A Novel Strategy of Endogenous Progenitor Cell Homing Using Exosomes for Condylar Fibrocartilage Repair in Temporomandibular Joint

利用外泌体进行内源性祖细胞归巢的新策略用于颞下颌关节髁纤维软骨修复

• 批准号: 10260595
• 负责人: Kyungsup Shin
• 金额: $ 15.45万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-10 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10260595
• 关键词: 3-Dimensional; Applications Grants; Arthralgia; Arthroscopy; Attention; Autologous; Bilateral; Biological Assay; Bone Marrow; Caring; Cell Communication; Cell Lineage; Cell Therapy; Cells; Characteristics; Chemotactic Factors; Chemotaxis; Chondrocytes; Chondrogenesis; Clinical; Clinical Treatment; Data; Degenerative Disorder; Degenerative polyarthritis; Diagnosis; Disease; Encapsulated; Fibrocartilages; Fossa; Functional disorder; Future; Goals; Histologic; Homing; Human; Hyaline Cartilage; Hyaluronic Acid; Hydrogels; In Situ; In Vitro; Injections; Injury; International; Intervention; Joints; Knowledge; Lead; Left; Lesion; Long-Term Effects; Mandible; Marshal; Masticatory muscles; MicroRNAs; Modeling; Morphology; Natural regeneration; Operative Surgical Procedures; Oral; Oryctolagus cuniculus; Outcome; Pain; Patients; Perforation; Play; Postoperative Period; Procedures; Production; Proliferating; Prunella vulgaris; Replacement Arthroplasty; Research; Role; Signs and Symptoms; Site; Societies; Stains; Stromal Cells; Structure of articular disc of temporomandibular joint; Surface; Symptoms; System; Temporomandibular Joint; Temporomandibular Joint Disorders; Temporomandibular Joint Dysfunction Syndrome; Testing; Therapeutic; Therapeutic Effect; Tissue Engineering; Tissues; Traumatic injury; United States; Work; arthropathies; base; cartilage regeneration; cartilage repair; cell motility; cost; design; disorder risk; engineered exosomes; exosome; healing; implantation; improved; in vivo; intercellular communication; joint injury; maxillofacial; miRNA expression profiling; minimally invasive; next generation sequencing; novel; novel strategies; prevent; protein biomarkers; recruit; regenerative; repair strategy; repaired; stem cells; tissue repair; treatment strategy; uptake

PROJECT SUMMARY/ABSTRACT Degenerative temporomandibular joint (TMJ) disorders (TMD) are some of the most challenging oral and maxillofacial problems, and when left untreated, they eventually lead to significant functional deficits, pain, stiffness, and osteoarthritis. As an example of degenerative TMD, the risk for post-traumatic osteoarthritis (PTOA) after serious joint injuries has not improved substantially in the last 40 years despite many refinements in care. This underscores the urgent need for new treatments to prevent PTOA initiated by joint damage. Fibrocartilage is notoriously limited in its intrinsic capacity for repair, focal damage associated with joint injuries seldom heals and often worsens to engulf entire articular surfaces. Cell-based therapies intended to regenerate neocartilage in situ have shown some clinical promise. The two most common, microfracture and autologous chondrocyte implantation (ACI), work well in the short term; however, long term results have been disappointing. The presence of chondrogenic progenitor cells (CPCs) in the TMJ fibrocartilage suggests that a rudimentary self- repair mechanism exists that might be marshaled for fibrocartilage regeneration after traumatic injury. CPC recruitment can be enhanced by exosomes derived from bone marrow stromal cells (BMSC-Exos), which serve an important role in intercellular communication and tissue repair. Our central hypothesis is that BMSC-Exos will stimulate TMJ condylar fibrocartilage repair by promoting CPC chemotaxis and chondrogenesis, intended for the treatment of focal lesions in TMJ fibrocartilage. The main appeal of a CPC-based repair strategy lies in their ability to heal by self-congregating at injury sites without ex vivo cell expansion and the additional surgery needed for grafting in ACI. Specific aims in this proposed study are to characterize BMSC-Exos and determine the effects of BMSC-Exos on CPC migration and fibrocartilage differentiation in vitro. Finally, in vivo fibrocartilage repair will be evaluated in a rabbit TMJ disc perforation model, where direct fibrocartilage contact between the condyle and mandibular fossa will induce degenerative fibrocartilage. BMSC-Exos encapsulated in hydrogel (F-127/hyaluronic acid) will be injected to the damaged TMJs 2 weeks after the perforation surgeries. Six weeks after the injection, integrity of the repaired fibrocartilage will be determined by histological/immunohistochemical staining and evaluated based on a modified OARSI scoring system. At the conclusion of this project, we will be able to define the therapeutic potential of BMSC- Exos for fibrocartilage repair and to identify miRNAs that may regulate cell migration and neotissue formation. As our future research plans, we will validate the candidate key miRNAs and synthesize target microRNA (miRNAs) for CPC migration and neofibrocartilage formation. The target miRNAs will be loaded in engineered exosomes for in vivo delivery, thereby replacing BMSC culture as a means of exosome production. This will further enhance the translatability of the minimally-invasive, single-step delivery procedure for cartilage regeneration described in this proposal.

