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.

项目总结/摘要 骨关节炎（OA）的病因是多因素的。异常和过度的累积关节应力结果 创伤后骨关节炎（PTOA）。退伍军人中约30%的膝关节OA可归因于 职业活动，特别是需要跪着或蹲着并同时举起重物的工作， 运动伤害。目前，对于OA患者没有有效的治疗方法。最近，干细胞疗法 有望使受损的关节组织再生在我们之前的研究中， 由于其多谱系性，ASCs作为再生医学中的治疗剂显示出巨大的前景 潜在的免疫抑制活性、有限的免疫原性和在培养物中相对容易生长。 然而，有几个问题阻碍了干细胞疗法在炎症关节中的临床应用 环境如细胞凋亡、剂量、干预时机、归巢功效和ASC的递送途径。 我们还发现NF-κB抑制剂如TPCA-1可减少机械损伤膝关节的炎症 关节使用我们建立的PTOA小鼠模型和体外模型。 在本项目中，我们的研究基于两点：1）用 抗氧化剂（如维生素E）注射到关节前，可导致细胞保护作用和抵抗力 移植后细胞凋亡和毒性炎症因子。2)TPCA 1-nanosome可以改善粗糙的 在移植外源性的抗炎症机制之前， 干细胞因此，我们将展示这种两步治疗应用的协同效应（抗- 炎性纳米体治疗关节炎关节，随后移植预处理的ASC）。 本研究的目的有两个：（1）探讨预处理的细胞保护作用和治疗潜力 PTOA模型中的ASC。为此，我们将研究抗氧化剂（Vit-E-Ns）的治疗作用 使用我们的膝关节超负荷的PTOA小鼠模型在体内预处理的ASC。为了使这种方法适合于 适当的退伍军人群体（活跃与退休），我们将使用老年小鼠来调查他们是否表现出 不同的愈合反应和机制相对于年轻的小鼠。我们将确认 关节中的ASC，将植入水平与受损软骨的减少和修复以及 优化细胞数量和处理间隔。我们将研究关节中ASCs的定位， 评估关节炎症和软骨完整性。我们将研究治疗相关的变化， 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-软骨细胞共培养来研究恢复机制。 系统(2)证明两步应用的协同治疗功效：TPCA 1-纳米体 在PTOA小鼠模型中移植预处理的ASC之前，注射TPCA 1-Ns。在这 目的是，我们将使用TPCA 1-Ns治疗中和炎症，然后使用Vit-E纳米体 预处理的ASC用于移植到我们实验室建立的PTOA小鼠模型中。在本目标3中，我们将 检查两步治疗后软骨、骨和滑膜组织的功能结果 包括联合注射TPCA 1-Ns和移植预处理的ASCs。这 两步抗炎和抗氧化纳米体治疗策略可以改善毒性微 通过递送TPCA 1-N，可以改善关节炎膝关节的环境，并且还延长ASC的存活力，以获得 有足够的时间来维持免疫抑制活性。 我们相信，从这个项目中获得的数据将作为了解 PTOA和移植ASCs的治疗效果。此外，这些数据将有助于开发 未来的临床应用策略。