Department of Skincare Science, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Ann Plast Surg. 2024 Nov 1;93(5):617-623. doi: 10.1097/SAP.0000000000004092. Epub 2024 Sep 4.
Detection of critical colonization is gaining importance in wound management, but its pathophysiology remains unclear. We previously clarified that a dysbiotic wound microbiota differing from skin commensal microbiota may be involved in critical colonization and that such wounds contain fewer Forkhead box protein P3 (FOXP3)-positive cells in the tissue. However, it is not clear whether FOXP3-positive cells contribute to the development of critical colonization. Here, we examined whether inhibition of FOXP3-positive cell could induce critical colonization when the commensal microbiota was present in the wounds.
Sprague-Dawley rats were administered FK506 or vehicle to inhibit differentiation into FOXP3-positive cells. Full-thickness wounds were made on the dorsal skin and inoculated with bacterial solution (dysbiosis group) or Luria-Bertani medium (commensal group). A bacterial solution was prepared by anaerobically culturing bacteria from the skin of donor rats on an artificial dermis in Luria-Bertani medium for 72 hours. Tissues were collected on day 4 postwounding for histological evaluation.
After microbiota transplantation, excessive inflammation occurred in the FK506 + commensal group. In contrast, wounds with transplanted dysbiotic microbiota showed the same level of neutrophil infiltration, regardless of FK506 administration. Furthermore, the wound area was larger in the FK506 + commensal group than in the vehicle + commensal group on day 4 postwounding ( P = 0.01). This area was also significantly larger in both the vehicle + dysbiosis ( P = 0.01) and FK506 + dysbiosis groups ( P = 0.03) than in the vehicle + commensal group.
This study has shown that dysbiosis may be at least related to developing critical colonization, and the results suggest that FOXP3-positive cells are involved in this process. Our study may contribute to establishing new interventions that prevent critical colonization by correcting wound microbiota.
在伤口管理中，对关键定植的检测变得越来越重要，但其病理生理学仍不清楚。我们之前已经阐明，与皮肤共生菌群不同的失调性伤口微生物群可能与关键定植有关，并且这些伤口的组织中含有较少的叉头框蛋白 P3（FOXP3）阳性细胞。但是，FOXP3 阳性细胞是否有助于关键定植的发展尚不清楚。在这里，当伤口中有共生菌群时，我们检查了抑制 FOXP3 阳性细胞是否会诱导关键定植。
给 Sprague-Dawley 大鼠施用 FK506 或载体以抑制分化为 FOXP3 阳性细胞。在背部皮肤制造全层伤口，并接种细菌溶液（失调组）或 Luria-Bertani 培养基（共生组）。细菌溶液是通过将供体大鼠皮肤的细菌在 Luria-Bertani 培养基中的人工真皮上进行厌氧菌培养 72 小时而制备的。在受伤后第 4 天收集组织进行组织学评估。
在微生物群移植后，FK506 + 共生组发生过度炎症。相比之下，无论是否给予 FK506，移植了失调性微生物群的伤口均显示出相同水平的中性粒细胞浸润。此外，与受伤后第 4 天的载体+共生组相比，FK506 + 共生组的伤口面积更大（P=0.01）。与载体+共生组相比，载体+失调组（P=0.01）和 FK506 + 失调组（P=0.03）的伤口面积也明显更大。
这项研究表明，失调可能至少与发展关键定植有关，并且结果表明 FOXP3 阳性细胞参与了这一过程。我们的研究可能有助于通过纠正伤口微生物群来建立预防关键定植的新干预措施。
