Figure S1 - Evidence for Regulated Interleukin-4 Expression in Chondrocyte-Scaffolds under In Vitro Inflammatory Conditions

Role of transfection and/or stimulation on the expression profile of selected markers of inflammatory arthritis. Chondrocytes in monolayer culture were treated just with transfection reagents (non-transfected) or mock transfected using empty pcDNA3.1 and pCOX-2 vectors under both stimulatory and non-stimulatory conditions. It was shown that there were only basal levels of expression of IL-1β, IL-6, iNOS and COX-2 in non-transfected, mock transfected and pcDNA.IL4 and pCOX-2-IL4 transfected cells without stimulation with rcIL-1β and rcTNFα. In contrast, on stimulation with rcIL-1β (100 ng/ml) and rcTNFα (50 ng/ml) for 96 h, only pcDNA.IL4 and pCOX-2-IL4 transfected cells were able to show a down-regulation of markers of inflammatory arthritis compared to non-transfected and mock transfected cell. This clearly indicate that the down-regulation of markers of inflammatory arthritis was exclusively due to IL-4 expression from the IL-4 containing constructs. (EPS)

转染及/或刺激对炎症性关节炎选定标志物表达谱的调控作用。本研究以单层培养的软骨细胞为实验对象，分别仅用转染试剂处理（未转染组），或在刺激与非刺激两种条件下，采用空pcDNA3.1与pCOX-2载体进行模拟转染。结果显示，在未用重组白细胞介素1β（rcIL-1β）与重组肿瘤坏死因子α（rcTNFα）刺激的情况下，未转染组、模拟转染组以及pcDNA.IL4、pCOX-2-IL4转染组的细胞中，IL-1β、IL-6、诱导型一氧化氮合酶(inducible nitric oxide synthase, iNOS)与环氧化酶2(cyclooxygenase 2, COX-2)仅呈现基础表达水平。与之相反，经浓度为100 ng/ml的rcIL-1β与50 ng/ml的rcTNFα刺激96小时后，相较于未转染组与模拟转染组细胞，仅pcDNA.IL4及pCOX-2-IL4转染组细胞可表现出炎症性关节炎相关标志物的表达下调。这明确表明，炎症性关节炎相关标志物的表达下调完全源于携带IL-4的重组载体所表达的IL-4。(EPS)