Garbacz Katarzyna, Jarzembowski Tomasz, Kwapisz Ewa, Daca Agnieszka, Witkowski Jacek
Department of Oral Microbiology, Faculty of Medicine, Medical University of Gdańsk, Gdańsk, Poland.
Department of Medical Microbiology, Faculty of Medicine, Medical University of Gdańsk, Gdańsk, Poland.
J Oral Microbiol. 2018 Oct 23;11(1):1536193. doi: 10.1080/20002297.2018.1536193. eCollection 2019.
We used flow cytometry to compare the phagocytic activity of monocytes against strains (both biofilm and planktonic cells) isolated from denture wearers and non-wearers. Staphylococcal strains were cultured in Brain Heart Infusion broth in both planktonic and biofilm form and were stained with a fluorescent reporter (propidium iodide) and incubated with monocytes. The fluorescence of the monocytes containing phagocytized bacteria was determined by flow cytometry and normalized to that of the bacterial strains used in the experiment. Staphylococcal strains from denture wearers caused greater activation of monocytes but were less prone to phagocytosis. The percentage of monocytes containing bacterial cells after exposition to staphylococcal strains varied from 2.7% to 81.4% for planktonic cells. For biofilm-released cells, this value ranged from 0.6% to 36.2%. The effectiveness of phagocytosis, estimated based on an increase in monocyte fluorescence, amounted to 32.4 and 71 FL2 units for the biofilm and planktonic cells, respectively. The lesser efficiency of phagocytosis against biofilm in denture wearers suggests that they might have been colonized with the strains which were less prone to eradication than those from non-wearers.
我们使用流式细胞术比较了单核细胞对从戴假牙者和未戴假牙者分离出的菌株(生物膜和浮游细胞)的吞噬活性。葡萄球菌菌株以浮游和生物膜形式在脑心浸液肉汤中培养,并用荧光报告染料(碘化丙啶)染色,然后与单核细胞一起孵育。通过流式细胞术测定含有吞噬细菌的单核细胞的荧光,并将其标准化为实验中使用的细菌菌株的荧光。来自戴假牙者的葡萄球菌菌株引起单核细胞的更大激活,但更不易被吞噬。暴露于葡萄球菌菌株后,含有细菌细胞的单核细胞百分比对于浮游细胞而言从2.7%到81.4%不等。对于生物膜释放的细胞,该值范围为0.6%至36.2%。基于单核细胞荧光增加估计的吞噬效率,生物膜细胞和浮游细胞分别为32.4和71 FL2单位。对戴假牙者生物膜的吞噬效率较低表明,他们可能被那些比未戴假牙者更不易根除的菌株定植。