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    负载槲皮素纳米粒的壳聚糖/丝胶蛋白水凝胶的制备及性能评价

    Preparation and Performance Evaluation of Chitosan/Sericin Hydrogel Loaded with Quercetin Nanoparticles

      摘要
    • 摘要:
      目的  制备负载槲皮素纳米粒的壳聚糖/丝胶蛋白水凝胶伤口敷料(chitosan/silk sericin hydrogel loaded with quercetin nanoparticles，QuNPs-CS-gel)，并对其进行质量和促创面愈合效果评价。
      方法 采用单乳化溶剂挥发法制备活性氧响应型载槲皮素纳米粒(quercetin nanoparticles，QuNPs)，以粒径、多分散系数(polydispersity index，PDI)、包封率等为评价指标确定处方工艺。以壳聚糖和丝胶蛋白为基质材料，京尼平作为交联剂制备水凝胶(chitosan/silk sericin hydrogel，CS-gel)，再将载药纳米粒包载于水凝胶中得到QuNPs-CS-gel。通过水蒸气透过率、保水率等表征选出最佳配比的水凝胶；利用傅里叶变换红外光谱(fourier transform infrared spectrometer，FTIR)、透射(transmission electron microscope，TEM)以及扫描电镜(scanning electron microscope，SEM)对纳米粒和水凝胶结构与形貌进行表征；借助血液相容性及细胞相容性试验评估其生物组织相容性；采取菌落计数法和动物实验来评价水凝胶的抗菌和促创面愈合作用。
      结果 QuNPs平均粒径为(160.60±0.20)nm，PDI为(0.072±0.059)，包封率为(97.12±0.15)%，载药率为(7.53±0.02)%，在第10 h药物释放完全。CS-gel的综合性能较好的处方为：2%壳聚糖2 mL，2%丝胶蛋白1 mL，0.5%京尼平用量1.50 mg。TEM和SEM结果显示：QuNPs呈较规则的球形，分散性良好；QuNPs-CS-gel呈疏松多孔的网络结构。血液相容性结果显示：实验组的溶血率均<5%，表明QuNPs、CS-gel及QuNPs-CS-gel对红细胞破坏率低，血液相容性良好。细胞相容性结果显示，实验组的细胞存活率均>95%，与对照组无差异，在72 h时，细胞保持活力，几乎无死亡细胞，表明QuNPs、CS-gel及QuNPs-CS-gel具有出色的细胞相容性。抗菌作用结果显示，CS-gel和QuNPs-CS-gel抑菌作用接近100%，而QuNPs抑菌作用>20%，表明水凝胶均有良好的抑菌作用。动物实验结果显示：在第14 天 QuNPs-CS-gel组伤口接近完全愈合，而对照组愈合率仅达到(65.92±1.28)%的闭合，表明水凝胶具有较好的促愈合作用。
      结论 QuNPs-CS-gel具有良好理化性能、生物相容性、抗菌以及促创面愈合作用，有望成为一种理想的伤口敷料。

       

      Abstract:
      OBJECTIVE  To prepare the chitosan/silk sericin hydrogel loaded with quercetin nanoparticles(QuNPs-CS-gel) as a wound dressing and evaluate its quality and ability of promoting wound healing.
      METHODS  Quercetin nanoparticles(QuNPs) responsive to reactive oxygen species were prepared by single emulsification solvent evaporation method. The formulation process was optimized using particle size, polydispersity index(PDI), encapsulation efficiency and so on. Chitosan and silk sericin were used as substrate material, and Genipin was used as crosslinking agent to prepare hydrogel(CS-gel). QuNPs-CS-gel was obtained by coating drug-loaded nanoparticles on the hydrogel. The best proportion of hydrogel was selected by water vapor transmission rate、water retention rate and so on. The structures and morphologies of the nanoparticles and hydrogels were characterized by Fourier transform infrared spectroscopy(FTIR), transmission electron microscope(TEM) and scanning electron microscopy(SEM). Biohistocompatibility was evaluated by blood compatibility and cell compatibility tests. The antibacterial and wound healing effects of hydrogels were evaluated by colony counting and animal experiments.
      RESULTS  For QuNPs, the mean particle size of QuNPs was(160.60±0.20)nm, PDI was(0.072±0.059), encapsulation rate was(97.12±0.15)%, drug loading rate was(7.53±0.02)%, and drug release rate achieved 80% at the 10 h. The comprehensive properties of CS-gel were as follows: 2% chitosan 2 mL, 2% sericin 1 mL, 0.5% Genipine 1.50 mg. TEM and SEM results showed that the shape of QuNPs was regular spherical with good dispersion. QuNPs-CS-gel has a loose and porous network structure. Blood compatibility results showed that the hemolysis rates of the experimental groups were all <5%, indicating that QuNPs, CS-gel and QuNPs-CS-gel had low destruction rates of red blood cells and good blood compatibility. The cell compatibility results showed that the cell survival rates of the experimental groups were >95%, and no difference between the control group and the experimental group. At 72 h, the cells remained active with almost no dead cells, indicating that QuNPs, CS-gel and QuNPs-CS-gel had excellent cell compatibility. The results of antibacterial activity showed that the antibacterial activity of CS-gel and QuNPs-CS-gel was close to 100%, while the antibacterial activity of QuNPs could reach >20%, indicating that both hydrogels had good antibacterial activity. The results of animal experiments showed that the wound of QuNPs-CS-gel group was close to complete healing on the 14 day, while the healing rate of the control group was only(65.92±1.28)%, indicating that hydrogel had a better effect on promoting healing.
      CONCLUSION  QuNPs-CS-gel has good properties of characterization, biocompatibility, antibacterial and wound healing, and is expected to be an ideal wound dressing.

       

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