Rose Stacey L, Babensee Julia E
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, 313 Ferst Drive, Atlanta, Georgia 30332-0535, USA.
J Biomed Mater Res A. 2008 Mar 1;84(3):661-71. doi: 10.1002/jbm.a.31305.
Model in vitro culturing systems were developed to analyze roles of biomaterial-induced leukocyte activation on endothelial cell (EC) and smooth muscle cell (SMC) phenotype, and their crosstalk. Isolated monocytes or neutrophils were pretreated with model biomaterial beads and applied directly to "more secretory" (cultured in media containing 5% fetal bovine serum) or forced contractile (serum and growth factor starved) human aortic SMCs (HASMCs), or to the human aortic EC (HAEC) surface of HAEC/HASMC cocultures (HASMC phenotype varied to be "more or less secretory") for 5 or 24 h of static culture. Surface expression of proinflammatory [ICAM-1, VCAM-1, E-selectin], procoagulant (tissue factor), and anticoagulant (thrombomodulin) markers, as well as HAEC proliferation, were assessed by flow cytometry. Incubation of HAEC with biomaterial-pretreated monocytes (and neutrophils to lesser degree) suppressed HAEC proliferation and induced a proinflammatory/procoagulant HAEC phenotype. This HAEC phenotype was amplified in coculture with "more secretory" HASMCs and subdued in coculture with "less secretory" HASMCs. Direct incubation of biomaterial-pretreated monocytes or neutrophils with "more secretory" HASMCs further increased HASMC ICAM-1 and tissue factor expression. Direct incubation of biomaterial-pretreated monocytes or neutrophils with forced contractile HASMCs upregulated ICAM-1, VCAM-1, and tissue factor expression above the presence of serum-containing media alone.
开发了体外培养系统，以分析生物材料诱导的白细胞活化对内皮细胞（EC）和平滑肌细胞（SMC）表型及其相互作用的影响。将分离的单核细胞或中性粒细胞用模型生物材料珠预处理，然后直接应用于“分泌更多”（在含有5%胎牛血清的培养基中培养）或强制收缩（血清和生长因子饥饿）的人主动脉平滑肌细胞（HASMC），或应用于HAEC/HASMC共培养物的人主动脉内皮细胞（HAEC）表面（HASMC表型变化为“或多或少分泌”），进行5或24小时的静态培养。通过流式细胞术评估促炎[细胞间黏附分子-1（ICAM-1）、血管细胞黏附分子-1（VCAM-1）、E-选择素]、促凝血（组织因子）和抗凝血（血栓调节蛋白）标志物的表面表达以及HAEC增殖。用生物材料预处理的单核细胞（和程度较轻的中性粒细胞）孵育HAEC可抑制HAEC增殖并诱导促炎/促凝血的HAEC表型。这种HAEC表型在与“分泌更多”的HASMC共培养时会放大，而在与“分泌较少”的HASMC共培养时会减弱。将生物材料预处理的单核细胞或中性粒细胞与“分泌更多”的HASMC直接孵育可进一步增加HASMC的ICAM-1和组织因子表达。将生物材料预处理的单核细胞或中性粒细胞与强制收缩的HASMC直接孵育，可使ICAM-1、VCAM-1和组织因子的表达上调，高于仅存在含血清培养基时的表达。
