Abstract：
OBJECTIVE: Trinitroglycerin (TNG) is a remarkable NO-releasing agent. Here, we synthesized TNG based on chitosan Nanogels (Ngs) for ameliorating complications associated with high-dose TNG administration.
METHODS: TNG-Ngs fabricated through ionic-gelation technique. Fourier-transformed infrared (FT-IR), zeta-potential, dynamic light scattering (DLS), and electron microscopy techniques evaluated the physicochemical properties of TNG-Ngs. MTT was used to assess the biocompatibility of TNG-Ngs, as the antioxidative properties were determined via lactate dehydrogenase (LDH), reactive oxygen species (ROS), and lipid peroxide (LPO) assays. The antibacterial activity was evaluated against Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), Methicillin-resistant Staphylococcus aureus (MRSA), and vancomycin-resistant Enterococci (VRE).
RESULTS: Physicochemical characterization reveals that TNG-Ngs with size diameter (96.2 ± 29 nm), polydispersity index (PDI, 0.732), and negative zeta potential (-1.1 mv) were fabricated. The encapsulation efficacy (EE) and loading capacity (LC) were obtained at 71.1 % and 2.3 %, respectively, with no considerable effect on particle size and morphology. The cytotoxicity assay demonstrated that HepG2 cells exposed to TNG-Ngs showed relative cell viability (RCV) of >80 % for 70 μg/ml compared to the TNG-free drug at the same concentration (P < 0.05). TNG-Ngs showed significant differences with the TNG-free drug for LDH, LPO, and ROS formation at the same concentration (P < 0.001). The antibacterial activity of the TNG-Ngs against S. aureus, E. coli, VRE, and MRSA was higher than the TNG-free drug and Ngs (P < 0.05).
CONCLUSIONS: TNG-Ngs with enhanced antibacterial and antioxidative activity and no obvious cytotoxicity might be afforded as novel nanoformulation for promoting NO-dependent diseases.
METHODS: TNG-Ngs fabricated through ionic-gelation technique. Fourier-transformed infrared (FT-IR), zeta-potential, dynamic light scattering (DLS), and electron microscopy techniques evaluated the physicochemical properties of TNG-Ngs. MTT was used to assess the biocompatibility of TNG-Ngs, as the antioxidative properties were determined via lactate dehydrogenase (LDH), reactive oxygen species (ROS), and lipid peroxide (LPO) assays. The antibacterial activity was evaluated against Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), Methicillin-resistant Staphylococcus aureus (MRSA), and vancomycin-resistant Enterococci (VRE).
RESULTS: Physicochemical characterization reveals that TNG-Ngs with size diameter (96.2 ± 29 nm), polydispersity index (PDI, 0.732), and negative zeta potential (-1.1 mv) were fabricated. The encapsulation efficacy (EE) and loading capacity (LC) were obtained at 71.1 % and 2.3 %, respectively, with no considerable effect on particle size and morphology. The cytotoxicity assay demonstrated that HepG2 cells exposed to TNG-Ngs showed relative cell viability (RCV) of >80 % for 70 μg/ml compared to the TNG-free drug at the same concentration (P < 0.05). TNG-Ngs showed significant differences with the TNG-free drug for LDH, LPO, and ROS formation at the same concentration (P < 0.001). The antibacterial activity of the TNG-Ngs against S. aureus, E. coli, VRE, and MRSA was higher than the TNG-free drug and Ngs (P < 0.05).
CONCLUSIONS: TNG-Ngs with enhanced antibacterial and antioxidative activity and no obvious cytotoxicity might be afforded as novel nanoformulation for promoting NO-dependent diseases.
摘要：
目的:三硝基甘油(TNG)是一种显著的NO释放剂。这里,我们合成了基于壳聚糖纳米凝胶（Ngs）的TNG，用于改善与高剂量TNG给药相关的并发症。
方法:通过离子凝胶化技术制备TNG-Ngs。傅里叶变换红外(FT-IR),zeta电位,动态光散射(DLS),和电子显微镜技术评估了TNG-Ngs的物理化学性质。MTT用于评估TNG-Ngs的生物相容性，由于抗氧化特性是通过乳酸脱氢酶（LDH）测定的，活性氧（ROS），和脂质过氧化物(LPO)测定。对金黄色葡萄球菌(S.金黄色葡萄球菌)，大肠杆菌(E.大肠杆菌)，耐甲氧西林金黄色葡萄球菌(MRSA),耐万古霉素肠球菌（VRE）。
结果：物理化学表征表明，TNG-Ngs的尺寸直径（96.2±29nm），多分散指数(PDI，0.732),和负zeta电位（-1.1mv）被制造。包封效率(EE)和负载能力(LC)分别为71.1%和2.3%,分别,对颗粒尺寸和形态没有显著影响。细胞毒性测定表明,与相同浓度的不含TNG的药物相比,暴露于TNG-Ngs的HepG2细胞对于70μg/ml显示>80%的相对细胞活力(RCV)(P<0.05)。TNG-Ngs与不含TNG的LDH药物有显著差异，LPO,在相同浓度下形成ROS(P<0.001)。TNG-Ngs对金黄色葡萄球菌的抗菌活性，大肠杆菌，VRE,MRSA高于无TNG药物和Ngs(P<0.05)。
结论：TNG-Ngs具有增强的抗菌和抗氧化活性，并且没有明显的细胞毒性，可能作为促进NO依赖性疾病的新型纳米制剂提供。
方法:通过离子凝胶化技术制备TNG-Ngs。傅里叶变换红外(FT-IR),zeta电位,动态光散射(DLS),和电子显微镜技术评估了TNG-Ngs的物理化学性质。MTT用于评估TNG-Ngs的生物相容性，由于抗氧化特性是通过乳酸脱氢酶（LDH）测定的，活性氧（ROS），和脂质过氧化物(LPO)测定。对金黄色葡萄球菌(S.金黄色葡萄球菌)，大肠杆菌(E.大肠杆菌)，耐甲氧西林金黄色葡萄球菌(MRSA),耐万古霉素肠球菌（VRE）。
结果：物理化学表征表明，TNG-Ngs的尺寸直径（96.2±29nm），多分散指数(PDI，0.732),和负zeta电位（-1.1mv）被制造。包封效率(EE)和负载能力(LC)分别为71.1%和2.3%,分别,对颗粒尺寸和形态没有显著影响。细胞毒性测定表明,与相同浓度的不含TNG的药物相比,暴露于TNG-Ngs的HepG2细胞对于70μg/ml显示>80%的相对细胞活力(RCV)(P<0.05)。TNG-Ngs与不含TNG的LDH药物有显著差异，LPO,在相同浓度下形成ROS(P<0.001)。TNG-Ngs对金黄色葡萄球菌的抗菌活性，大肠杆菌，VRE,MRSA高于无TNG药物和Ngs(P<0.05)。
结论：TNG-Ngs具有增强的抗菌和抗氧化活性，并且没有明显的细胞毒性，可能作为促进NO依赖性疾病的新型纳米制剂提供。
