葛根素纳米银合成及光热杀菌和促糖尿病感染伤口愈合作用研究

doi:10.6023/A24080252

化学学报 ›› 2024, Vol. 82 ›› Issue (11): 1150-1161.DOI: 10.6023/A24080252 上一篇下一篇

研究论文

葛根素纳米银合成及光热杀菌和促糖尿病感染伤口愈合作用研究

王英辉^a, 王雨辉^b, 熊佳瑶^b, 苏思琪^b, 郝梦珂^b, 魏思敏^b^,^*()

a 长安大学理学院陕西西安 710064
b 陕西中医药大学陕西中药资源产业化省部共建协同创新中心秦药特色资源研究开发国家重点实验室(培育) 陕西咸阳 712083

投稿日期:2024-08-25 发布日期:2024-10-22
基金资助:
国家自然科学基金(22103007); 国家自然科学基金(82274353); 陕西省重点研发计划(2024SF-YBXM-517); 陕西省重点研发计划(2024SF-YBXM-426); 陕西省创新能力支撑计划-青年科技新星项目(2023KJXX-063); 中央高校基本科研业务费(300102124207); 陕西中医药大学2023年度科技创新人才体系建设计划(2023-LJRC-03); 秦药特色资源研究开发重点实验室开放课题项目(SUCM-QM202207)

The Fabrication of Silver Nanoparticles Using Puerarin and the Photothermal Sterilization and Diabetic Wound Healing Behavior

Yinghui Wang^a, Yuhui Wang^b, Jiayao Xiong^b, Siqi Su^b, Mengke Hao^b, Simin Wei^b()

a College of Science, Chang’an University, Xi’an 710064, China
b State Key Laboratory of Research & Development of Characteristic Qin Medicine Resources (Cultivation), Co-construction Collaborative Innovation Center for Chinese Medicine Resources Industrialization by Shaanxi & Education Ministry, Shaanxi University of Chinese Medicine, Xianyang 712083, China

Received:2024-08-25 Published:2024-10-22
Contact: ^*E-mail: weismiccas@163.com
Supported by:
National Natural Science Foundation of China(22103007); National Natural Science Foundation of China(82274353); Key Research and Development Plan of Shaanxi Province(2024SF-YBXM-517); Key Research and Development Plan of Shaanxi Province(2024SF-YBXM-426); Science and Technology Youth Stars Project of Shaanxi Province(2023KJXX-063); Fundamental Research Funds for the Central Universities, CHD(300102124207); Science and Technology Innovation Talent System Construction Plan of Shaanxi University of Chinese medicine(2023-LJRC-03); Open Project of State Key Laboratory of Research and Development of Characteristic Qin Medicine Resources(SUCM-QM202207)

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1. Supporting Information-A24080252.pdf(359KB)

摘要/Abstract

本工作直接使用天然药物分子葛根素作为还原剂和保护剂绿色制备了纳米银(Pue@AgNPs). Pue@AgNPs为近球形, 粒径主要分布在10~20 nm, 有少量聚集体, 表面吸附带负电荷物质, 为面心立方结构. Pue@AgNPs具有光热效应, 可捕获2,2'-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐自由基阳离子(ABTS^•+)和1,1-二苯基-2-三硝基苯肼(DPPH)自由基, 被用作光热试剂和抗氧化剂. 对大肠杆菌(E. coli)、金黄色葡萄球菌(S. aureus)和耐甲氧西林金黄色葡萄球菌(MRSA)最小抑制浓度均为50.0 μg•mL⁻¹; 可通过减轻伤口处炎症反应、加速新生血管和胶原纤维生成促进糖尿病大鼠伤口闭合.

关键词: 葛根素, 纳米银, 光热效应, 抑菌剂, 抗氧化剂, 伤口敷料

The complexity of plant extract incurs the uncertainty of capping and reducing agents in the biosynthesis of silver nanoparticles (AgNPs) leading to a poor reproducibility, which dramatically limits the application of this method. To overcome this problem, the puerarin (Pue), which is an isoflavone derived from kudzu roots and has strong biological activities, is directly used as the reducing and capping agents to prepare bioactive AgNPs (Pue@AgNPs). The influence of biosynthesis parameters like pH (Pue solution), concentration of both Pue and AgNO₃ and incubation time were assessed to obtain Pue@AgNPs with superior bactericidal activity and photothermal effect. The as-prepared AgNPs are nearly spherical mainly existing in the monodispersed form with a small amount of aggregation, covering by anions (－44.0 mV) and revealing a high degree of stability (at least 15 days). It is conceived that the aggregation of Pue@AgNPs is responsible for absorbing near-infrared light (808 nm). The X-ray diﬀraction (XRD), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM) and selected area electron diﬀraction (SAED) studies show that Pue@AgNPs has a face-centered cubic (fcc) structure. The Pue@AgNPs show potent photothermal effect and antibacterial activity against Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) and Methicillin-resistant Staphylococcus aureus (MRSA) under 808 nm laser radiation. The minimum inhibition concentration (MIC) values for E. coli, S. aureus and MRSA are 50.0 μg•mL⁻¹. The internalization of Pue@AgNPs displays a variant trend suggesting the diﬀerent mechanism of cell death. For E. coli, cell wall and intracellular damage may be responsible for cell death. However, for S. aureus and MRSA, the cell death may be rooted in oxidative stress or intracellular penetration. The Pue@AgNPs also exhibits superior antioxidation activity in trapping 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radical cation (ABTS^•+) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, and could accelerate diabetic wound contraction and healing within 21 days. The histological analysis showed that Pue@AgNPs could reduce inflammation and accelerate the formation of blood and collagen deposition.

