Tekade Muktika, Sharma Mukesh Chandra
School of Pharmacy, Devi Ahilya Vishwavidyalaya, Takshila Campus, Khandwa Road, Indore, Madhya Pradesh India.
Indian J Microbiol. 2025 Mar;65(1):477-504. doi: 10.1007/s12088-024-01338-y. Epub 2024 Jul 29.
Rheumatoid arthritis (RA) is an autoimmune disorder that induces joint inflammation, cartilage injury, and bone damage. Thus far, methotrexate (MTX) is a primary DMARD drug to treat RA. Despite high efficiency, its clinical application is compromised due to delivery-associated systemic side effects. This investigation reports a Quality-by-design (QbD; Box Behnken Strategy) assisted production of a novel, innovative, and multipurpose polycation-templated approach for producing stable albumin MTX nanoparticles (T-AMTX-NP). This approach formed a highly biocompatible MTX formulation with reduced toxicity (1.81 ± 0.54% hemolysis) compared to plain MTX (13.19 ± 2.77%; SEM:1.965). T-AMTX-NP was found to be nanometric (Particle size: 135.86 ± 5.17 nm; PDI: 0.27) with a net surface charge of ζ -10.15 ± 2.19 mV. With 4.01-fold cationization (TNBS assay), T-AMTX-NP showed high drug loading (64.98 ± 1.25%) and sustained MTX release under physiological conditions (up to 48 h; < 0.001). The nanoformulation followed the Higuchi release kinetics model (R, 0.9957). Significantly reduced burst release by 70.61% ( = 0.0002) and 12.89% ( = 0.0115) compared to plain MTX and conventional MTX-formulation (AMTX-NP), respectively. Interestingly, T-AMTX-NP showed pH-responsive drug release bio-environment-responsive architectural change. Cell line studies in lipopolysaccharide (LPS) stimulated RAW264.7 macrophage showed a significant reduction in intracellular nitrite level following T-AMTX-NP treatment ( < 0.01). Cellular uptake and cell viability confirmed selective cellular uptake potential in inflamed cells. Furthermore, compared to the control, the high intracellular ROS-generation was noted with T-AMTX-NP (2.1485-fold; < 0.01). Furthermore, hemolysis assay and stability assessments were also conducted to determine the hemocompatibility and suitable conditions for the storage of nanoformulation. The outcome of this study suggests that the developed multipurpose nanoformulation is a superior therapeutics approach for improved RA treatment. Suggestively, the developed strategy can also be adopted to benefit other clinical situations that demand to counter inflammation, cytostatic as well as psoriatic conditions.
类风湿性关节炎（RA）是一种自身免疫性疾病，可引发关节炎症、软骨损伤和骨质破坏。到目前为止，甲氨蝶呤（MTX）是治疗RA的主要抗风湿药物。尽管其疗效高，但其临床应用因与给药相关的全身副作用而受到影响。本研究报告了一种基于质量源于设计（QbD；Box-Behnken策略）辅助生产的新型、创新且多功能的聚阳离子模板法，用于制备稳定的白蛋白MTX纳米颗粒（T-AMTX-NP）。与普通MTX（13.19±2.77%；SEM：1.965）相比，该方法形成了一种具有高生物相容性且毒性降低（溶血率1.81±0.54%）的MTX制剂。发现T-AMTX-NP为纳米级（粒径：135.86±5.17nm；PDI：0.27），净表面电荷为ζ-10.15±2.19mV。经4.01倍阳离子化（TNBS测定）后，T-AMTX-NP显示出高载药量（64.98±1.25%），并在生理条件下持续释放MTX（长达48小时；<0.001）。该纳米制剂遵循Higuchi释放动力学模型（R，0.9957）。与普通MTX和传统MTX制剂（AMTX-NP）相比，突释分别显著降低了70.61%（=0.0002）和12.89%（=0.0115）。有趣的是，T-AMTX-NP显示出pH响应性药物释放和生物环境响应性结构变化。在脂多糖（LPS）刺激的RAW264.7巨噬细胞系研究中，T-AMTX-NP处理后细胞内亚硝酸盐水平显著降低（<0.01）。细胞摄取和细胞活力证实了其在炎症细胞中的选择性细胞摄取潜力。此外，与对照组相比，T-AMTX-NP的细胞内ROS生成量较高（2.1485倍；<0.01）。此外，还进行了溶血试验和稳定性评估，以确定纳米制剂的血液相容性和合适的储存条件。本研究结果表明，所开发的多功能纳米制剂是一种用于改善RA治疗的卓越治疗方法。提示所开发的策略也可用于其他需要对抗炎症、细胞抑制以及银屑病状况的临床情况。
