<i>In vivo</i> study on the effects of intercellular adhesion operon of <i>Staphylococcus epidermidis</i> on the inflammation associated with bacteria-fungal mixed biofilm_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANG Guojing ¹ , WAN Zilin ¹ ,  WANG Xiaoyan ¹ , HUANG Yunchao ² , ZHOU Youquan ³ , CHEN Ying ² , LEI Yujie ² , QIAO Limei ¹

1. Department of Cardiovascular Surgery, Yan’an Hospital Affiliated to Kunming Medical University, Kunming Yunnan, 650051, P.R.China;
2. Department of Cardiovascular Thoracic Surgery, the Third Affiliated Hospital of Kunming Medical University, Tumor Hospital of Yunnan Province, Kunming Yunnan, 650118, P.R.China;
3. Department of Clinical Laboratory, the Third Affiliated Hospital of Kunming Medical University, Tumor Hospital of Yunnan Province, Kunming Yunnan, 650118, P.R.China;

Corresponding?author：

WANG Xiaoyan, Email: WANGXY2004@126.com

Keywords：

Biomaterial centered infection; intercellular adhesion operon; Staphylococcus epidermidis; Candida albicans; mixed biofilm; rabbit

DOI：

10.7507/1002-1892.202104061

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To study the effect of intercellular adhesion (ica) operon of Staphylococcus epidermidis on the inflammation associated with mixed biofilm of Staphylococcus epidermidis and Candida albicans on endotracheal tube material in rabbits. Methods The standard strains of Staphylococcus epidermidis RP62A (ica operon positive, positive group) and ATCC12228 (ica operon negative, negative group) were taken to prepare a bacterial solution with a concentration of 1×10⁶ CFU/mL, respectively. Then, the two bacterial solutions were mixed with the standard strain of Candida albicans ATCC10231 of the same concentration to prepare a mixed culture solution at a ratio of 1∶1, respectively. The mixed culture solution was incubated with endotracheal tube material for 24 hours. The formation of mixed biofilm on the surface of the material was observed by scanning electron microscope. Thirty New Zealand rabbits, aged 4-6 months, were divided into two groups (n=15), and the endotracheal tube materials of the positive group and the negative group that were incubated for 24 hours were implanted beside the trachea. The body mass of rabbits in the two groups was measured before operation and at 1, 3, and 7 days after operation. At 1, 3, and 7 days after operation, the levels of interleukin 1β (IL-1β), IL-6, tumor necrosis factor α (TNF-α), and monocytechemotactic protein 1 (MCP-1) were detected by using an ELISA test kit. At 7 days after operation, the formation of mixed biofilm on the surface of the endotracheal tube materials was observed by scanning electron microscope, the inflammation and infiltration of tissues around the materials were observed by HE staining, and the bacterial infections in heart, lung, liver, and kidney were observed by plate colony counting method.Results Scanning electron microscope observation showed that the mixed biofilm structure was obvious in the positive group after 24 hours in vitro incubation, but no mixed biofilm formation was observed in the negative group. In vivo studies showed that there was no significant difference in body mass between the two groups before operation and at 1, 3, and 7 days after operation (P>0.05). Compared with the negative group, the levels of MCP-1 and IL-1β at 1 day, and the levels of IL-1β, MCP-1, IL-6, and TNF-α at 3 and 7 days in the positive group all increased, with significant differences (P<0.05). Scanning electron microscope observation showed that a large amount of Staphylococcus epidermis and mixed biofilm structure were observed in the positive group, and a very small amount of bacteria was observed in the negative group with no mixed biofilm structure. HE staining of surrounding tissue showed inflammatory cell infiltration in both groups, and neutrophils and lymphocytes were more in the positive group than in the negative group. There was no significant difference in the number of bacterial infections in heart and liver between the two groups (P>0.05). The number of bacterial infections in lung and kidney in the positive group was higher than that in negative group (P<0.05).Conclusion In the mixed infection of Staphylococcus epidermidis and Candida albicans, the ica operon may strengthen the structure of the biofilm and the spread of the biofilm in vivo, leading to increased inflammatory factors, and the bacteria are difficult to remove and persist.

