离子导入增强的粘膜腔内递送顺铂包封壳聚糖纳米粒用于鼠模型口腔癌治疗

顺铂是口腔癌治疗中最有效的化疗药物之一，但全身给药具有副作用。本研究旨在评估离子导入对增强顺铂从顺铂包封壳聚糖纳米粒中释放的影响。研究了不同质量比的壳聚糖与三聚磷酸盐（TPP）（5:1、10:1、15:1、20:1）对顺铂包封效率的影响。采用共聚焦激光扫描显微镜观察顺铂包封壳聚糖被细胞的摄取情况。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑盐（MTT）法检测不同顺铂浓度下的细胞存活率。采用恒流电流计（CCCP）、循环恒流计（CCP）和差分脉冲伏安法（DPV）三种离子导入方法，以增强顺铂从包封壳聚糖纳米粒中的释放。此外，将鼠口腔鳞状细胞癌细胞系植入鼠口腔黏膜以诱导口腔癌。评估CCCP、CCP和DPV增强顺铂释放对小鼠肿瘤抑制的效果。牺牲后，分离肿瘤和淋巴结进行苏木精-伊红染色和免疫组化染色，包括Ki-67和pan CK。采用电感耦合等离子体质谱法定量肿瘤中的铂含量。结果显示，质量比为15:1的纳米粒子在磷酸盐缓冲液中具有最高的顺铂包封效率（约15.6%）和最长的持续释放时间（最长达35天），释放率达100%。细胞摄取结果表明，壳聚糖纳米粒子通过内吞作用被运送到细胞质中。MTT试验结果显示，随着顺铂浓度的增加，细胞存活率下降。CCP（1 mA，开：关=1 s:1 s）和DPV（0-0.06 V）组在抑制肿瘤生长方面效果最佳，两组的Ki-67阳性和pan CK阳性比例最低。本研究首次探讨并验证了DPV在增强纳米粒子体内药物释放以治疗动物癌症中的潜力。结果提示，CCP和DPV方法有望与手术联合应用于口腔癌治疗。

Cisplatin is one of the most effective chemotherapeutic drugs used in oral cancer treatment, but systemic administration has side effects. The purpose of this study was to evaluate the effect of iontophoresis on the enhancement of cisplatin release from cisplatin-encapsulated chitosan nanoparticles.The effect of different mass ratios of chitosan to tripolyphosphate (TPP) (5:1, 10:1, 15:1, 20:1) on the encapsulation efficiency of cisplatin was investigated. Uptake of cisplatin-encapsulated chitosan by cells was observed using a confocal laser scanning microscope. The cell viability at different cisplatin concentrations was examined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Three iontophoresis methods, namely constant-current chronopotentiometry (CCCP), cyclic chronopotentiometry (CCP), and differential pulse voltammetry (DPV), were used to enhance cisplatin release from cisplatin-encapsulated chitosan nanoparticles. In addition, mouse oral squamous cell carcinoma cell lines were implanted into the mouse oral mucosa to induce oral cancer. The effects of enhanced cisplatin release by CCCP, CCP, and DPV on tumor suppression in mice were evaluated. Tumors and lymph nodes were isolated for hematoxylin-eosin staining and immunohistochemistry staining including Ki-67 and pan CK after sacrifice. Inductively coupled plasma mass spectrometry was conducted to quantify the platinum content within the tumors.The results showed that nanoparticles with a mass ratio of 15:1 exhibited the highest cisplatin encapsulation efficiency (approximately 15.6%) and longest continued release (up to 35 days) in phosphate buffered saline with a release rate of 100%. Cellular uptake results suggested that chitosan nanoparticles were delivered to the cytoplasm via endocytosis. The results of the MTT assay revealed that the survival rate of cells decreased as the cisplatin concentration increased. The CCP (1 mA, on:off = 1 s: 1 s) and DPV (0-0.06 V) groups were the most effective in inhibiting tumor growth, and both groups exhibited the lowest percentage of Ki-67 positive and pan CK positive.This study is the first to investigate and determine the efficacy of DPV in enhancing in vivo drug release from nanoparticles for the treatment of cancer in animals. The results suggest that the CCP and DPV methods have the potential to be combined with surgery for oral cancer treatment.