BLR&D Research Career Scientist Award Application

关节炎是这个国家导致残疾的主要原因。既往有关节损伤的患者有早期发病的风险 即使接受重建手术，也会发生创伤后骨关节炎(PTOA)。PTOA见于 比年龄相关的骨关节炎患者更年轻，在有较高发病率的人群中发病率较高 身体职业需求(即军事人员)。目前，PTOA治疗主要是针对疼痛，直到 关节衰竭和关节置换手术，这是一种与长期康复相关的昂贵治疗。这个 我研究的总体目标是开发一种靶向纳米药物，可以阻止软骨退化， 提高患者的生活质量，减少关节置换术的需求。检测早期软骨的能力 创伤性损伤或退行性关节炎的损害是有限的，在治疗可能的情况下阻止治疗 做更有益的事。这些疾病中软骨表面蛋白多糖/糖蛋白的耗竭 导致基本的II型胶原(CII)被揭开。这使得CII可以作为免疫学上的 II型胶原蛋白单抗(MabCII)的可识别靶标。荧光MabCII可用于 诊断软骨损伤或退变或MabCII偶联纳米体靶向药物 用于局部输送到软骨损伤处。我们的初步证据表明，类似的策略可以针对 将修复的软骨细胞或间充质干细胞招募到受损部位。因此，MabCII用于 引导修复细胞向关节软骨和半月板损伤的综合治疗方案 并通过一种创新的荧光关节镜进行监测。此外，招聘和整合 软骨损伤中修复细胞的数量通过减少基质金属蛋白酶(MMPs)而得到优化 关节内注射一种核活化的药物抑制物在关节内产生 核因子kappaB(NF-KB)途径包裹在MabCII靶向的纳米小体中。这些程序是 对于关节损伤和疾病的诊断和治疗，非常新颖和范式的转变。我在用 (1)关节面、半月板损伤和退行性变的早期诊断 用MabCII抗体引导的灵敏的荧光关节镜方法观察猪膝关节中的软骨 (FA)。我们的PTOA模型使用手术导致的膝关节半月板和关节软骨损伤。 家猪。猪膝关节在大小、承重要求和软骨结构上与人类关节极为相似。 厚度。损伤是通过使用荧光关节镜结合到荧光MabCII上而可视化的 我们开发的新程序，并得到了组织病理学的证实。在FA表征之后 损伤，(2)修复细胞通过MabCII靶向膝关节软骨损伤的区域 抗体。我们正在研究荧光、MabCII靶向软骨细胞或 关节腔内注射骨髓和脂肪来源的间充质干细胞 在关节或半月板软骨受到手术损伤的情况下，监测细胞定位和 英足总坚持。在膝关节软骨损伤处的细胞对细胞的募集是由一种创新的 修复细胞表面生物素/亲和素配基系统及多价抗体募集 结合II型胶原的细胞。随着时间的推移，对修复组织进行组织病理学和基因分析 逆转录-聚合酶链式反应表达。我们正在(3)进一步优化招聘和整合接班人 MabCII靶向纳米体载药抑制物治疗软骨损伤中的细胞 激活NF-κB通路以减少基质金属蛋白酶，基质金属蛋白酶是软骨降解的已知因素 矩阵。减少基质金属蛋白酶的产生将是加强修复努力的典型目标。MabCII- 靶向纳米小体包裹了人IKK-2的选择性抑制物，人IKK-2是NF-κB途径的激活剂， 将被用作局部递送系统，在接受治疗之前减少受损软骨病变中的MMPs 修复细胞增强其在退变部位的存活率。 好了！

Arthritis is the nation's leading cause of disability. Patients with a previous joint injury are at risk for early development of post-traumatic osteoarthritis (PTOA) even with reconstructive surgery. PTOA is seen in younger patients than age-related osteoarthritis and it has a higher incidence in populations that have high physical occupational demands (i.e., military personnel). Currently, PTOA treatment is primarily for pain until joints fail and joint replacement surgery, an expensive treatment associated with a long rehabilitation. The overall goal of my research is to develop a targeted nanomedicine that can halt cartilage degeneration, improve quality of life and reduce the need for joint replacement in patients. The ability to detect early cartilage damage in traumatic injury or degenerative arthritis has been limited, preventing treatment when therapies may be more beneficial. Depletion of proteoglycans/glycoproteins on the surface of the cartilage in these disorders results in unmasking of the underlying type II collagen (CII). This allows CII to serve as an immunologically recognizable target for monoclonal antibody to type II collagen (MabCII). Fluorescent MabCII can be used for diagnosis of cartilage injury or degeneration or MabCII coupled to nanosomes can target encapsulated drugs for localized delivery to the cartilage lesion. Our preliminary evidence shows a similar strategy can target and recruit reparative chondrocytes or mesenchymal stem cells to the damaged site. Thus, MabCII is used in a comprehensive treatment plan for directing reparative cells to lesions of the articular cartilage and meniscal cartilages and monitoring this by an innovative fluorescent arthroscopy. In addition, recruitment and integration of the reparative cells in the cartilage lesions is optimized by reducing matrix metalloproteinases (MMP) production in the joint by intra-articular injection of a pharmacological inhibitor of the activation of the nuclear factor kappa B (NF-KB) pathway encapsulated in MabCII-targeted nanosomes. These procedures are extremely novel and paradigm shifting for the diagnosis and treatment of joint injury and disease. I am using them for: (1) early diagnosis of damaged and degenerative areas of articular surface and meniscal cartilages in the pig knee using a sensitive, MabCII antibody-guided method of fluorescent arthroscopy (FA). Our PTOA model uses surgically-induced injuries to meniscal and articular cartilages in the knee of the domestic pig. The pig knee closely resembles a human joint in size, weight-bearing requirements and cartilage thickness. The damage is visualized through its binding to fluorescent MabCII using fluorescent arthroscopy, a new procedure that we have developed, and confirmed by histopathology. After FA characterization of the injury, (2) reparative cells are targeted to the area of damaged cartilages in the knee with MabCII antibody. We are investigating the therapeutic efficacy of fluorescent, MabCII-targeted chondrocytes or mesenchymal stem cells derived from bone marrow and adipose tissues intra-articularly injected into joints where articular or meniscal cartilages have been surgically damaged monitoring the cellular localization and persistence by FA. Cell to cell recruitment at the cartilage lesion in the knee is facilitated by an innovative system of biotin/avidin ligands on the surface of the reparative cells and multivalent antibody recruitment of cells binding type II collagen. Reparative tissues are analyzed over time by histopathology and gene expression by RT-PCR. We are (3) further optimizing the recruitment and integration of replacement cells in the cartilage lesion by treatment with MabCII-targeted nanosomes loaded with an inhibitor of the activation of the NF-κB pathway to minimize MMP, a known factor in degradation of cartilage matrices. Diminishing MMP production will be a prototypic target for enhancing reparative efforts. The MabCII- targeted nanosomes encapsulating a selective inhibitor of human IKK-2, an activator of the NF-κB pathway, will be used as a local delivery system to reduce MMPs in damaged cartilage lesions prior to treatment with reparative cells enhancing their survival at the degenerative site. !