Citation： ZHANG Guojing, WAN Zilin, WANG Xiaoyan, HUANG Yunchao, ZHOU Youquan, CHEN Ying, LEI Yujie, QIAO Limei. In vivo study on the effects of intercellular adhesion operon of Staphylococcus epidermidis on the inflammation associated with bacteria-fungal mixed biofilm. Chinese Journal of Reparative and Reconstructive Surgery, 2021, 35(10): 1328-1335. doi: 10.7507/1002-1892.202104061 Copy

1.	Dabas Y, Xess I, Kale P. Molecular and antifungal susceptibility study on trichosporonemia and emergence of Trichosporon mycotoxinivorans as a bloodstream pathogen. Med Mycol, 2017, 55(5): 518-527.
2.	Jung P, Mischo CE, Gunaratnam G, et al. Candida albicans adhesion to central venous catheters: Impact of blood plasma-driven germ tube formation and pathogen-derived adhesins. Virulence, 2020, 11(1): 1453-1465.
3.	Tsui C, Kong EF, Jabra-Rizk MA. Pathogenesis of Candida albicans biofilm. Pathog Dis, 2016, 74(4): ftw018. doi: 10.1093/femspd/ftw018.
4.	Holt JE, Houston A, Adams C, et al. Role of extracellular polymeric substances in polymicrobial biofilm infections of Staphylococcus epidermidis and Candida albicans modelled in the nematode Caenorhabditis elegans. Pathog Dis, 2017, 75(5): ftx052. doi: 10.1093/femspd/ftx052.
5.	Schilcher K, Horswill AR. Staphylococcal biofilm development: structure, regulation, and treatment strategies. Microbiol Mol Biol Rev, 2020, 84(3): e00026-19. doi: 10.1128/MMBR.00026-19.
6.	Valle J, Solano C, García B, et al. Biofilm switch and immune response determinants at early stages of infection. Trends Microbiol, 2013, 21(8): 364-371.
7.	陳雅, 葉聯華, 黃云超. 葡萄球菌生物膜形成影響因素的研究進展. 中國醫藥導報, 2017, 14(36): 37-42.
8.	陳鵬, 李波, 彭智, 等. ica操縱子對表皮葡萄球菌在不同骨科植入物表面黏附和生物膜形成的影響. 重慶醫學, 2018, 47(17): 2275-2278, 2284.
9.	葉聯華, 黃云超, 楊達寬, 等. 表皮葡萄球菌ica操縱子與聚氯乙烯材料表面細菌生物膜形成的關系. 中華醫院感染學雜志, 2010, 20(24): 3841-3843.
10.	Mirzaei B, Faridifar P, Shahmoradi M, et al. Genotypic and phenotypic analysis of biofilm formation Staphylococcus epidermidis isolates from clinical specimens. BMC Res Notes, 2020, 13(1): 114. doi: 10.1186/s13104-020-04965-y.
11.	Carolus H, Van Dyck K, Van Dijck P. Candida albicans and Staphylococcus species: A threatening twosome. Front Microbiol, 2019, 10: 2162. doi: 10.3389/fmicb.2019.02162.
12.	申友亮, 朱同娥, 張靖靖, 等. 構建膝關節假體感染兔模型: 體內環境因素對表皮葡萄球菌及生物膜的影響. 中國組織工程研究, 2015, 19(39): 6240-6245.
13.	徐曉耘, 袁詠梅, 陳曉君, 等. 呼吸機相關性肺炎患者血清IL-8, TNF-α, MCP-1的變化. 中華醫院感染學雜志, 2020, 30(17): 2624-2627.
14.	黃云超, 張爾永, 石應康, 等. 兔體內細菌對人工心臟瓣膜的粘附及清除研究. 生物醫學工程與臨床, 2004, 8(2): 61-64.
15.	黃筱雪, 李燕. 白假絲酵母生物膜研究進展. 中華醫院感染學雜志, 2019, 29(21): 3350-3354.
16.	隋明亮, 吳長江, 黃超發, 等. 降鈣素原和白介素6對危重患者導管相關性血行感染的早期診斷及預后價值. 上海交通大學學報 (醫學版), 2013, 33(11): 1491-1495.
