Moghadam Naghmeh Akbari, Bagheri Fatemeh, Eslaminejad Mohamadreza Baghaban
Department of Biomedical Engineering, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, Iran.
Department of Biotechnology, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, Iran.
Colloids Surf B Biointerfaces. 2022 Nov;219:112786. doi: 10.1016/j.colsurfb.2022.112786. Epub 2022 Aug 23.
Conventional treatments for osteoarthritis (OA), including drug delivery and tissue engineering approaches, could not offer a high yield of cartilage repair due to the compact and exclusive structure of cartilage. Targeted and high-efficiency delivery of gene sequences is necessary to rebalance the lost homeostatic properties of the cartilage in OA. Herein, we synthesized chitosan (CH)-chondroitin sulfate (CS) nanoparticles (NPs) as a platform for delivering gene sequences. These new nanoparticles benefit from two natural polymers that minimize the toxicity, and the presence of CS can be in favor of targeted delivery. The CAG-GFP plasmid was used as a gene sequence model, and the nanoparticles could successfully encapsulate approximately all of them in their structure. Loaded nanoparticles were characterized in terms of morphology, size, zeta potential, the efficiency of encapsulation and, DNA release pattern. Cell viability and uptake of new nanoparticles were compared to the chitosan nanoparticles and Lipofectamine. After substituting TPP with CS, NPs exhibited a significant decrease in size. In addition, there was little difference in zeta potential between nanoparticles. Furthermore, a tremendous increase in plasmid uptake and cell viability was observed by CH-CS NPs compared to CH-TPP NPs and Lipofectamine. In the final stage, the knockdown level of MMP13 was evaluated with real-time RT-PCR for confirming the potential uptake of CH-CS NPs. The results revealed cellular uptake of siRNA loaded NPs and effective knockdown of MMP13 in chondrocytes. In conclusion, CH-CS nanoparticles can be considered as a candidate for gene therapy purposes in cartilage diseases.
骨关节炎（OA）的传统治疗方法，包括药物递送和组织工程方法，由于软骨致密且独特的结构，无法实现高产量的软骨修复。靶向和高效递送基因序列对于重新平衡骨关节炎中软骨丧失的稳态特性是必要的。在此，我们合成了壳聚糖（CH）-硫酸软骨素（CS）纳米颗粒（NPs）作为递送基因序列的平台。这些新型纳米颗粒受益于两种天然聚合物，可将毒性降至最低，并且CS的存在有利于靶向递送。CAG-GFP质粒用作基因序列模型，纳米颗粒能够成功地将几乎所有质粒包裹在其结构中。对负载的纳米颗粒进行了形态、大小、zeta电位、包封效率和DNA释放模式等方面的表征。将新型纳米颗粒的细胞活力和摄取情况与壳聚糖纳米颗粒和脂质体转染试剂进行了比较。用CS替代TPP后，NPs的尺寸显著减小。此外，纳米颗粒之间的zeta电位差异不大。此外，与CH-TPP NPs和脂质体转染试剂相比，CH-CS NPs观察到质粒摄取和细胞活力有极大提高。在最后阶段，通过实时RT-PCR评估MMP13的敲低水平，以确认CH-CS NPs的潜在摄取情况。结果显示负载siRNA的NPs被细胞摄取，并在软骨细胞中有效敲低了MMP13。总之，CH-CS纳米颗粒可被视为软骨疾病基因治疗的候选物。
