Bay Lene, Kragh Kasper N, Eickhardt Steffen R, Poulsen Steen S, Gjerdrum Lise Mette R, Ghathian Khaled, Calum Henrik, Ågren Magnus S, Bjarnsholt Thomas
Costerton Biofilm Center, Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark.
Endocrinology, Department of Biomedical Science, University of Copenhagen, Copenhagen, Denmark.
Adv Wound Care (New Rochelle). 2018 Apr 1;7(4):105-113. doi: 10.1089/wound.2017.0770.
The bacterial composition and distribution were evaluated in acute standardized epidermal wounds and uninjured skin by a molecular technology benchmarked to conventional culturing. This was done to reveal whether bacterial biofilm is present in acute wounds. On the buttock of 26 healthy volunteers, 28 suction blisters were made and de-roofed. Four wounds were biopsied immediately after wounding, whereas the remaining 24 wounds were treated daily with sterile deionized water and covered with a moisture-retaining dressing. On day 4 post-wounding, swabs were obtained for culturing from the wounds and adjacent skin, and the wounds including adjacent skin were excised. Tissue sections were stained with peptide nucleic acid (PNA) fluorescence hybridization (FISH) probes, counterstained by 4',6-diamidino-2-phenylindole, and evaluated by confocal laser scanning microscopy (CLSM). No bacterial aggregates were detected at day 0. At day 4, coagulase-negative staphylococci (CoNS) were the sole bacteria identified by CLSM/PNA-FISH and culturing. CoNS was isolated from 78% of the wound swabs and 48% of the skin swabs. Bacterial aggregates (5-150 μm) were detected by PNA-FISH/CLSM in the split stratum corneum and fibrin deposits at the wound edges and in the stratum corneum and the hair follicles of the adjacent skin. The bacterial aggregates were more common ( = 0.0084) and larger ( = 0.0083) at wound edges than in the adjacent skin. Bacterial aggregates can establish in all wound types and may have clinical significance in acute wounds. Bacterial aggregates were observed at the edges of acute epidermal wounds, indicating initiated establishment of a biofilm.
通过一种以传统培养为基准的分子技术,对急性标准化表皮伤口和未受伤皮肤中的细菌组成及分布进行了评估。这样做是为了揭示急性伤口中是否存在细菌生物膜。在26名健康志愿者的臀部制作了28个抽吸水疱并去除疱顶。4个伤口在受伤后立即进行活检,其余24个伤口每天用无菌去离子水治疗,并用保湿敷料覆盖。在受伤后第4天,从伤口和相邻皮肤获取拭子进行培养,并切除包括相邻皮肤的伤口。组织切片用肽核酸(PNA)荧光杂交(FISH)探针染色,用4',6-二脒基-2-苯基吲哚复染,并通过共聚焦激光扫描显微镜(CLSM)进行评估。在第0天未检测到细菌聚集体。在第4天,凝固酶阴性葡萄球菌(CoNS)是通过CLSM/PNA-FISH和培养鉴定出的唯一细菌。CoNS从78%的伤口拭子和48%的皮肤拭子中分离出来。通过PNA-FISH/CLSM在伤口边缘的角质层分裂处和纤维蛋白沉积物以及相邻皮肤的角质层和毛囊中检测到细菌聚集体(5 - 150μm)。伤口边缘的细菌聚集体比相邻皮肤更常见(P = 0.0084)且更大(P = 0.0083)。细菌聚集体可在所有伤口类型中形成,可能在急性伤口中具有临床意义。在急性表皮伤口边缘观察到细菌聚集体,表明生物膜开始形成。