项目概要/摘要 退行性颞下颌关节 (TMJ) 疾病 (TMD) 是最具挑战性的口腔和关节疾病之一。 颌面问题，如果不及时治疗，最终会导致严重的功能缺陷、疼痛、 僵硬和骨关节炎。作为退行性 TMD 的一个例子，创伤后骨关节炎 (PTOA) 的风险 尽管在护理方面进行了许多改进，但在过去 40 年中，严重关节损伤后的病情并没有得到实质性改善。 这强调了迫切需要新的治疗方法来预防关节损伤引起的 PTOA。纤维软骨 众所周知，其内在修复能力有限，与关节损伤相关的局灶性损伤很少愈合 并且经常恶化以吞没整个关节表面。旨在再生新软骨的细胞疗法 原位显示出一些临床前景。最常见的两种，微骨折和自体软骨细胞 植入（ACI），短期内效果良好；然而，长期结果却令人失望。这 颞下颌关节纤维软骨中软骨祖细胞（CPC）的存在表明，一个基本的自我 存在修复机制，可用于创伤性损伤后纤维软骨的再生。中国共产党 源自骨髓基质细胞的外泌体 (BMSC-Exos) 可以增强募集，其作用是 在细胞间通讯和组织修复中发挥重要作用。 我们的中心假设是 BMSC-Exos 将通过促进 TMJ 髁突纤维软骨修复来刺激 CPC 趋化性和软骨形成，用于治疗 TMJ 纤维软骨的局灶性病变。主要 基于 CPC 的修复策略的吸引力在于它们能够通过在损伤部位自我聚集来愈合，而无需前处理 ACI 中移植所需的体内细胞扩增和额外手术。这项拟议研究的具体目标 旨在表征 BMSC-Exos 并确定 BMSC-Exos 对 CPC 迁移和纤维软骨的影响 体外分化。最后，将在兔颞下颌关节盘穿孔模型中评估体内纤维软骨修复， 髁突和下颌窝之间的纤维软骨直接接触会导致退行性病变 纤维软骨。将封装在水凝胶（F-127/透明质酸）中的 BMSC-Exos 注射到受损部位 穿孔手术后 2 周的颞下颌关节。注射六周后，修复的纤维软骨变得完整 将通过组织学/免疫组织化学染色来确定，并根据修改后的 OARSI 进行评估 评分系统。在该项目结束时，我们将能够确定 BMSC 的治疗潜力 Exos 用于纤维软骨修复并鉴定可调节细胞迁移和新组织形成的 miRNA。 作为我们未来的研究计划，我们将验证候选关键miRNA并合成目标microRNA (miRNA) 用于 CPC 迁移和新纤维软骨形成。目标 miRNA 将被加载到工程中 外泌体用于体内递送，从而取代骨髓间充质干细胞培养作为外泌体生产的手段。这将 进一步增强微创、一步式软骨输送手术的可转化性 本提案中描述的再生。