负载EGCG和葡聚糖水解酶的纳米囊泡构建及其对变形链球菌抗菌作用的实验研究

doi:10.3969/j.issn.1005-4979.2023.02.002

《口腔颌面外科杂志》 ›› 2023, Vol. 33 ›› Issue (2): 71-76. doi: 10.3969/j.issn.1005-4979.2023.02.002

负载EGCG和葡聚糖水解酶的纳米囊泡构建及其对变形链球菌抗菌作用的实验研究

翟子惠(), 苏俭生()

上海牙组织修复与再生工程技术研究中心，同济大学口腔医学院，同济大学附属口腔医院口腔修复科，上海 200072

修回日期:2023-03-05 接受日期:2022-04-27 出版日期:2023-04-28 发布日期:2023-05-05
通讯作者: 苏俭生，教授. E-mail：sjs@tongji.edu.cn
作者简介:
翟子惠，硕士. E-mail：1931263@tongji.edu.cn
基金资助:
上海市科学技术委员会项目(18441902100); 上海市科学技术委员会项目(201409006200)

An experimental study of construction of EGCG and dextranase loaded nanovesicles and its antibacterial effects on Streptococcus mutans

ZHAI Zihui(), SU Jiansheng()

Shanghai Engineering Research Center of Tooth Restoration and Regeneration, Department of Prosthodontics, School and Hospital of Stomatology, Tongji University, Shanghai 200072, China

Revised:2023-03-05 Accepted:2022-04-27 Online:2023-04-28 Published:2023-05-05

摘要/Abstract

摘要：

目的：探究负载表没食子儿茶素没食子酸酯（epigallocatechin gallate，EGCG）和葡聚糖水解酶的纳米囊泡对变形链球菌(Streptococcus mutans，S.mutans）的抗菌作用。方法：制备负载EGCG和葡聚糖水解酶的纳米囊泡，并利用透射电子显微镜（transmission electron microscope，TEM）对其进行表征，通过紫外吸光度法测定其载药率，通过抑菌及抗菌实验检测其对S.mutans的抗菌效果。结果：负载EGCG和葡聚糖水解酶的纳米囊泡在TEM下为均匀球体，粒径约为100 nm，体外稳定性良好，载药率达74.06%；最小抑菌浓度（minimum inhibitory concentration，MIC）为24 μg/mL，最小杀菌浓度（minimum bactericidal concentration，MBC）为32 μg/mL，显示出抗S.mutans的特性。结论：本实验成功构建了目标纳米囊泡，且其对S.mutans具有抗菌效果。

关键词: 纳米囊泡, 表没食子儿茶素没食子酸酯, 葡聚糖水解酶, 抗菌作用

Abstract:

Objective: To explore the antibacterial effects of a nanovesicle carrier loaded with epigallocatechin gallate (EGCG) and dextranase on Streptococcus mutans (S.mutans). Methods: The nanovesicles loaded with EGCG and dextranase were fabricated and characterized by transmission electron microscopy (TEM). Drug loading rate was determined by ultraviolet spectrophotometry. Antibacterial properties of nanovesicles against S.mutans were evaluated by antibacterial and bacteriostatic tests. Results: The nanovesicles were successfully constructed, which appeared as uniform spheres under TEM. The particle size was nearly 100 nm, the stability in vitro was fair, and the drug loading rate was 74.06%.The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the synthesized nanovesicles was 24 μg/mL and 32 μg/mL respectively, showing its antibacterial effect on S.mutans. Conclusion: The nanovesicles were successfully constructed and had antibacterial effect on S.mutans.

Key words: nanovesicles, epigallocatechin gallate, dextranase, antibacterial effect

中图分类号:

R782

翟子惠, 苏俭生. 负载EGCG和葡聚糖水解酶的纳米囊泡构建及其对变形链球菌抗菌作用的实验研究[J]. 《口腔颌面外科杂志》, 2023, 33(2): 71-76.

ZHAI Zihui, SU Jiansheng. An experimental study of construction of EGCG and dextranase loaded nanovesicles and its antibacterial effects on Streptococcus mutans[J]. 《Journal of Oral and Maxillofacial Surgery》, 2023, 33(2): 71-76.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2023/V33/I2/71

图/表 5

图1 P1的纳米表征 A.P1的TEM结果（×250 000）；B.P1的Zeta电势图；C.粒径分布图。

Figure 1 Nano-characterization of P1

表1 不同投药量对纳米囊泡特征的影响

Table 1 Effects of different dosages of P1 on the characteristics of nanovesicles

PBA-PEI/mg	葡聚糖水解酶/mg	EGCG/mg	分散度	粒径/nm
30 30 30 30 30	10.03 20.06 25.08 28.08 30.09	0.74 1.48 1.85 2.07 2.22	0.474 0.663 0.213 0.202 0.515	021.84 067.91 068.04 100.40 147.20

图2 MTT法测定不同浓度P1溶液的吸光度值 A.培养24 h后；B.培养72 h后。**表示P<0.01, ***表示P<0.001, 与control组比较; NS表示P>0.05,与空白组比较。

Figure 2 The MTT method determined the absorbance value of different concentrations of P1 solution

图3 不同浓度P1溶液中平板菌落分布 A.对照组，P1浓度为0；B.P1浓度为12 μg/mL；C.P1浓度为24 μg/mL；D.P1浓度为32 μg/mL。

Figure 3 Plate colony distribution of different concentrations of P1 solution

图4 培养液在0~32 μg/mL P1浓度下pH值的变化

Figure 4 Changes of pH value of the medium at concentrations of 0-32 μg/mL P1

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负载EGCG和葡聚糖水解酶的纳米囊泡构建及其对变形链球菌抗菌作用的实验研究

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