Lavín-López María Del Prado, Torres-Torresano Mónica, García-Cuesta Eva María, Soler-Palacios Blanca, Griera Mercedes, Martínez-Rovira Martín, Martínez-Rovira José Antonio, Rodríguez-Puyol Diego, de Frutos Sergio
Graphenano S.L., 30510 Yecla, Spain.
Department of Immunology and Oncology, National Center for Biotechnology, Consejo Superior de Investigaciones Científicas (CSIC), 28049 Madrid, Spain.
Nanomaterials (Basel). 2024 Dec 4;14(23):1945. doi: 10.3390/nano14231945.
We previously described GMC, a graphene-based nanomaterial obtained from carbon nanofibers (CNFs), to be biologically compatible and functional for therapeutic purposes. GMC can reduce triglycerides' content in vitro and in vivo and has other potential bio-functional effects on systemic cells and the potential utility to be used in living systems. Here, immunoreactivity was evaluated by adding GMC in suspension at the biologically functional concentrations, ranging from 10 to 60 µg/mL, for one or several days, to cultured lymphocytes (T, B, NK), either in basal or under stimulating conditions, and monocytes that were derived under culture conditions to pro-inflammatory (GM-MØ) or anti-inflammatory (M-MØ) macrophages. All stirpes were obtained from human peripheral mononuclear cells (PBMCs) from anonymized healthy donors. The viability (necrosis, apoptosis) and immunological activity of each progeny was analyzed using either flow cytometry and/or other analytical determinations. A concentration of 10 to 60 µg/mL GMC did not affect lymphocytes' viability, either in basal or active conditions, during one or more days of treatment. The viability and expression of the inflammatory interleukin IL-1β in the monocyte cell line THP-1 were not affected. Treatments with 10 or 20 µg/mL GMC on GM-MØ or M-MØ during or after their differentiation process promoted phagocytosis, but their viability and the release of the inflammatory marker activin A by GM-MØ were not affected. A concentration of 60 µg/mL GMC slightly increased macrophages' death and activity in some culture conditions. The present work demonstrates that GMC is safe or has minimal immunological activity when used in suspension at low concentrations for pre-clinical or clinical settings. Its biocompatibility will depend on the dose, formulation or way of administration and opens up the possibility to consider GMC or other CNF-based biomaterials for innovative therapeutic strategies.
我们之前描述过GMC,一种从碳纳米纤维(CNF)获得的基于石墨烯的纳米材料,具有生物相容性且具有治疗功能。GMC可在体外和体内降低甘油三酯含量,对全身细胞具有其他潜在的生物功能作用,并具有在生物系统中应用的潜在效用。在此,通过在生物功能浓度(10至60μg/mL)下将GMC悬浮液添加至培养的淋巴细胞(T细胞、B细胞、NK细胞)中1天或数天,无论是在基础条件下还是刺激条件下,以及将在培养条件下衍生的单核细胞分化为促炎性(GM-MØ)或抗炎性(M-MØ)巨噬细胞,来评估免疫反应性。所有品系均来自匿名健康供体的人外周血单核细胞(PBMC)。使用流式细胞术和/或其他分析测定法分析每个子代的活力(坏死、凋亡)和免疫活性。在治疗1天或多天期间,10至60μg/mL的GMC浓度在基础或活跃条件下均不影响淋巴细胞的活力。单核细胞系THP-1中炎性白细胞介素IL-1β的活力和表达不受影响。在GM-MØ或M-MØ分化过程中或之后,用10或20μg/mL GMC处理可促进吞噬作用,但它们的活力以及GM-MØ释放炎性标志物激活素A不受影响。在某些培养条件下,60μg/mL的GMC浓度会轻微增加巨噬细胞的死亡和活性。目前的研究表明,在临床前或临床环境中以低浓度悬浮使用时,GMC是安全的或具有最小的免疫活性。其生物相容性将取决于剂量、制剂或给药方式,并为考虑将GMC或其他基于CNF的生物材料用于创新治疗策略开辟了可能性。