优化的脱细胞化和再接种技术来创建。最初要产生的特定解剖结构是远节指骨-屈肌腱移植物。 将测试天然与脱细胞复合移植物的强度。 该提案最终将允许开发用于重建士兵和退伍军人四肢创伤和退行性疾病的人类复合TBI移植物。