Immunotargeting of reparative cells and theranostic nanosomes to cartilage lesions

在创伤性损伤或退行性关节炎中检测早期软骨损伤的能力一直有限， 在治疗可能更有益的情况下阻止治疗。蛋白多糖/糖蛋白的耗竭 在这些疾病中，软骨表面的破坏导致潜在的II型胶原(CII)的暴露。这 允许CII作为抗II型胶原的单抗的免疫识别靶点 (MabCII)。MabCII可以被荧光标记用于软骨损伤或软骨退变的诊断，或者可以 直接偶联到包裹药物的纳米体上，以局部传递到软骨损伤处。初步 有证据表明，可以使用类似的策略来靶向和招募修复性软骨细胞和间充质细胞 干细胞转移到受损部位。在这一应用中，MabCII将被用于综合治疗计划中 将修复细胞导向关节软骨和半月板软骨的病变。治疗效果观察 这些软骨的修复将通过一种创新的荧光关节镜进行监测。此外，我们还将 通过降低基质金属蛋白酶优化修复细胞在软骨损伤中的募集和整合 关节腔内注射核因子-kappaB的激活抑制物 Kb)途径被包裹在MabCII靶向纳米小体中。这些程序非常新颖，而且 关节损伤和疾病的诊断和治疗的范式转换。我们的目标是：(1)。诊断 以及关节表面和半月板软骨的损伤和退变区域的特征。 猪膝关节采用敏感、MabCII抗体引导的荧光关节镜(FA)方法。膝盖 家猪的关节大小、负重要求与人类关节非常相似， 软骨厚度，并将用于表征手术引起的半月板损伤和 关节软骨。损伤将通过使用荧光与荧光MabCII结合而可视化 关节镜检查是我们开发的一种新方法，并得到了组织病理学的证实。在FA之后 损伤的表征，(2)MabCII抗体将用于靶向传递和招募 修复细胞移植到膝关节软骨损伤区。我们将调查治疗方法 荧光MabCII靶向软骨细胞或骨髓间充质干细胞的实验研究 关节内注射脂肪组织到关节或半月板软骨的关节内 用FA监测手术损伤的细胞定位和存留情况。细胞之间的招募在 膝关节的软骨损伤将通过在膝关节表面的生物素/亲和素配体的创新系统来促进 修复细胞和与II型胶原结合的细胞的多价抗体募集。修复性组织将 通过组织病理学和RT-PCR进行基因表达分析。我们将(三)进一步优化 MabCII治疗软骨损伤后替代细胞的募集和整合 靶向纳米小体装载了一种抑制NF-κB途径的激活。产品的生产 基质金属蛋白酶(MMP)被认为是软骨基质降解的一个因素。逐渐减少 基质金属蛋白酶的生产将成为加强修复工作的典型目标。靶向MabCII的纳米小体 包裹一种选择性的人IKK-2抑制剂，一种核因子-κB途径的激活剂，将被用作局部 修复治疗前减少受损软骨损伤中MMPs产生的递送系统 细胞。好了！ 好了！ 好了！

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). MabCII can be fluorescently labeled for diagnosis of cartilage injury or cartilage degeneration or may be directly coupled to nanosomes encapsulating drugs for localized delivery to the cartilage lesion. Preliminary evidence shows a similar strategy can be used to target and recruit reparative chondrocytes and mesenchymal stem cells to the damaged site. In this application, MabCII will be used in a comprehensive treatment plan for directing reparative cells to lesions of the articular cartilage and meniscal cartilages. The therapeutic efficacy for repairing these cartilages will be monitored by an innovative fluorescent arthroscopy. In addition, we will optimize the recruitment and integration of the reparative cells in the cartilage lesions by reducing MMP production in the joint by intra-articular injection of an inhibitor of 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. Our aims are: (1). To diagnose and characterize 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). The knee of the domestic pig closely resembles a human joint in terms of joint size, weight-bearing requirements, and cartilage thickness and will be used in the characterization of surgically-induced injuries to meniscal and articular cartilages. The damage will be 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) MabCII antibody will be used to target delivery and recruitment of reparative cells to the damaged areas of the knee joint cartilages. We will investigate 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 will be 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 will be analyzed over time by histopathology and gene expression by RT-PCR. We will (3) further optimize 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. The production of matrix metalloproteinases (MMP) is known to be a 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 production of MMPs in damaged cartilage lesions prior to treatment with reparative cells. ! ! !