An Innovative Two-Step Therapeutic Strategy to Maximize the Effect of Stem Cell Therapy for Post-Traumatic Osteoarthritis

创新的两步治疗策略可最大限度地发挥干细胞治疗创伤后骨关节炎的效果

• 批准号: 10643442
• 负责人: Hongsik Jake Cho
• 金额: --
• 依托单位: MEMPHIS VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10643442
• 关键词: Adipose tissue; Age; Aging; Animal Model; Anti-Inflammatory Agents; Antioxidants; Apoptosis; Area; Arthritis; Behavior; Binding; Biomechanics; Cartilage; Cell Count; Cell Death Induction; Cell Survival; Cell Therapy; Cell Transplantation; Cell surface; Cells; Cessation of life; Chondrocytes; Clinical; Coculture Techniques; Cytoprotection; Data; Degenerative polyarthritis; Disease; Dose; Encapsulated; Engineering; Environment; Etiology; Exhibits; Extracellular Matrix Degradation; Face; Fat-Soluble Vitamin; Future; Goals; Growth; Homing; Hydrophobic Interactions; Immunosuppression; In Vitro; Individual; Inflammation; Inflammation Mediators; Inflammatory; Injectable; Injections; Injury; Intervention; Joints; Knee; Knee Injuries; Knee Osteoarthritis; Knee joint; Lesion; Lifting; Lipids; Matrix Metalloproteinases; Mechanical Stress; Mechanics; Mediating; Mesenchymal Stem Cells; Methods; Modeling; Molecular; Mus; Natural regeneration; Occupational; Occupations; Operative Surgical Procedures; Oxidative Stress; Pain; Patient Recruitments; Patients; Pre-Clinical Model; Procedures; Property; Protocols documentation; Recovery; Regenerative Medicine; Reporting; Resistance; Risk; Route; Sex Differences; Sports; Stem cell transplant; Stress; System; Technology; Therapeutic; Therapeutic Agents; Therapeutic Effect; Time; Tissues; Training; Translational Research; Transplantation; Traumatic Arthropathy; Traumatic injury; Treatment Efficacy; Treatment Step; Veterans; Vitamin E; adipose derived stem cell; aged; bone; cartilage degradation; cell injury; chondroprotection; clinical application; combat; cytokine; design; disability; effective therapy; free radical oxygen; functional outcomes; healing; immunogenicity; implantation; improved; in vitro Model; in vivo; inflammatory milieu; inhibitor; innovation; joint inflammation; joint injury; joint stress; lipofection; mechanical load; military veteran; mouse model; nanoparticle; personalized approach; preconditioning; repaired; response; statistics; stem cell therapy; stem cells; subchondral bone; success; targeted delivery; transmission process; treatment strategy

PROJECT SUMMARY/ABSTRACT The etiology of osteoarthritis (OA) is multi-factorial. Abnormal and excessive cumulative joint stress results in post-traumatic osteoarthritis (PTOA). Approximately 30% of knee OA in Veterans is attributable to occupational activities, particularly jobs requiring kneeling or squatting in combination with heavy lifting, such as sport activity injuries. Currently, there is no effective therapy for OA patients. Recently, the stem cell therapy has shown promise to regenerate the damaged joint tissue. In our previous study, adipose-derived stem cells (ASCs) show great promise as therapeutic agents in regenerative medicine because of their multi-lineage potential, immunosuppressive activities, limited immunogenicity, and relative ease of growth in culture. However, there are several concerns that impede the clinical use of stem cell therapy in the inflammatory joint environment such as apoptosis, dosing, timing of intervention, homing efficacy, and route of delivery of ASCs. We have also found that NF-κB inhibitors such as TPCA-1 decrease inflammation in mechanically injured knee joints using our established PTOA mouse model and in vitro model. In this project, we aim our studies based on two points: 1) Pre-treatment of exogenously derived ASCs with antioxidant (such as Vitamin-E) before injection into the joint can lead to cyto-protective effects and resistance to apoptosis and toxic inflammatory factors after transplantation. 2) TPCA1-nanosome can improve the harsh condition in the arthritic knee joint by anti-inflammatory mechanism before transplantation of an exogenous stem cell. Therefore, we will show the synergistic effect of this two-step therapeutic application (anti- inflammatory nanosome treatment of the arthritic joint followed by transplantation of the Preconditioned ASCs). We have two aims: (1) Investigate the cytoprotective effects and therapeutic potential of preconditioned ASCs in a model of PTOA. In this aim, we will investigate the therapeutic effect of the antioxidant (Vit-E-Ns) pre-treated ASCs in vivo using our PTOA mouse model of knee overloading. In order to tailor this approach to the appropriate veteran population (active vs retired) we will use aged mice to investigate whether they exhibit different healing responses and mechanisms relative to younger mice. We will confirm the persistence of the ASCs in the joint, correlating implantation levels with the reduction and repair of damaged cartilage as well as optimizing the cell number and treatment interval. We will examine localization of the ASCs in the joint and assess joint inflammation and cartilage integrity. We will investigate treatment-associated changes in the biomechanical properties of subchondral bone and cartilage and its effects on pain-related behavior through functional analyses. We will also investigate the recovery mechanism using an ASC-chondrocyte co-culture system. (2) Demonstrate the synergistic therapeutic efficacy of the two-step application: TPCA1-Nanosome (TPCA1-Ns) injection prior to transplantation of the preconditioned ASCs in a mouse model of PTOA. In this aim, we will neutralize inflammation using TPCA1-Ns treatment followed by the utilization of Vit-E nanosome pretreated ASCs for transplantation into the PTOA mouse model established in our lab. In this Aim 3, we will examine the functional outcomes of the cartilage, bone, and synovial tissues after the two-step treatment consisting of the combined injection of the TPCA1-Ns and transplantation of the preconditioned ASCs. This two-step anti-inflammatory and antioxidant-nanosome treatment strategy can improve the toxic micro- environment of the arthritic knee joint via delivery of TPCA1-Ns, and also prolong the ASCs viability to obtain enough time to survive for their immune-suppressive activity. We believe that the data obtained from this project will serve as a basis for understanding the mechanism of PTOA and the therapeutic efficacy of transplanted ASCs. Also, this data will help contribute to developing strategies for clinical applications in the future.

