导电性聚乙烯醇复合水凝胶对巨噬细胞和血管内皮细胞的作用研究  

Effects of a Carbon Fiber Reinforced Polyvinyl Alcohol Conductive Hydrogel on Macrophages and Vascular Endothelial Cells

作　　者：校搏[1] 许仕琳 吴凤新[2] 刘暄欣 黄宇光[1] 许海燕[2] Xiao Bo;Xu Shilin;Wu Fengxin;Liu Xuanxin;Huang Yuguang;Xu Haiyan(Department of Anesthesiology,Peking Union Medical College Hospital,Chinese Academy of Medical Science&Peking Union Medical College,Beijing 100730,China;Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100005,China)

机构地区：[1]中国医学科学院北京协和医学院,北京协和医院麻醉科,北京100730 [2]中国医学科学院基础医学研究所北京协和医学院基础学院,北京100005

出　　处：《中国生物医学工程学报》2022年第6期717-723,共7页Chinese Journal of Biomedical Engineering

基　　金：国家自然科学基金(81971366);中国医学科学院医学与健康科技创新工程(CIFMS 2021-I2M-1-006)。

摘　　要：水凝胶敷料对免疫细胞和血管内皮细胞的作用与创伤的愈合密切相关。本课题研究了碳纤维/聚乙烯醇复合水凝胶(PVA/CFs)对小鼠巨噬细胞的作用以及对原代人脐静脉内皮细胞间粘附连接的作用。在PVA材料及CF含量为1.5%、3%、5%的PVA/CFs材料上分别培养以上两种细胞,使用实时PCR方法和ELISA试剂盒检测小鼠巨噬细胞(RAW264.7)促炎因子TNF-α、IL-1β和IL-6在基因和蛋白水平上的表达;通过CCK-8试剂盒检测RAW264.7和原代人脐静脉内皮细胞(HUVEC)在PVA/CFs上的存活与增殖;采用Western blot和激光共聚焦显微镜检测并观察HUVEC细胞在PVA/CFs上的血管内皮钙黏蛋白(VE-cadherin)表达;所有定量检测实验中至少设3个重复样(n≥3)。与PVA相比,PVA/CFs复合水凝胶未显著改变RAW264.7细胞TNF-α、IL-1β和IL-6的基因表达及相应蛋白的分泌水平,各组间无显著性差异(P>0.05),而阳性对照样品脂多糖(LPS)使巨噬细胞的TNF-α、IL-1β和IL-6分泌量显著升高,分别是其对照组的2.35、4.27和27.1倍。在PVA/CFs上生长的HUVEC细胞的VE-cadherin的表达增加,提示PVA/CFs具有促进细胞间粘附连接的作用。PVA/CFs可支持上述两种细胞的正常生长与增殖,在培养24、48、72 h后,各PVA/CFs组的细胞活性与PVA组相比无显著性差异(P>0.05)。复合水凝胶PVA/CFs能够支持巨噬细胞和内皮细胞的正常生长,未引起巨噬细胞的炎症反应,并且可促进内皮细胞间形成粘附连接。The effects of hydrogel dressings on the immune cells and vascular endothelial cells are closely related to wound healing.This work is aimed to investigate the effects of a carbon fiber reinforced polyvinyl alcohol conductive hydrogel(PVA/CFs)on inflammatory response in mouse macrophages and the formation of adherent junctions between HUVEC cells.The above two kinds of cells were cultured on PVA and PVA/CFs with CF content of 1.5%,3%and 5%,respectively.The expression of TNF-α,IL-1βand IL-6 in inflammatory response genes of macrophages(RAW264.7)was detected by real-time PCR.The cytokins secretion of TNF-α,IL-1βand IL-6 were tested by ELISA kits.The growth and proliferation of human umbilical vein endothelial cells(HUVEC)and RAW264.7 on PVA/CFs were detected by CCK-8 assay.The protein expression of VE-Cadherin in HUVEC was determined by laser scanning confocal microscopy and western blot assay.All quantitative detection experiments were set at least three replicates(n≥3).PVA/CFs did not significantly change the gene expression levels of TNF-α,IL-1βand IL-6 and the cytokins secretion levels in RAW264.7 cells compared with PVA.All groups were no significant difference(P>0.05).However,the secretion of TNF-α,IL-1βand IL-6 in RAW264.7 cells treated with LPS was increased by 2.35,4.27 and 27.1 folds of that in the control group,respectively.The expression of VE-cadherin in HUVEC cells on PVA/CFs was increased,suggesting that PVA/CFs could strengthen adherent junctions of endothelial cells.In addition,PVA/CFs could support the growth and proliferation of RAW264.7 and HUVEC cells,and there was no significant difference in cell viability between PVA/CFs and PVA groups(P>0.05).PVA/CFs not only did not cause any severe inflammatory responses of macrophages,but also supported the growth and proliferation of macrophages and endothelial cells,and promoted the formation of integrated adherent junctions between endothelial cells.

关 键 词：碳纤维 聚乙烯醇 水凝胶 巨噬细胞 血管内皮细胞 

分 类 号：R318[医药卫生—生物医学工程]