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: 9502188
• 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/9502188
• 关键词: Accidents; Allografting; Anatomy; Animal Model; Area; Athletic Injuries; Biocompatible Materials; Biological Assay; 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; Histologic; 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 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; experimental group; healing; human tissue; improved; improved outcome; in vivo; injured; joint function; microCT; migration; novel; preservation; 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）： 当士兵和退伍军人四肢受伤时，肌腱可能会受到破坏。如果没有完整的腱骨单位，四肢就无法正常运作。这些损伤可能是肌腱中部断裂或肌腱-骨界面撕裂 (TBI)。肌腱重建最困难的方面是与骨的界面的充分愈合。迄今为止，人们对 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，并开发可用于四肢重建的组织工程 TBI 结构。 据推测，由肌腱和骨骼组成的组织工程复合结构将允许通过肌腱到肌腱和骨骼到骨骼的愈合进行重建。这将更快、更强，从而提供更早的动员和更好的结果。由于对 TBI 愈合过程和组织工程 TBI 复合结构的研究很少，因此提出以下研究目标： 初步目标是使用基于大鼠跟腱的 TBI 愈合动物模型来开发腱骨结构。将对大鼠跟腱-跟骨骨段进行脱细胞化并重新接种脂肪干细胞（ASC），以创建组织工程复合结构。该过程将通过补充富含血小板的血浆（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末制成，被重组为液体形式，并在体温下变成固体但多孔的凝胶。 据推测，骨-骨愈合和腱-腱愈合的强度将超过腱-骨愈合。此外，用重新接种的 TBI 移植物进行的修复比用脱细胞 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，以评估短期和长期生物相容性和修复强度。评估愈合的技术包括免疫组织化学、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 射线和显微 CT。 最后，最终目标是转化为人类 TBI 复合移植物的生产，用于临床四肢损伤的重建。人体组织工程 TBI 构建体将基于先前目标的优化脱细胞和重新接种技术来创建。最初要制作的具体解剖结构是远端指骨屈肌腱移植物。 将测试天然复合移植物与脱细胞复合移植物的强度。 该提案最终将允许开发人类复合 TBI 移植物，用于重建士兵和退伍军人的四肢创伤和退行性疾病。

项目成果

Analysis of Cell-seeded, Collagen-rich Hydrogel for Wound Healing.

用于伤口愈合的细胞接种、富含胶原蛋白的水凝胶的分析。

• DOI: 10.1097/gox.0000000000003049
• 发表时间: 2020
• 期刊: Plastic and reconstructive surgery. Global open
• 作者: SoteloLeon,Daniel;Williams,Tokoya;Wang,Zhen;Leyden,Jacinta;Franklin,Austin;Kaizawa,Yukitoshi;Chang,James;Fox,PaigeM
• 通讯作者: Fox,PaigeM

Optimized Repopulation of Tendon Hydrogel: Synergistic Effects of Growth Factor Combinations and Adipose-Derived Stem Cells.

• DOI: 10.1177/1558944715628005
• 发表时间: 2017-01-01
• 期刊: Hand (New York, N.Y.)
• 作者: Farnebo, Simon;Farnebo, Lovisa;Chang, James
• 通讯作者: Chang, James