项目摘要/摘要 骨关节炎（OA）的病因是多因素的。异常和过量的累积关节应力结果 在创伤后骨关节炎（PTOA）中。退伍军人中约有30％的膝盖OA归因于 从事活动，尤其是需要跪下或蹲在结合繁重的工作中的工作， 作为运动活动伤害。目前，OA患者尚无有效的疗法。最近，干细胞疗法 已经显示出有望再生受损的关节组织。在我们先前的研究中，脂肪衍生的干细胞 （ASC）由于其多部件而表现出作为再生医学治疗剂的巨大希望 潜力，免疫抑制活性，有限的免疫原性和培养物相对易于生长。 但是，有几个问题阻碍了炎症关节中干细胞疗法的临床使用 凋亡，剂量，干预时机，归巢效率和ASC交付途径等环境。 我们还发现，NF-κB抑制剂（例如TPCA-1）会减少机械损伤的膝盖的炎症 使用我们已建立的PTOA小鼠模型和体外模型的关节。 在这个项目中，我们基于两个点的研究目的：1）预处理外源衍生的ASC与 抗氧化剂（例如维生素-E）在注入关节之前会导致细胞保护作用和抗性 2）TPCA1纳米体可以改善刺激性 在移植外源之前，通过抗炎机制在artritic膝关节中的状况 干细胞。因此，我们将展示此两步治疗应用的协同作用（抗 炎症性纳米体治疗原性关节，然后移植预处理的ASC）。 我们有两个目标：（1）研究预处理的细胞保护作用和治疗潜力 ASC在PTOA模型中。在此目标中，我们将研究抗氧化剂（VIT-E-NS）的治疗作用 使用我们的PTOA鼠标膝关节超载的PTOA鼠标模型在体内进行预处理的ASC。为了量身定制这种方法 适当的退伍军人人口（主动与已退休），我们将使用老年小鼠调查它们是否暴露 相对于年轻小鼠的愈合反应和机制不同。我们将确认 关节中的ASC，将植入水平与减少和修复软骨的减少和修复相关 优化细胞数和治疗间隔。我们将检查ASC在关节中的定位，并 评估关节注入和软骨完整性。我们将研究与治疗相关的变化 软骨下骨和软骨的生物力学特性及其对疼痛相关行为的影响 功能分析。我们还将使用ASC-软骨细胞共培养研究恢复机制 系统。 （2）演示了两步应用的协同理论：TPCA1纳米体 （TPCA1-NS）在PTOA小鼠模型中移植预处理的ASC之前注射。在这个 目的，我们将使用TPCA1-NS处理，然后使用VIT-E纳米体进行中和注射 预处理ASC以移植到我们实验室中建立的PTOA小鼠模型中。在这个目标3中，我们将 两步处理后，检查软骨，骨骼和滑膜时机的功能结果 由TPCA1-NS的组合注射和预处理ASC的移植组成。这 两步的抗炎和抗氧化剂纳入策略可以改善有毒的微型 通过递送TPCA1-NS的Artritic膝关节环境，还延长了ASC的生存能力 足够的时间来生存其免疫抑制活性。 我们认为，从该项目获得的数据将是理解机制的基础 PTOA和移植ASC的治疗效率。此外，这些数据将有助于发展 将来的临床应用策略。