17.	Dai L, Yang L, Parsons C, et al. Staphylococcus epidermidis recovered from indwelling catheters exhibit enhanced biofilm dispersal and “self-renewal” through downregulation of agr. BMC Microbiol, 2012, 12: 102. doi: 10.1186/1471-2180-12-102.
18.	Pammi M, Liang R, Hicks J, et al. Biofilm extracellular DNA enhances mixed species biofilms of Staphylococcus epidermidis and Candida albicans. BMC Microbiol, 2013, 13: 257. doi: 10.1186/1471-2180-13-257.
19.	Vila T, Kong EF, Montelongo-Jauregui D, et al. Therapeutic implications of C.albicans-S.aureus mixed biofilm in a murine subcutaneous catheter model of polymicrobial infection. Virulence, 2021, 12(1): 835-851.
20.	Nogueira F, Sharghi S, Kuchler K, et al. Pathogenetic impact of bacterial-fungal interactions. Microorganisms, 2019, 7(10): 459. doi: 10.3390/microorganisms7100459.
21.	Pichard DC, Freeman AF, Cowen EW. Primary immunodeficiency update: Part Ⅱ. Syndromes associated with mucocutaneous candidiasis and noninfectious cutaneous manifestations. J Am Acad Dermatol, 2015, 73(3): 367-381.
22.	d’Enfert C, Kaune AK, Alaban LR, et al. The impact of the fungus-host-microbiota interplay upon Candida albicans infections: current knowledge and new perspectives. FEMS Microbiol Rev, 2021, 45(3): fuaa060. doi: 10.1093/femsre/fuaa060.
23.	阮文鵬. 表皮葡萄球菌agrC特異結合多肽對大鼠中心靜脈插管感染作用研究. 昆明: 昆明醫科大學, 2018.
24.	Bergeron AC, Seman BG, Hammond JH, et al. Candida albicans and Pseudomonas aeruginosa interact to enhance virulence of mucosal infection in transparent zebrafish. Infect Immun, 2017, 85(11): e00475-417. doi: 10.1128/IAI.00475-17.
25.	Lianhua Y, Yunchao H, Guangqiang Z, et al. The effect of iatrogenic Staphylococcus epidermidis intercellar adhesion operon on the formation of bacterial biofilm on polyvinyl chloride surfaces. Surg Infect (Larchmt), 2014, 15(6): 768-773.
26.	Cue D, Lei MG, Lee CY. Genetic regulation of the intercellular adhesion locus in staphylococci. Front Cell Infect Microbiol, 2012, 2: 38. doi: 10.3389/fcimb.2012.00038.
27.	Nguyen HTT, Nguyen TH, Otto M. The staphylococcal exopolysaccharide PIA—Biosynthesis and role in biofilm formation, colonization, and infection. Comput Struct Biotechnol J, 2020, 18: 3324-3334.
28.	Rowe SE, Campbell C, Lowry C, et al. AraC-type regulator Rbf controls the Staphylococcus epidermidis biofilm phenotype by negatively regulating the icaADBC repressor SarR. J Bacteriol, 2016, 198(21): 2914-2924.
29.	覃鳳蘭. 呼吸機相關性肺炎的微生物學與炎癥指標的分析. 南寧: 廣西醫科大學, 2019.
30.	Cherifi S, Byl B, Deplano A, et al. Comparative epidemiology of Staphylococcus epidermidis isolates from patients with catheter-related bacteremia and from healthy volunteers. J Clin Microbiol, 2013, 51(5): 1541-1547.

