Zhang Jun, Peng Ching-An
Department of Biological Engineering, University of Idaho, Moscow, Idaho, USA.
J Biomed Mater Res A. 2021 Mar;109(3):365-373. doi: 10.1002/jbm.a.37029. Epub 2020 Jun 26.
CD200 is an anti-inflammatory transmembrane glycoprotein in the immunoglobulin superfamily. The interaction of CD200 and its receptor CD200R has shown to inhibit inflammatory response of myeloid cells to foreign materials. The purpose of this study is to create a CD200 immobilized biomaterial surface through polydopamine coating to suppress macrophage cell adhesion and reduce inflammatory cytokine secretion accordingly by macrophages. In this study, tissue-culture treated polystyrene (TCPS) surface was modified with biotin through polydopamine coating. Purified CD200-streptavidin fusion protein was then immobilized onto the biotinylated TCPS surface through the high affinity between biotin and streptavidin. Mouse J774A.1 macrophages were seeded on CD200-immobilized TCPS surface to evaluate the effect of CD200 on preventing macrophage attachment. The effects of CD200-immobilized TCPS surface on pro-inflammatory cytokine secretion from J774A.1 macrophages were measured by enzyme-linked immunosorbent assay. As a result, CD200-immobilized TCPS surface suppressed macrophage attachment for up to 9 hr. The level of IL-6 and TNF-α secreted from J774A.1 macrophages interacted with CD200-coated TCPS surface was reduced by 36.3% and 32.4%, respectively.
CD200是免疫球蛋白超家族中的一种抗炎跨膜糖蛋白。CD200与其受体CD200R的相互作用已显示可抑制髓样细胞对外来物质的炎症反应。本研究的目的是通过聚多巴胺涂层创建一种固定有CD200的生物材料表面，以抑制巨噬细胞黏附，并相应减少巨噬细胞分泌炎性细胞因子。在本研究中，通过聚多巴胺涂层用生物素修饰组织培养处理过的聚苯乙烯（TCPS）表面。然后，通过生物素与链霉亲和素之间的高亲和力，将纯化的CD200-链霉亲和素融合蛋白固定在生物素化的TCPS表面。将小鼠J774A.1巨噬细胞接种在固定有CD200的TCPS表面，以评估CD200对防止巨噬细胞附着的作用。通过酶联免疫吸附测定法测量固定有CD200的TCPS表面对J774A.1巨噬细胞分泌促炎细胞因子的影响。结果，固定有CD200的TCPS表面在长达9小时内抑制了巨噬细胞附着。与包被有CD200的TCPS表面相互作用的J774A.1巨噬细胞分泌的IL-6和TNF-α水平分别降低了36.3%和32.4%。
