Wang Yiming, Wei Wenxin, Han Yan
Department of Plastic and Reconstructive Surgery, the First Medical Central of Chinese PLA General Hospital, Beijing, 100853, P.R.China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2020 Mar 15;34(3):373-381. doi: 10.7507/1002-1892.201904010.
To preliminary explore the effect of decellularized adipose tissue (DAT) combined with vacuum sealing drainage (VSD) on wound inflammation in pigs.
The DAT was prepared through the process of freeze-thaw, enzymatic digestion, organic solvent extraction, and vacuum freeze-drying. The appearance of DAT was observed before and after freeze-drying. HE staining was used to observe its structure and acellular effect. Eighteen male Bama minipigs were recruited, and four dorsal skin soft tissue wounds in diameter of 4 cm were made on each pig and randomly divided into 4 groups for different treatments. The wounds were treated with DAT combined with VSD in DAT/VSD group, DAT in DAT group, VSD in VSD group, and sterile gauze dressing in control group. HE staining was performed at 3, 7, 10, and 14 days after treatment. Moreover, the expressions of inflammatory factors [interleukin 1β (IL-1β), IL-6, and tumor necrosis factor α (TNF-α)], as well as the phenotypes of M1 and M2 macrophage phenotypic markers [inducible nitric oxide synthase (iNOS) and arginase 1 (ARG-1)] were detected by real-time fluorescence quantitative PCR (qRT-PCR). ELISA was used to determine the content of iNOS and ARG-1.
General observation and HE staining showed that DAT obtained in this study had a loose porous structure without cells. The neutrophils of wounds were significantly less in DAT/VSD group than in control group and DAT group ( <0.05) at 3 days after treatment, and the difference was not significant ( >0.05) between DAT/VSD group and VSD group. And the neutrophils were significantly less in DAT/VSD group than in other three groups ( <0.05) at 7, 10, and 14 days. The mRNA expressions of IL-1β, IL-6, TNF-α, and iNOS were significantly lower in DAT/VSD group than in other three groups at 3, 7, 10, and 14 days ( <0.05), while the mRNA expression of ARG-1 was significantly higher in DAT/VSD group than in other three groups ( <0.05). ELISA showed that the content of iNOS was significantly lower in DAT/VSD group than in other three groups at 3, 7, 10, and 14 days ( <0.05), while the content of ARG-1 was significantly higher in DAT/VSD group than in other three groups ( <0.05).
DAT combined with VSD can significantly reduce inflammatory cell infiltration during wound healing, regulate the expressions of inflammatory factors and macrophage phenotype, and the effect is better than single use of each and conventional dressing change.
初步探讨去细胞脂肪组织（DAT）联合封闭式负压引流（VSD）对猪伤口炎症的影响。
通过冻融、酶消化、有机溶剂萃取及真空冷冻干燥制备DAT。观察冻干前后DAT的外观。采用苏木精-伊红（HE）染色观察其结构及去细胞效果。选取18只雄性巴马小型猪，每只猪制作4个直径4 cm的背部皮肤软组织伤口，随机分为4组进行不同处理。DAT/VSD组伤口用DAT联合VSD处理，DAT组用DAT处理，VSD组用VSD处理，对照组用无菌纱布换药。处理后3、7、10和14天进行HE染色。此外，采用实时荧光定量聚合酶链反应（qRT-PCR）检测炎症因子[白细胞介素1β（IL-1β）、IL-6和肿瘤坏死因子α（TNF-α）]的表达，以及M1和M2巨噬细胞表型标志物[诱导型一氧化氮合酶（iNOS）和精氨酸酶1（ARG-1）]的表型。采用酶联免疫吸附测定（ELISA）法测定iNOS和ARG-1的含量。
大体观察和HE染色显示，本研究获得的DAT具有疏松多孔结构，无细胞。处理后3天，DAT/VSD组伤口中性粒细胞明显少于对照组和DAT组（P<0.05），DAT/VSD组与VSD组差异无统计学意义（P>0.05）。处理后7、10和14天，DAT/VSD组中性粒细胞明显少于其他三组（P<0.05）。处理后3、7、10和14天，DAT/VSD组IL-1β、IL-6、TNF-α和iNOS的mRNA表达明显低于其他三组（P<0.05），而DAT/VSD组ARG-1的mRNA表达明显高于其他三组（P<0.05）。ELISA结果显示，处理后3、7、10和14天，DAT/VSD组iNOS含量明显低于其他三组（P<0.05），而DAT/VSD组ARG-1含量明显高于其他三组（P<0.05）。
DAT联合VSD可显著减少伤口愈合过程中的炎症细胞浸润，调节炎症因子表达和巨噬细胞表型，效果优于单独使用及传统换药。
