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.

项目总结/摘要 退行性颞下颌关节（TMJ）疾病（TMD）是一些最具挑战性的口腔和 颌面问题，如果不治疗，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） 在过去的40年里，尽管在护理方面进行了许多改进，但严重关节损伤后的关节炎并没有得到实质性的改善。 这强调了迫切需要新的治疗方法来预防由关节损伤引起的PTOA。纤维软骨 众所周知，关节的内在修复能力有限，与关节损伤相关的局灶性损伤很少愈合 并且经常会吞噬整个关节面。旨在再生新软骨的基于细胞的疗法 已经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）在短期内效果良好;然而，长期结果令人失望。的 TMJ纤维软骨中存在软骨形成祖细胞（CPC）表明， 创伤性损伤后纤维软骨再生可能存在修复机制。中共 来源于骨髓基质细胞的外来体（BMSC-Exos）可以增强募集， 在细胞间通讯和组织修复中起重要作用。 我们的中心假设是BMSC-Exos将通过促进TMJ髁状突纤维软骨修复来刺激TMJ髁状突纤维软骨修复。 CPC趋化性和软骨形成，用于治疗TMJ纤维软骨中的局灶性病变。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而愈合， 体内细胞扩增和ACI中移植所需的额外手术。这项拟议研究的具体目标 是表征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TMJ椎间盘穿孔模型中评价体内纤维软骨修复， 髁突和下颌窝之间的直接纤维软骨接触将诱发退行性变 纤维软骨包封在水凝胶（F-127/透明质酸）中的BMSC-Exos将被注射到受损的 颞下颌关节穿孔手术后2周。注射后6周，修复的纤维软骨的完整性 将通过组织学/免疫组织化学染色确定，并基于改良的OARSI进行评价 评分系统在该项目结束时，我们将能够确定BMSC的治疗潜力- 用于纤维软骨修复的Exos，并鉴定可能调节细胞迁移和新组织形成的miRNAs。 作为我们未来的研究计划，我们将验证候选的关键miRNAs并合成目标microRNA （miRNAs）用于CPC迁移和新纤维软骨形成。靶miRNA将被装载在工程化的细胞中， 在一些实施方案中，BMSC培养物可用于体内递送外泌体，从而取代BMSC培养物作为外泌体产生的手段。这将 进一步增强了软骨的微创、单步递送过程的可平移性 在这篇文章中描述的再生。