Lim S A, Chen S, Suzuki I, Lambaren K, Soleimani A, Ho N, Mousavi M, Chung E J
Department of Biomedical Engineering, University of Southern California, Los Angeles, California 90089, United States.
Department of Medicine, Division of Nephrology and Hypertension, Keck School of Medicine, University of Southern California, Los Angeles, California 90033, United States.
ACS Biomater Sci Eng. 2025 Jul 14;11(7):4266-4278. doi: 10.1021/acsbiomaterials.5c00760. Epub 2025 Jun 27.
Natural killer cell-derived extracellular vesicles (NK-EVs) have demonstrated anti-inflammatory properties similar to those of their parent cells. EVs have been commonly delivered via intravenous (IV) administration, which can be invasive and is not ideal for chronic treatment. Another limitation of nanotherapy is its storage requirements, as EVs are commonly stored at -80 °C to preserve EV cargo and stability. In order to address these limitations, we explored dissolvable microneedles (MNs) as a promising alternative method for the administration of EVs. MNs have been used to deliver drugs, vaccines, and biomolecules, offering a convenient, noninvasive route of administration while preserving the therapeutic efficacy of EVs for extended periods, even at room temperature. Thus, we hypothesize that MN has the potential to sustain NK-EV stability and successfully deliver NK-EVs with minimal invasion. To test our hypothesis, we first developed an optimal MN formulation composed of hyaluronic acid and trehalose, both protein-protective materials that are biocompatible and biodegradable. After preparing MNs, we evaluated their stiffness, EV release profile, and ability to puncture pig skin. Additionally, the long-term storage stability of the EVs in MNs in inflammatory models in vitro and in vivo was evaluated. The MN successfully maintained EV efficacy even after storage after six months at room temperature, reducing the pro-inflammatory cytokine IL-6 by about 70% in inflamed human fibroblast cells relative to nontreated groups. Furthermore, EV-loaded MN treatment reduced both pro-inflammatory cytokines (IL-6 and TNFα) and psoriasis markers (Ki67 and IL-17) expression in a psoriasis model of chronic inflammation by about 40% compared to nontreated groups. Herein, our MN demonstrates the potential for easy-to-administer NK-EV therapies with long-term storage capabilities that preserve the NK-EV's anti-inflammatory properties.
自然杀伤细胞衍生的细胞外囊泡(NK-EVs)已表现出与其母细胞相似的抗炎特性。细胞外囊泡通常通过静脉内(IV)给药,这种给药方式具有侵入性,并不适合长期治疗。纳米疗法的另一个局限性在于其储存要求,因为细胞外囊泡通常储存在-80°C以保持其内容物和稳定性。为了解决这些局限性,我们探索了可溶解微针(MNs)作为一种有前景的细胞外囊泡给药替代方法。微针已被用于递送药物、疫苗和生物分子,提供了一种方便、非侵入性的给药途径,同时即使在室温下也能长时间保持细胞外囊泡的治疗效果。因此,我们假设微针有潜力维持自然杀伤细胞衍生的细胞外囊泡的稳定性,并以最小的侵入性成功递送自然杀伤细胞衍生的细胞外囊泡。为了验证我们的假设,我们首先开发了一种由透明质酸和海藻糖组成的最佳微针配方,这两种都是具有生物相容性和可生物降解性的蛋白质保护材料。制备微针后,我们评估了它们的硬度、细胞外囊泡释放曲线以及穿刺猪皮肤的能力。此外,还评估了细胞外囊泡在微针中在体外和体内炎症模型中的长期储存稳定性。即使在室温下储存六个月后,微针仍成功维持了细胞外囊泡的疗效;相对于未处理组,在发炎的人成纤维细胞中,微针使促炎细胞因子白细胞介素-6(IL-6)减少了约70%。此外,在慢性炎症的银屑病模型中,与未处理组相比,负载细胞外囊泡的微针治疗使促炎细胞因子(IL-6和肿瘤坏死因子α(TNFα))以及银屑病标志物(Ki67和IL-17)的表达均降低了约40%。在此,我们的微针展示了易于给药的自然杀伤细胞衍生的细胞外囊泡疗法的潜力,这种疗法具有长期储存能力,能够保留自然杀伤细胞衍生的细胞外囊泡的抗炎特性。
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