1. Dabas Y, Xess I, Kale P. Molecular and antifungal susceptibility study on trichosporonemia and emergence of Trichosporon mycotoxinivorans as a bloodstream pathogen. Med Mycol, 2017, 55(5): 518-527.
2. Jung P, Mischo CE, Gunaratnam G, et al. Candida albicans adhesion to central venous catheters: Impact of blood plasma-driven germ tube formation and pathogen-derived adhesins. Virulence, 2020, 11(1): 1453-1465.
3. Tsui C, Kong EF, Jabra-Rizk MA. Pathogenesis of Candida albicans biofilm. Pathog Dis, 2016, 74(4): ftw018. doi: 10.1093/femspd/ftw018.
4. Holt JE, Houston A, Adams C, et al. Role of extracellular polymeric substances in polymicrobial biofilm infections of Staphylococcus epidermidis and Candida albicans modelled in the nematode Caenorhabditis elegans. Pathog Dis, 2017, 75(5): ftx052. doi: 10.1093/femspd/ftx052.
5. Schilcher K, Horswill AR. Staphylococcal biofilm development: structure, regulation, and treatment strategies. Microbiol Mol Biol Rev, 2020, 84(3): e00026-19. doi: 10.1128/MMBR.00026-19.
6. Valle J, Solano C, García B, et al. Biofilm switch and immune response determinants at early stages of infection. Trends Microbiol, 2013, 21(8): 364-371.
7. 陳雅, 葉聯華, 黃云超. 葡萄球菌生物膜形成影響因素的研究進展. 中國醫藥導報, 2017, 14(36): 37-42.
8. 陳鵬, 李波, 彭智, 等. ica操縱子對表皮葡萄球菌在不同骨科植入物表面黏附和生物膜形成的影響. 重慶醫學, 2018, 47(17): 2275-2278, 2284.
9. 葉聯華, 黃云超, 楊達寬, 等. 表皮葡萄球菌ica操縱子與聚氯乙烯材料表面細菌生物膜形成的關系. 中華醫院感染學雜志, 2010, 20(24): 3841-3843.
10. Mirzaei B, Faridifar P, Shahmoradi M, et al. Genotypic and phenotypic analysis of biofilm formation Staphylococcus epidermidis isolates from clinical specimens. BMC Res Notes, 2020, 13(1): 114. doi: 10.1186/s13104-020-04965-y.
11. Carolus H, Van Dyck K, Van Dijck P. Candida albicans and Staphylococcus species: A threatening twosome. Front Microbiol, 2019, 10: 2162. doi: 10.3389/fmicb.2019.02162.
12. 申友亮, 朱同娥, 張靖靖, 等. 構建膝關節假體感染兔模型: 體內環境因素對表皮葡萄球菌及生物膜的影響. 中國組織工程研究, 2015, 19(39): 6240-6245.
13. 徐曉耘, 袁詠梅, 陳曉君, 等. 呼吸機相關性肺炎患者血清IL-8, TNF-α, MCP-1的變化. 中華醫院感染學雜志, 2020, 30(17): 2624-2627.
14. 黃云超, 張爾永, 石應康, 等. 兔體內細菌對人工心臟瓣膜的粘附及清除研究. 生物醫學工程與臨床, 2004, 8(2): 61-64.
15. 黃筱雪, 李燕. 白假絲酵母生物膜研究進展. 中華醫院感染學雜志, 2019, 29(21): 3350-3354.
16. 隋明亮, 吳長江, 黃超發, 等. 降鈣素原和白介素6對危重患者導管相關性血行感染的早期診斷及預后價值. 上海交通大學學報 (醫學版), 2013, 33(11): 1491-1495.
17. Dai L, Yang L, Parsons C, et al. Staphylococcus epidermidis recovered from indwelling catheters exhibit enhanced biofilm dispersal and “self-renewal” through downregulation of agr. BMC Microbiol, 2012, 12: 102. doi: 10.1186/1471-2180-12-102.
18. Pammi M, Liang R, Hicks J, et al. Biofilm extracellular DNA enhances mixed species biofilms of Staphylococcus epidermidis and Candida albicans. BMC Microbiol, 2013, 13: 257. doi: 10.1186/1471-2180-13-257.
19. Vila T, Kong EF, Montelongo-Jauregui D, et al. Therapeutic implications of C.albicans-S.aureus mixed biofilm in a murine subcutaneous catheter model of polymicrobial infection. Virulence, 2021, 12(1): 835-851.
20. Nogueira F, Sharghi S, Kuchler K, et al. Pathogenetic impact of bacterial-fungal interactions. Microorganisms, 2019, 7(10): 459. doi: 10.3390/microorganisms7100459.
21. Pichard DC, Freeman AF, Cowen EW. Primary immunodeficiency update: Part Ⅱ. Syndromes associated with mucocutaneous candidiasis and noninfectious cutaneous manifestations. J Am Acad Dermatol, 2015, 73(3): 367-381.
22. d’Enfert C, Kaune AK, Alaban LR, et al. The impact of the fungus-host-microbiota interplay upon Candida albicans infections: current knowledge and new perspectives. FEMS Microbiol Rev, 2021, 45(3): fuaa060. doi: 10.1093/femsre/fuaa060.
23. 阮文鵬. 表皮葡萄球菌agrC特異結合多肽對大鼠中心靜脈插管感染作用研究. 昆明: 昆明醫科大學, 2018.
24. Bergeron AC, Seman BG, Hammond JH, et al. Candida albicans and Pseudomonas aeruginosa interact to enhance virulence of mucosal infection in transparent zebrafish. Infect Immun, 2017, 85(11): e00475-417. doi: 10.1128/IAI.00475-17.
25. Lianhua Y, Yunchao H, Guangqiang Z, et al. The effect of iatrogenic Staphylococcus epidermidis intercellar adhesion operon on the formation of bacterial biofilm on polyvinyl chloride surfaces. Surg Infect (Larchmt), 2014, 15(6): 768-773.
26. Cue D, Lei MG, Lee CY. Genetic regulation of the intercellular adhesion locus in staphylococci. Front Cell Infect Microbiol, 2012, 2: 38. doi: 10.3389/fcimb.2012.00038.
27. Nguyen HTT, Nguyen TH, Otto M. The staphylococcal exopolysaccharide PIA—Biosynthesis and role in biofilm formation, colonization, and infection. Comput Struct Biotechnol J, 2020, 18: 3324-3334.
28. Rowe SE, Campbell C, Lowry C, et al. AraC-type regulator Rbf controls the Staphylococcus epidermidis biofilm phenotype by negatively regulating the icaADBC repressor SarR. J Bacteriol, 2016, 198(21): 2914-2924.
29. 覃鳳蘭. 呼吸機相關性肺炎的微生物學與炎癥指標的分析. 南寧: 廣西醫科大學, 2019.
30. Cherifi S, Byl B, Deplano A, et al. Comparative epidemiology of Staphylococcus epidermidis isolates from patients with catheter-related bacteremia and from healthy volunteers. J Clin Microbiol, 2013, 51(5): 1541-1547.

Chinese Journal of Reparative and Reconstructive Surgery

In vivo study on the effects of intercellular adhesion operon of Staphylococcus epidermidis on the inflammation associated with bacteria-fungal mixed biofilm

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content