中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (10): 1551-1554.doi: 10.12307/2022.202

• 材料力学及表面改性 material mechanics and surface modification • 上一篇    下一篇

生物材料表面硬度在巨噬细胞吞噬大肠杆菌生物膜细胞中的作用

赵彦瑞，周君琳   

  1. 首都医科大学附属北京朝阳医院骨科，北京市   100020
  • 收稿日期:2020-09-14 修回日期:2020-09-16 接受日期:2020-12-07 出版日期:2022-04-08 发布日期:2021-10-25
  • 通讯作者: 周君琳，教授，博士生导师，首都医科大学附属北京朝阳医院骨科，北京市 100020
  • 作者简介:赵彦瑞，男，1983年生，河北省承德市人，汉族，主治医师，博士，主要从事创伤方面的研究
  • 基金资助:
    中国留学基金委资助项目(201508110227)

Effect of surface hardness of biomaterials in the phagocytosis of E. coli biofilm cells by macrophages

Zhao Yanrui, Zhou Junlin   

  1. Department of Orthopedics, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China
  • Received:2020-09-14 Revised:2020-09-16 Accepted:2020-12-07 Online:2022-04-08 Published:2021-10-25
  • Contact: Zhou Junlin, Professor, Doctoral supervisor, Department of Orthopedics, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China
  • About author:Zhao Yanrui, Attending physician, MD, Department of Orthopedics, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China
  • Supported by:
    the China Scholarship Council, No. 201508110227 

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摘要： 文题释义：
生物膜细胞：当细菌产生细胞外基质并形成生物膜时，这些感染通常会持续存在。由于细胞外基质的保护和缓慢的生长，生物膜细胞所能耐受的抗生素浓度是同一菌株的浮游细胞的500-5 000倍。 
吞噬作用：作为先天免疫的主要机制，吞噬作用是巨噬细胞针对入侵病原体的受体介导的免疫应答。在吞噬过程中，巨噬细胞利用表面受体识别病原体，将病原体吞噬到质膜来源的吞噬体中，并最终通过多种策略消灭病原体。

背景：医院获得性感染中有60%-70%与不同材料的医疗器械有关，而细菌生物膜细胞在慢性感染中起着重要作用。细菌的黏附和生物膜形成受生物材料特性的影响，例如表面电荷、疏水性、硬度和表面化学性质。
目的：研究巨噬细胞在不同表面硬度生物材料聚二甲基硅氧烷上对大肠杆菌早期生物膜细胞的吞噬作用。
方法：通过控制基料与固化剂的质量比制备硬(5∶1)、中(20∶1)、软(40∶1)3种表面硬度的聚二甲基硅氧烷。①将大肠杆菌接种于3种硬度材料表面，孵育细菌生物膜细胞，将活化的巨噬细胞滴入各材料表面，通过CFU细胞计数、流式细胞仪技术和倒置荧光显微镜检测巨噬细胞在不同表面硬度材料上吞噬大肠杆菌生物膜细胞情况；②将巨噬细胞接种于3种硬度材料表面，随后加入大肠杆菌，CFU细胞计数检测巨噬细胞在不同表面硬度材料上吞噬大肠杆菌生物膜细胞情况。

结果与结论：①先接种细菌后加入巨噬细胞实验：CFU细胞计数、流式细胞仪技术和倒置荧光显微镜检测显示，随着材料表面硬度的增加，巨噬细胞的吞噬作用增强，大肠杆菌生物膜细胞数量减少，3组间细菌生物膜细胞吞噬率比较差异有显著性意义(P < 0.001)；②先接种巨噬细胞后加入细菌实验：CFU细胞计数检测显示，随着材料表面硬度的增加，巨噬细胞的吞噬作用增强，细菌细胞数量减少；显微镜下可见，巨噬细胞在表面硬的材料上更散布、活性更高，并且可见伪足的伸展；③结果表明，随着聚二甲基硅氧烷硬度的增加，巨噬细胞吞噬大肠杆菌生物膜细胞的效率更高，推测通过优化生物材料硬度可以更好地控制内植物相关感染。

https://orcid.org/0000-0002-3957-7419 (赵彦瑞) 


关键词: 材料, 硬度, 聚二甲基硅氧烷, 巨噬细胞, 吞噬作用, 细菌, 生物膜

Abstract: BACKGROUND: Totally 60%-70% of hospital-acquired infections are related to medical devices of different materials, and bacterial biofilm cells play an important role in chronic infections. Bacterial adhesion and biofilm formation are affected by the characteristics of biological materials, such as surface charge, hydrophobicity, hardness, and surface chemistry.
OBJECTIVE: To study the phagocytosis of macrophages on the early biofilm cells of E. coli on polydimethylsiloxane, a biomaterial with different surface hardness.
METHODS: Hard (5:1), medium (20:1) and soft (40:1) polydimethylsiloxane with three surface hardness was prepared by controlling the mass ratio of base material and curing agent. (1) E. coli were inoculated on the surface of three kinds of hard materials to incubate bacterial biofilm cells. Activated macrophages were dropped onto the surface of each material. CFU cell counting, flow cytometry and inverted fluorescence microscope were used to detect the difference in macrophages Phagocytosis of E. coli biofilm cells on surface hardness materials. (2) Macrophages were inoculated on the surface of three hard materials, and then E. coli was added. CFU cell count was applied to detect macrophages phagocytosis of E. coli biofilm cells on materials with different surface hardness.
RESULTS AND CONCLUSION: (1) Bacteria were inoculated first and then macrophages were added to the experiment: CFU cell count, flow cytometry technology and inverted fluorescence microscope detection showed that as the surface hardness of the material increased, the phagocytosis of macrophages increased and the number of E. coli biofilm cells decreased. The difference in the phagocytic rate of bacterial biofilm cells among the three groups was significant (P < 0.001). (2) Macrophages were inoculated first and then bacteria were added to the experiment: CFU cell count test showed that as the surface hardness of the material increased, the phagocytosis of macrophages increased and the number of bacterial cells decreased. It could be seen under the microscope that the macrophages were on the hard surface material more spread, with higher activity, and visible extension of pseudopods. (3) The results show that with the increase of the hardness of polydimethylsiloxane, the efficiency of macrophages phagocytosis of E. coli biofilm cells is higher. It is speculated that by optimizing the hardness of biological materials, endophyte-related infections can be better controlled.

Key words: materials, hardness, polydimethylsiloxane, macrophages, phagocytosis, bacteria, biofilm

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