Chinese Journal of Tissue Engineering Research ›› 2024, Vol. 28 ›› Issue (15): 2330-2337.doi: 10.12307/2024.377
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Guo Yuxin, Wang Hao, Li Mingqi, Chen Yueying, Pan Juhong, Huang Xin, Wang Zhiwen, Zhou Qing
Received:
2023-05-11
Accepted:
2023-06-25
Online:
2024-05-28
Published:
2023-09-19
Contact:
Zhou Qing, PhD, Professor, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China
About author:
Guo Yuxin, Master candidate, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China
Supported by:
CLC Number:
Guo Yuxin, Wang Hao, Li Mingqi, Chen Yueying, Pan Juhong, Huang Xin, Wang Zhiwen, Zhou Qing. Ultrasound-optimized hydrogel scaffold used to promote transdermal delivery of gold nanoparticles[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(15): 2330-2337.
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2.5 AuNPs-NDs和AuNPs-NDs@gel的生物相容性评价 将L929细胞分别与不同质量浓度的AuNPs-NDs孵育12 h和24 h,细胞存活率均> 90%。在相同AuNPs-NDs质量浓度条件下,孵育12 h与孵育24 h之后的细胞存活率比较差异无显著性意义(P > 0.05),当AuNPs-NDs质量浓度达6.0 mg/mL时,孵育24 h与孵育12 h相比细胞活力显著下降(P < 0.05);在相同时间条件下,AuNPs-NDs质量浓度低于1.0 mg/mL时细胞存活率与对照组比较差异无显著性意义(P > 0.05),大于此质量浓度后细胞活性开始下降,但总体存活率均> 85%,见图6A、B。 水凝胶支架的细胞相容性通过Live/Dead染色观察L929细胞的生长活性进行评价。培养24 h后,如图6C-K结果所示,水凝胶支架及其降解产物对L929细胞没有显示出明显的毒性,并且低强度脉冲超声+AuNPs-NDs@gel组还表现出更少的凋亡细胞。"
2.6 水凝胶支架的形貌观察 扫描电镜观察PDA@GelMA和AuNPs-NDs@gel的微观结构,见图7。两种材料截面均为多孔网状结构,孔隙呈类圆形,AuNPs-NDs@gel还可在较大孔内见到小孔分布。PDA@GelMA和AuNPs-NDs@gel的孔径大小分别为(73.24±16.26),(76.05±16.26) μm,这可能源于交联密度增大导致的孔径减小;定量分析测定PDA@GelMA的孔隙率为(57.6±5.0)%,AuNPs-NDs@gel的孔隙率为(65.7±3.6)%,二者比较差异有显著性意义(P < 0.01),说明低强度脉冲超声能够一定程度上提高水凝胶支架内部的孔隙率。"
图8A显示,随振荡频率从0.1 rad/s增至100 rad/s时,PDA@GelMA和AuNPs-NDs@gel的储能模量(G’)数值都显示出小幅稳定增加,表明水凝胶支架稳定的力学性能。但15% GelMA的G’ 低于PDA@GelMA,这可能源于纯GelMA材料低交联密度所致其较差的力学性能,而PDA@GelMA则由于GelMA与PDA间形成的氢键及π-π键增加了交联密度,表现出更强的力学性能;AuNPs-NDs@gel的G’ 略高于PDA@GelMA,表现出更高的弹性模量,图8C、D展示了PDA@GelMA和AuNPs-NDs@gel良好的组织黏附性和延展性。 适配皮肤的支架材料需具备良好的延展性,因此有必要评估材料的拉伸性能。如图8B所示,AuNPs-NDs@gel相较PDA@GelMA显示出更好的延展性,二者拉伸断裂伸长率分别为122%,106%,优于人体皮肤的60%-70%[14]。AuNPs-NDs的添加使PDA@GelMA表现出更高的拉伸应力,这一结果与流变学相吻合,证明了其更优异的力学性能。 2.8 水凝胶支架的体外凝血性能检测结果 PDA@GelMA和AuNPs-NDs@gel组蒸馏水颜色均未发生明显变化,血液几乎均凝结于水凝胶上,而止血海绵组血块周边可见少量血液漫出,直观说明PDA@GelMA本身的凝血能力优于止血海绵,见图9A。凝血指数可反映材料的凝血性能,其值越小凝血性能越好。图9B是对材料凝血能力的定量分析,止血海绵的凝血指数为(75.3±1.81)%,PDA@GelMA的凝血指数为(40.83±2.06)%,AuNPs-NDs@gel组凝血指数为(37.64±1.16)%,3组间凝血指数两两比较差异有显著性意义(P < 0.01,P < 0.000 1)。"
2.10 水凝胶支架的抗菌性能检测结果 AuNPs-NDs@gel发挥抗菌效能主要源自金纳米粒子的释放。实验组仅有少量菌落生长,对照组对菌落生长抑制不明显,见图11A-C;实验组经超声辐照后对金黄色葡萄球菌的抑菌率平均为83.17%,大于对照组(P < 0.001),见图11D,对照组表现出的抗菌作用可能源于少量未被包载的金纳米粒子。为了更好地评估AuNPs-NDs@gel的杀菌性能,可通过测量600 nm处细菌的吸光度值表征,见图11E。最小抑菌浓度测定结果显示,与PDA@GelMA相比,AuNPs-NDs@gel在超声辐照后的吸光度值显著降低,最小抑菌浓度为 6.5 μg/mL。结果表明AuNPs-NDs@gel具有良好的杀菌效能。"
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