Key words: puerarin, silver nanoparticles, photothermal effect, antibacterial agent, antioxidant, wound dressing

引用此文

王英辉, 王雨辉, 熊佳瑶, 苏思琪, 郝梦珂, 魏思敏. 葛根素纳米银合成及光热杀菌和促糖尿病感染伤口愈合作用研究[J]. 化学学报, 2024, 82(11): 1150-1161.

Yinghui Wang, Yuhui Wang, Jiayao Xiong, Siqi Su, Mengke Hao, Simin Wei. The Fabrication of Silver Nanoparticles Using Puerarin and the Photothermal Sterilization and Diabetic Wound Healing Behavior[J]. Acta Chimica Sinica, 2024, 82(11): 1150-1161.

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图/表 12

图1. 葛根素化学结构式

Figure 1. The structure of puerarin

图2. 不同(A) pH, (B) AgNO3浓度, (C)葛根素浓度及(D)反应时间的紫外-可见吸收光谱

Figure 2. UV-vis absorption spectra of the mixture at different (A) pH, (B) AgNO3 concentration, (C) Puerarin concentration and (D) incubation time

图3. 不同(A) pH, (B)葛根素浓度, (C) AgNO3浓度及(D)反应时间合成的纳米银平均粒径; 不同(E) pH, (F)葛根素浓度, (G) AgNO3浓度及(H)反应时间合成的纳米银多分散指数

Figure 3. The average size of AgNPs at different (A) pH, (B) Puerarin concentration, (C) AgNO3 concentration and (D) incubation time; the PDI of AgNPs at different (E) pH, (F) Puerarin concentration, (G) AgNO3 concentration and (H) incubation time

图4. (A) EDX光谱; (B)和(C) TEM图; (D)平均粒径; (E) HRTEM图; (F) XRD图和(G)Zeta电势

Figure 4. (A) EDX spectrum; (B) and (C) TEM images; (D) average size; (E) HRTEM image; (F) XRD spectrum; (G) Zeta potential

图5. 不同AgNPs浓度(A)红外热成像图; (B)光热曲线; 不同激光强度照射AgNPs后(C)红外热成像图; (D)光热曲线

Figure 5. (A) Infrared thermal images; (B) Photothermal heating curves of AgNPs with different concentrations; (C) infrared thermal images; (D) photothermal heating curves of AgNPs with di?erent laser power densities

图6. 无808 nm激光辐射时AgNPs对(A) E. coli, (B) S. aureus和(C) MRSA的抑制作用; 808 nm激光辐射时(D) E. coli, (E) S. aureus和(F) MRSA的抑制作用

Figure 6. The antimicrobial activity of AgNPs against (A) E. coli, (B) S. aureus and (C) MRSA without 808 nm laser radiation; the antimicrobial activity of AgNPs against (D) E. coli, (E) S. aureus and (F) MRSA with 808 nm laser radiation

图7. (A) E. coli, (B) S. aureus和(C) MRSA在固体培养基上生长的照片

Figure 7. Photographs of (A) E. coli, (B) S. aureus and (C) MRSA colonies grown on LB agar plates

图8. 无808 nm激光照射时, (A) E. coli, (C) S. aureus和(E) MRSA对AgNPs摄入分析; 808 nm激光照射时, (B) E. coli, (D) S. aureus和(F) MRSA对AgNPs摄入分析

Figure 8. AgNPs uptake analysis in (A) E. coli, (C) S. aureus and (E) MRSA without 808 nm laser radiation; AgNPs uptake analysis in (B) E. coli, (D) S. aureus and (F) MRSA with 808 nm laser radiation

图9. 无808 nm激光照射时, (A) E. coli, (B) S. aureus和(C) MRSA的TEM图; 808 nm激光照射时, (D) E. coli, (E) S. aureus和(F) MRSA的TEM图

Figure 9. TEM images of (A) E. coli, (B) S. aureus and (C) MRSA without 808 nm laser radiation; TEM images of (D) E. coli, (E) S. aureus and (F) MRSA with 808 nm laser radiation

图10. AgNPs对(A) ABTS?+, (B) DPPH自由基和(C) ?OH的捕获率

Figure 10. Free radical scavenging rate of AgNPs against (A) ABTS?+, (B) DPPH radical and (C) ?OH

图11. AgNPs处理伤口不同时间后的图片

Figure 11. The pictures of the wound area at di?erent time of AgNPs treatment groups

图12. 不同浓度AgNPs处理21天后(A) H&E和(B) Masson染色图片

Figure 12. Images of (A) H&E; (B) Masson trichromatic staining of wounds with different concentration of AgNPs at day 21

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葛根素纳米银合成及光热杀菌和促糖尿病感染伤口愈合作用研究

The Fabrication of Silver Nanoparticles Using Puerarin and the Photothermal Sterilization and Diabetic Wound Healing Behavior

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