Preparation and inflammation suppression of pH sensitive chitosan/dexamethasone sodium phosphate nano-drug delivery system

doi:10.3969/j.issn.2095-4344.0919

Abstract

Abstract:

BACKGROUND: The clinical utilization of dexamethasone sodium phosphate in the treatment of asthma is limited due to its toxicity and short metabolic time, which means, a slow-release preparation of dexamethasone sodium phosphate is a good solution.

OBJECTIVE: To prepare a pH sensitive chitosan/dexamethasone sodium phosphate nanoparticle, and to investigate its drug loading, microstructure, particle size distribution, in vitro release characteristics, and anti-inflammatory effect.

METHODS: Chitosan as raw material was used to prepare pH sensitive chitosan/dexamethasone sodium phosphate nanoparticles by ionic gelation method. Drug-polymer interactions, morphology, particle size distribution and in vitro release characteristics of the nanoparticles were then detected. Thirty-two Sprague-Dawley rats were randomly divided into four groups: the normal group was subcutaneously injected 1 mL of normal saline at days 0 and 7, and inhaled 1 mL of atomized saline, 30 minutes per day, from day 14 to day 21. The asthma group was subcutaneously injected 1 mL of sensitization solution at days 0 and 7, and inhaled 1 mL of atomized saline solution containing 1% ovalbumin, 30 minutes per day, from day 14 to day 21. The control group was intraperitoneally injected dexamethasone sodium phosphate (0.25 mg/kg) at 30 minutes before induction of asthma, and the other procedures were the same as those used in the asthma group. The experimental group was intraperitoneally injected chitosan/dexamethasone sodium phosphate nanoparticles (0.25 mg/kg dexamethasone sodium phosphate) at 30 minutes before induction of asthma, and the other procedures were the same as those used in the asthma group. Lung tissue samples from each rat were taken at day 14 for histological observation using hematoxylin-eosin staining.

RESULTS AND CONCLUSION: (1) With the increase of dexamethasone sodium phosphate concentration, the drug-loading rate of chitosan/dexamethasone sodium phosphate nanoparticles increased gradually. (2) The chitosan/dexamethasone sodium phosphate nanoparticles obtained were white oval, 95% of which had a diameter of 40-70 nm, with the average diameter being 53.2 nm. (3) In the simulated intestine solution (pH=7.4), sodium dexamethasone phosphate was gradually released from the nanoparticles and completely released within 120 minutes. In the simulated gastric juice (pH=2.1 and pH=4.6), no sodium dexamethasone phosphate was released within 120 minutes. (4) Hematoxylin-eosin staining results showed that the bronchial smooth muscle in the asthma group was hypertrophic accompanied by mucosal hyperemia and edema and infiltration of plenty inflammatory cells. Bronchial inflammation was relieved in the control group, and considerably alleviated in the experimental group, where only a small number of inflammatory cells were infiltrated around the bronchial wall. We make a conclusion from these findings that pH-sensitive chitosan/dexamethasone sodium phosphate nanoparticles exhibit a slow-release potential, which can be used to inhibit the production of inflammatory cells in asthmatic rats given broncho-alveolar lavage

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Chitosan, Inflammation, Asthma, Delayed-Action Preparations, Tissue Engineering

CLC Number:

R318

Chen Na, Xie Tie-min, Zhang Yi-fan, Wang Jiang, Wang Qing-feng. Preparation and inflammation suppression of pH sensitive chitosan/dexamethasone sodium phosphate nano-drug delivery system[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(22): 3550-3556.

Figures/Tables 5

2.1 壳聚糖/地塞米松磷酸钠纳米颗粒载药量的测定随着地塞米松磷酸钠加入量的增加，纳米颗粒的载药量也随之增加。重复数为10的条件下，当地塞米松磷酸钠的质量浓度依次为4，5，6 g/L时，地塞米松磷酸钠的载药量依次递增，分别为(24.13±1.29)%，(31.39±2.21)%，(36.98± 1.45)%，组间两两比较差异有显著性意义(P < 0.05)，这与Pignatello等[28]的文章结论是一致的。由于地塞米松磷酸钠溶液的pH为6.0，与地塞米松磷酸钠的解离常数pK2相近[29]，而地塞米松磷酸钠电离为带两个负电荷的离子，可通过静电作用结合壳聚糖质子化的NH3+ [30]，在多聚磷酸钠的交联下形成纳米颗粒。另一方面，随着地塞米松磷酸钠质量浓度的递增，壳聚糖/地塞米松磷酸钠的载药量增长趋势却并未呈线性，如当地塞米松磷酸钠的质量浓度由4 g/L上升至5 g/L时，壳聚糖/地塞米松磷酸钠的载药量上升了近30%；当地塞米松磷酸钠的质量浓度由5 g/L上升至6 g/L时，壳聚糖/地塞米松磷酸钠的载药量上升了近17%，该结果说明当地塞米松磷酸钠的质量浓度为5 g/L时，该颗粒的载药量已达到峰值。 2.2 壳聚糖/地塞米松磷酸钠纳米颗粒的红外光谱表征结果图3展示了地塞米松磷酸钠、壳聚糖/地塞米松磷酸钠和壳聚糖的红外光谱图，在1 458 cm-1、1 380 cm-1处的特征峰是P=O的振动引起的，并且在1 722 cm-1和1 644 cm-1的特征吸是C=O的振动引起的，而1 620 cm-1特征峰是C=C伸缩振动引起的，这几个特征峰体现了地塞米松磷酸钠的分子结构特征[27]；在2 390 cm-1、1 090 cm-1处的特征峰是由地塞米松磷酸钠和壳聚糖分子结构中的-CH3和-C-O振动引起的；1 533 cm-1特征峰是壳聚糖分子结构中的-NH3+基团的伸缩振动引起的[31]。以上结果说明，地塞米松磷酸钠被包裹在壳聚糖/地塞米松磷酸钠纳米颗粒中。 2.3 壳聚糖/地塞米松磷酸钠纳米颗粒微观表征结果壳聚糖与地塞米松磷酸钠质量浓度比为1∶1制得相应的纳米颗粒，其形貌特征如图4所示，可以看出壳聚糖/地塞米松磷酸钠纳米颗粒为白色椭球形，大小约为50 nm。进一步采用激光粒度对所制得颗粒的粒径进行分析，如图5所示：95%的纳米颗粒直径在40-70 nm之间，平均粒径为 54.4 nm。 "

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