依诺沙星温敏凝胶的制备及抗菌活性研究

Preparation and antibacterial activity of Enoxacin thermosensitive gel

关键词:  依诺沙星  固体脂质纳米粒  温敏凝胶  抗菌活性

Key words:  Enoxacin  solid lipid nanoparticles  thermosensitive gel  antibacterial activity

摘要:

目的 制备依诺沙星固体脂质纳米粒温敏凝胶（Enoxacin solid lipid nanoparticles gel,ENX-SLN-Gel）并考察其体外抗菌活性。方法 采用乳化蒸发-低温固化法制备依诺沙星固体脂质纳米粒，以包封率、载药量为考察指标，通过单因素实验和Box-Behnken响应面法优化制备工艺，以粒径、Zeta电位、差示扫描量热法、傅里叶变换红外光谱及X射线衍射法对其进行表征；采用冷溶法制备温敏凝胶并进行质量评价；通过CCK8法考察细胞毒性并进行皮肤刺激性实验观察其皮肤用药安全性；抑菌圈实验比较ENX和ENX-SLN-Gel的体外抗菌活性。结果 优化后的ENX-SLN的包封率为（89.86±0.60），载药量为（4.20±0.03），平均粒径为（262.4±0.72）nm，PDI为0.167±0.64，Zeta电位为（-14.3±5.25），DSC、FT-IR、XRD进一步验证了ENX-SLN的形成；所得凝胶的胶凝温度为（32.4±0.46）℃，pH值为6.30±0.13；ENX-SLN在250 μg·mL-1浓度范围内时，细胞的存活率为95 %以上；皮肤刺激性实验显示，完整皮肤和破损皮肤组均未见红斑、水肿等刺激性反应，说明其安全性良好；ENX和ENX-SLN-Gel的抑菌圈直径分别为（9.6±0.7）mm,（14.5±1.0）mm。结论 ENX-SLN-Gel制备工艺方法简单，合理可行，具有良好的抗菌效果，有望成为外用制剂的一种新途径。

Abstract:

OBJECTIVE Enoxacin solid lipid nanoparticles gel (ENX-SLN Gel) was prepared and its antibacterial activity in vitro was investigated. METHODS Enoxacin solid lipid nanoparticles were prepared by emulsion evaporation low-temperature solidification method, with encapsulation efficiency and drug loading as evaluation indicators. The preparation process was optimized through single factor experiments and Box Behnken response surface methodology. The particles were characterized by particle size, polydispersity index, Zeta potential, differential scanning calorimetry, Fourier transform infrared spectroscopy, and X-ray diffraction; Temperature sensitive gel was prepared by cold solution method and its quality was evaluated; Using CCK8 method to investigate cell toxicity and conducting skin irritation experiments to observe the safety of skin medication; Compare the in vitro antibacterial activity of ENX and ENX SLN Gel through inhibition zone experiments. RESULTS The encapsulation efficiency of the optimized ENX-SLN was (89.86±0.60), the drug loading was (4.20±0.03), the average particle size was (262.4±0.72) nm, the PDI was 0.167±0.64, and the Zeta potential was (-14.3±5.25). DSC, FT-IR, and XRD further confirmed the formation of ENX-SLN; The gelling temperature of the obtained gel is (32.4±0.46) ℃, and the pH value is 6.30±0.13; When the concentration range of ENX-SLN is 250 μg·mL-1, the cell survival rate is over 95 %; The skin irritation test showed that there were no irritating reactions such as erythema or edema observed in both intact and damaged skin groups, indicating good safety; The diameters of the inhibition zones for ENX and ENX SLN Gel are (9.6±0.7) mm and (14.5 ±1.0) mm, respectively. CONCLUSION The preparation process of ENX-SLN Gel is simple, reasonable and feasible, with good antibacterial effects, and is expected to become a new approach for topical formulations.