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.

项目摘要/摘要 退化性颞下颌关节（TMJ）疾病（TMD）是一些最具挑战性的口腔， 上颌面问题，当未经处理时，它们最终导致功能不足，疼痛， 僵硬和骨关节炎。作为退化性TMD的一个例子，创伤后骨关节炎（PTOA）的风险 在过去的40年中，尽管有许多护理，但在过去40年中，严重的联合受伤并没有大幅改善。 这强调了迫切需要新治疗以防止因关节损害引发的PTOA。纤维球杆菌 众所周知，其内在的维修能力受到限制，与关节伤害相关的局灶性损害很少治愈 而且常常会使整个关节表面吞噬。基于细胞的疗法，旨在再生新能力 原位表现出了一些临床承诺。这两个最常见的是微裂缝和自体软骨细胞 植入（ACI），在短期内正常工作；但是，长期的结果令人失望。这 TMJ纤维状软骨中的软骨祖细胞（CPC）的存在表明，基本的自我 创伤性损伤后可能存在修复机制，以用于纤维球杆菌的再生。 CPC 可以通过从骨髓基质细胞（BMSC-exos）得出的外泌体来增强招聘 在细胞间通信和组织修复中的重要作用。 我们的中心假设是，BMSC-Exos将通过促进刺激TMJ Condolar纤维球修复 CPC趋化性和软骨生成，旨在治疗TMJ纤维球治疗局灶性病变。主 基于CPC的维修策略的吸引力在于它们通过在没有EX的伤害网站上进行自我控制来治愈的能力 体内细胞膨胀和ACI接枝所需的其他手术。在这项拟议的研究中的具体目的 要表征BMSC-EXO，并确定BMSC-Exos对CPC迁移和纤维球杆菌的影响 体外分化。最后，将在兔子TMJ盘穿孔模型中评估体内纤维球维修修复， con和下颌窝之间的直接纤维球接触将引起退行性 纤维球杆菌。封装在水凝胶（F-127/透明质酸）中的BMSC-EXO将被注入受损的 穿孔手术后2周，TMJS。注射后六周，修复的纤维球纤维的完整性 将由组织学/免疫组织化学染色确定，并根据改良的OARSI进行评估 评分系统。在该项目的结论中，我们将能够定义BMSC-的治疗潜力 用于纤维球修复的EXOS，并鉴定可能调节细胞迁移和新动物形成的miRNA。 作为未来的研究计划，我们将验证候选密钥miRNA并合成目标microRNA （miRNA）用于CPC迁移和新纤维球卫生的形成。目标miRNA将被加载到工程中 体内递送的外泌体，从而取代BMSC培养物作为外泌体产生的一种手段。这会 进一步增强软骨的最小侵入性单步递送程序的可翻译性 该提案中描述的再生。