负载 ZIF-8的GelMA 水凝胶制备及其药物缓释性能和抑菌作用评价

doi:10.13481/j.1671-587X.20240113

摘要/Abstract

摘要：

目的制备一种含沸石咪唑类骨架材料8（ZIF-8）的复合光交联水凝胶，评价其体外细胞毒性、药物释放能力和抗菌性能。方法采用水热法合成 ZIF-8 颗粒，扫描电子显微镜（SEM）观察 ZIF-8 颗粒微观形态表现。将 ZIF-8 颗粒以质量分数 0.2%的比例与甲基丙烯酸酐化明胶（GelMA）混合获得复合水凝胶 GelMA-Z。采用原子吸收光谱法检测各时间点 GelMA-Z 中锌离子（Zn²⁺）累计释放量。NIH-3T3 细胞分为对照组、GelMA 组和GelMA-Z组，将GelMA和GelMA-Z与 NIH-3T3 细胞共培养 1、3和 7 d，采用 CCK-8 法检测各组细胞存活率。大肠杆菌（E. coli）和金黄色葡萄球菌（S. aureus）分为对照组、GelMA 组和GelMA-Z组，将GelMA和GelMA-Z 分别与E.coli 和S. aureus 共培养6、12和 24 h，采用酶标仪检测不同时间点各组细菌活性，采用平板抑菌实验和活/死细菌染色实验检测各组细菌菌落形成情况和活/死细菌染色情况。结果 SEM 观察，采用水热法合成的 ZIF-8 颗粒具有均匀的粒径。原子吸收光谱法检测，GelMA-Z 中 Zn²⁺在 1 d 内呈突释阶段后缓慢释放，7 d 左右达到平衡。与对照组比较，1、3和 7 d 时GelMA 组和 GelMA-Z 组细胞存活率均大于 90%，差异无统计学意义（P>0.05）。酶标仪检测菌液活性，与细菌共培养 6、12和 24 h时，与对照组和 GelMA 组比较，GelMA-Z 组 E.coli 和 S.aureus 活性明显降低（P<0.05）。平板抑菌实验， GelMA-Z 组中菌液涂布后形成的菌落数明显少于对照组和 GelMA 组。活/死细菌染色实验，GelMA-Z 组存在大量红色荧光染色的死细菌，对照组和 GelMA 组中则为大量绿色荧光染色的活细菌。结论负载 ZIF-8的 GelMA水凝胶可实现原位光交联，具有良好的 Zn²⁺释放能力和抗菌性，是一种可用于感染伤口治疗的新型水凝胶敷料。

关键词: 含沸石咪唑类骨架材料8, 甲基丙烯酸酐化明胶, 水凝胶, 感染伤口, 抗菌性能

Abstract:

objective To prepare a composite photocrosslinked hydrogel containing zeolite imidazole framework-8 （ZIF-8），and to evaluate its in vitro cytotoxicity， drug release capability， and antimicrobial propertie. Methods The ZIF-8 particles were synthesized by hydrothermal method， and the microstructure characteristic was observed under scanning electron microscope （SEM）. The particles were mixed with the gelatin methacryloyl （GelMA） with the mass fraction of 0.2% to obtain the composite hydrogel GelMA-Z. The atomic absorption spectroscope was used to detect the cumulative zinc ion （Zn²⁺） release amounts in GelMA-Z at different time points.The NIH-3T3 cells were co-cultured with GelMA-Z for 1， 3， and 7 d；the viabilities of the cells in various groups were detected by CCK-8 assay； the GelMA-Z was co-cultured with Escherichia coli （E. coli） and Staphylococcus aureus （S. aureus） for 6，12，and 24 h and divided into control group， GelMA group， and GelMA-Z group.The bacterial activities of the cells in various groups at different time points were detected by microplate reader； the bacterial formation and the presence of live/dead becterial staining condition were detected by plate antibacterial experiment and live/dead bacterial staining method. Results The SEM observation results showed that the hydrothermally synthesized ZIF-8 particles had the uniform particle sizes.The atomic absorption spectroscope results showed that Zn²⁺ in GelMA-Z showed an initial burst phase within 1 d， followed by a slow release，and reached the equilibrium around 7 d.Compared with control group，the viabilities the cells in GelMA group and GelMA-Z group were above 90% on the 1st， 3rd， and 7th days， but there was no significant difference（P>0.05）.The bacterial activity detection results showed that when co-cultured with bacteria for 6，12，and 24 h， compared with control group and GelMA group，the bacterial activities of the E. coli and S. aureus in GelMA-Z group were decreased （P<0.05）. The plate antibacterial experiment results showed that the number of bacterial formation in GelMA-Z group was fewer than those in control group and GelMA group. The live/dead bacterial staining results showed that in GelMA-Z group，there was a large number of red fluorescence stained dead bacteria； in control group and GelMA group， there was a large number of green fluorescence stained live bacteria. Conclusion The GelMA hydrogel loaded with ZIF-8 particles can achieve the in situ photocrosslinking and possesses good Zn²⁺ release capability and antimicrobial activity， and it is a novel hydrogel dressing for treatment of the infected wounds.

Key words: Zeolite imidazole framework-8, Gelatin methacryloyl, Hydrogel, Infected wound, Antibacterial property

中图分类号:

R318.08

姜孔昭,李驰宇,罗云纲,刘志辉. 负载 ZIF-8的GelMA 水凝胶制备及其药物缓释性能和抑菌作用评价[J]. 吉林大学学报(医学版), 2024, 50(1): 106-112.

Kongzhao JIANG,Chiyu LI,Yungang LUO,Zhihui LIU. Preparationof GelMA hydrogel loaded with ZIF-8 and evaluation of drug sustained release and antibacterial effect[J]. Journal of Jilin University(Medicine Edition), 2024, 50(1): 106-112.

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Time	Cumulative release amount of Zn²⁺
0 h	0
12 h	0.55±0.03
1 d	0.91±0.02
3 d	1.07±0.02
7 d	1.19±0.03

Group	Survival rate
Group	（t/d） 1	3	7
Control	100.00±0.00	100.00±0.00	100.00±0.00
GelMA	98.21±0.26	98.26±0.49	99.11±0.51
GelMA-Z	97.23±1.17	97.33±0.89	98.75±0.55

Group	Activity of E. coli
Group	（t/h） 0	6	12	24
Control	0.23±0.01	0.72±0.03	0.95±0.08	1.02±0.06
GelMA	0.23±0.01	0.71±0.03	0.96±0.06	1.01±0.07
GelMA-Z	0.23±0.01	0.24±0.01^*△	0.25±0.02^*△	0.24±0.02^*△

Group	Activity of S. aureus
Group	（t/h） 0	6	12	24
Control	0.24±0.02	0.76±0.03	1.02±0.07	1.13±0.03
GelMA	0.24±0.02	0.77±0.03	1.03±0.04	1.12±0.08
GelMA-Z	0.24±0.02	0.25±0.02^*△	0.25±0.05^*△	0.24±0.04^*△

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