Dipartimento di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche (STEBICEF), Università degli Studi di Palermo, Parco d'Orleans II, Ed. 17, 90128 Palermo, Italy.
Dipartimento di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche (STEBICEF), Università degli Studi di Palermo, Parco d'Orleans II, Ed. 17, 90128 Palermo, Italy.
Colloids Surf B Biointerfaces. 2022 Dec;220:112931. doi: 10.1016/j.colsurfb.2022.112931. Epub 2022 Oct 14.
In the last years, the use of clay minerals for pharmaceutical purposes has increased due to their interesting properties. Hectorite (Ht) is a clay belonging to the smectite group which has attracted attention for applications in biology, tissue engineering and as drug carrier and delivery system. However, the mechanisms involved in Ht cellular uptake and transport into cells, are still unclear. Herein, we used a labeled Ht (Ht/1Cl) to study both the cellular uptake, by confocal laser scanning microscopy, and internalization pathways involved in the cellular uptake, by various endocytosis-inhibiting studies and fluorescence microscopy. These studies highlighted that Ht can penetrate the cellular membrane, localizing mainly in the cytoplasm. The main intracellular transport mechanisms are the ATP-dependent ones and those where filaments and microtubules are involved. Finally, as proof of concept for the potential of Ht as carrier system, we envisaged the covalent grafting of the anticancer molecule methotrexate (MTX), chosen as model, to obtain the Ht-MTX nanomaterial. The covalent linkage was confirmed by several techniques and the morphology of the obtained nanomaterial was imaged by SEM and TEM investigations. The kinetic release of the drug from the Ht-MTX nanomaterial in physiological conditions was studied as well. Furthermore, cytotoxic studies on different cell lines, namely, HL-60, HL-60R, MCF-7, 5637, UMUC3 and RT112 showed that Ht could be a promising material for anticancer therapy.
在过去的几年中,由于粘土矿物具有有趣的性质,其在制药方面的应用有所增加。海泡石(Ht)是属于蒙脱石组的粘土,由于其在生物学、组织工程以及作为药物载体和输送系统的应用而受到关注。然而,Ht 细胞摄取和进入细胞的转运机制仍不清楚。在此,我们使用标记的 Ht(Ht/1Cl)通过共聚焦激光扫描显微镜研究细胞摄取,并通过各种细胞内吞抑制研究和荧光显微镜研究参与细胞摄取的内化途径。这些研究强调了 Ht 可以穿透细胞膜,主要定位于细胞质中。主要的细胞内转运机制是依赖于 ATP 的机制和涉及丝和微管的机制。最后,作为 Ht 作为载体系统的潜在用途的概念验证,我们设想将抗癌分子甲氨蝶呤(MTX)共价接枝到 Ht 上,以获得 Ht-MTX 纳米材料。通过多种技术证实了共价键的连接,并通过 SEM 和 TEM 研究对获得的纳米材料的形态进行了成像。还研究了在生理条件下从 Ht-MTX 纳米材料中释放药物的动力学。此外,对不同细胞系,即 HL-60、HL-60R、MCF-7、5637、UMUC3 和 RT112 的细胞毒性研究表明,Ht 可能是一种有前途的抗癌治疗材料。