Gene and drug co-delivery is considered a promising strategy in combination cancer therapy. In this study, we fabricated Cu-Al layered double hydroxide (LDH) decorated with nanoscale ortho-, meta-, and para-poly(phenylenediamine), and afterward, doxorubicin (DOX) was loaded into nanoparticles. Finally, LDH was wrapped with plasmid CRISPR (pCRISPR). The modified nanohexagonals had particle sizes of approximately less than 100 nm with drug uptake up to 62.3%. pH-responsive nanocarriers exhibited higher cargo release in an acidic environment (pH 5.5) compared to a neutral buffer of pH 7.4. CLSM analysis demonstrated internalization in MEF-7 and HEK-293 cells with high transfection efficiency, further confirming them as promising nanostructures in cargo delivery. Besides, the potential usage of the cell-imprinted technique in order to increase the interactions to the cells, and enhance the cellular internalizations/transfections were assessed and showed a considerable increase (up to 72%) in the cellular internalizations. LDH nanostructures displayed high biocompatibility against HEK-293 and PC12 cells after 24 and 48 h. While they had no significant impact on the viability of MCF-7 cells with the assistance of the cell-imprinted method, these non-viral vectors significantly reduced the viability of HT-29 cells, showing their anti-cancer activity. Ortho, meta, and para forms of the poly(phenylenediamine)s decorated on the surface of the LDH showed transfection efficiencies up to 29.1, 28.2, and 40.1%, respectively. Therefore, this study could be considered the highest record of the pCRISPR delivery using a composite substrate in the literature.

基因和药物共同递送被认为是癌症联合治疗中一种很有前景的策略。在这项研究中，我们制造了 Cu-Al 层状双氢氧化物 (LDH)，并用纳米级的邻位、间位和对位修饰。聚苯二胺，然后将多柔比星 (DOX) 加载到纳米颗粒中。最后，用质粒 CRISPR (pCRISPR) 包裹 LDH。改性的纳米六方体的粒径大约小于 100 nm，药物吸收率高达 62.3%。与 pH 7.4 的中性缓冲液相比，pH 响应纳米载体在酸性环境（pH 5.5）中表现出更高的货物释放。CLSM 分析证明 MEF-7 和 HEK-293 细胞具有高转染效率的内化，进一步证实它们在货物递送中是有前途的纳米结构。此外，评估了细胞印迹技术在增加与细胞的相互作用和增强细胞内化/转染方面的潜在用途，并显示细胞内化显着增加（高达 72%）。LDH 纳米结构在 24 和 48 小时后显示出对 HEK-293 和 PC12 细胞的高生物相容性。虽然它们在细胞印迹方法的帮助下对 MCF-7 细胞的活力没有显着影响，但这些非病毒载体显着降低了 HT-29 细胞的活力，显示出它们的抗癌活性。修饰在 LDH 表面的聚苯二胺的邻位、间位和对位形式的转染效率分别高达 29.1%、28.2% 和 40.1%。因此，本研究可以被认为是文献中使用复合底物进行 pCRISPR 递送的最高记录。