Shohani Sepideh, Mondanizadeh Mahdieh, Abdoli Asghar, Khansarinejad Behzad, Salimi-Asl Mohammad, Ardestani Mehdi Shafiee, Ghanbari Maryam, Haj Mehrdad Sadeghi, Zabihollahi Rezvan
Department of Biotechnology and Molecular Medicine, Faculty of Medicine, Arak University of Medical Science, P.O. Box: 6941, Arak, Iran.
Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran.
Curr HIV Res. 2017;15(1):56-65. doi: 10.2174/1570162X14666161216142806.
Highly active antiretroviral therapy (HAART) has been commonly used for HIV treatment. Its main drawbacks like drug resistance and side effects raised researcher's interest to find new approaches for its treatment. Trimethyl chitosan is one of the drug carriers which has been introduced recently.
the conjugated atripla-trimethyl chitosan was designed and characterized by zetasizer, AFM and FTIR techniques. The drug conjugation with trimethyl chitosan and cellular uptake of nano-conjugate were determined by spectrophotometry. XTT test was used to measure the cytotoxicity. Anti-retroviral efficiency was studied by ELISA test.
Zetasizer Results proved that the average size of nano-conjugate particles agglomeration was 493.4±24.6 nm but the size of the majority of the particles was 177.2±7.8 nm with the intensity of 87.9%. AFM technique revealed that the sizes of nano-conjugate and trimethyl chitosan were 129 nm and 59.78 nm, respectively. Zeta potential was -1.35±0.04 mv for nano-conjugate and -7.69±0.3 mv for drug. Conjugation efficiency of atripla with trimethyl chitosan was 5.27%. Measured cellular uptake with spectrophotometry for nano-conjugate was about twice of the free drug in examined concentrations (P=0.007). Compared to atripla, the nano-conjugate showed a higher inhibitory effect on HIV replication (P=0.0001).
The result showed that atripla-TMC conjugate does not have a significant cytotoxicity effect. Due to the higher inhibitory effect of nano-conjugate on viral replication, it can be used in lower concentration for antiviral treatment, which resulted in reduction of drug resistance and other side effects.
高效抗逆转录病毒疗法（HAART）已普遍用于治疗HIV。其耐药性和副作用等主要缺点激发了研究人员寻找新治疗方法的兴趣。三甲基壳聚糖是最近引入的药物载体之一。
设计了阿特里普与三甲基壳聚糖的共轭物，并通过zeta电位仪、原子力显微镜和傅里叶变换红外光谱技术对其进行表征。通过分光光度法测定药物与三甲基壳聚糖的共轭以及纳米共轭物的细胞摄取。采用XTT试验测量细胞毒性。通过酶联免疫吸附测定试验研究抗逆转录病毒效率。
zeta电位仪结果证明，纳米共轭物颗粒团聚的平均尺寸为493.4±24.6nm，但大多数颗粒的尺寸为177.2±7.8nm，强度为87.9%。原子力显微镜技术显示，纳米共轭物和三甲基壳聚糖的尺寸分别为129nm和59.78nm。纳米共轭物的zeta电位为-1.35±0.04mv，药物的zeta电位为-7.69±0.3mv。阿特里普与三甲基壳聚糖的共轭效率为5.27%。在检测浓度下，通过分光光度法测得的纳米共轭物的细胞摄取量约为游离药物的两倍（P=0.007）。与阿特里普相比，纳米共轭物对HIV复制表现出更高的抑制作用（P=0.0001）。
结果表明，阿特里普-三甲基壳聚糖共轭物没有显著的细胞毒性作用。由于纳米共轭物对病毒复制具有更高的抑制作用，它可用于较低浓度的抗病毒治疗，从而降低耐药性和其他副作用。
