The role of osteopontin and osteopontin aptamer (OPN-R3) in fibroblast activity.
The role of osteopontin and osteopontin aptamer (OPN-R3) in fibroblast activity.
复制标题
DOI：
10.1016/j.jss.2011.07.054
复制
发表时间：
2012-07
期刊：
影响因子：
--
通讯作者：
Levinson H
中科院分区：
文献类型：
--
作者：
Hunter C;Bond J;Kuo PC;Selim MA;Levinson H
瘢痕形成被认为是由持续的炎症和过度兴奋的成纤维细胞活化引起的。骨桥蛋白（OPN）是一种促进细胞活化的细胞因子。体内OPN的缺乏减少了皮肤瘢痕形成。这表明OPN参与瘢痕形成;然而，OPN如何发挥这些促瘢痕形成作用尚不清楚。RNA适体是以高亲和力结合靶蛋白的短RNA分子。适体OPN-R3（R3）阻断OPN信号传导。R3在预防皮肤纤维化中的作用尚不清楚。使用Boyden Chambers和HEMA-3染色分析成纤维细胞迁移。倒置共聚焦显微镜用于评估成纤维细胞焦点粘附长度。通过在OPN包被的96孔板上孵育荧光染色的成纤维细胞来测量粘附。使用CellTiter 96® AQueous非放射性细胞增殖测定来研究成纤维细胞的增殖活性。利用自由浮动的胶原晶格来评估成纤维细胞收缩性。人皮肤成纤维细胞对OPN的反应是显著迁移的。与对照组相比，OPN未诱导粘着斑长度显著增加。粘附研究表明，OPN增加成纤维细胞粘附。增殖试验表明，OPN增加成纤维细胞生长。OPN增加胶原网格的成纤维细胞收缩力。R3的加入显著抑制OPN诱导的活性。OPN与瘢痕相关，并通过增加人真皮成纤维细胞的细胞迁移、粘附、增殖和收缩性发挥促瘢痕形成作用。R3阻止OPN介导的活性。OPN可用于促进不愈合伤口的闭合，并且OPN特异性适体R3可用于预防纤维化。
Scarring is believed to be caused by both persistent inflammation and overexuberant fibroblast activation. Osteopontin (OPN) is a cytokine that promotes cell activation. The absence of OPN in vivo reduces dermal scarring. This suggests that OPN is involved in scar formation; however, how OPN exerts these pro-scarring effects is unknown. RNA aptamers are short RNA molecules that bind target proteins with high affinity. The aptamer OPN-R3 (R3) blocks OPN signaling. The role of R3 in preventing dermal fibrosis is unknown. Fibroblast migration was analyzed with the use of Boyden Chambers and HEMA-3 staining. Inverted confocal microscopy was used to assess fibroblast focal adhesion length. Adhesion was measured by incubating fluorescently stained fibroblasts on OPN coated 96-well plates. CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay was utilized to investigate the proliferative activity of fibroblasts. Free floating collagen lattices were utilized to assess fibroblast contractility. Human dermal fibroblasts migrated significantly in response to OPN. OPN did not induce a significant increase in focal adhesion length compared to controls. Adhesion studies demonstrated that OPN increased fibroblast adhesion. Proliferation assays indicate that OPN increased fibroblast growth. OPN increased fibroblast contractility of collagen lattices. The addition of R3 significantly inhibited OPN induced activity. OPN is associated with scar and exerts pro-scarring effects by increasing cellular migration, adhesion, proliferation, and contractility of human dermal fibroblasts. R3 prevents OPN mediated activity. OPN may be useful for promoting closure of non-healing wounds and the OPN specific aptamer, R3, may be useful for preventing fibrosis.
登录
查看更多内容
影响因子：
8.3
作者：
Matsui, Y;Jia, N;Uede, T
通讯作者：
Uede, T
DOI：
10.1165/ajrcmb.27.2.4674
发表时间：
2002-08-01
影响因子：
6.4
作者：
Kolb, M;Bonniaud, P;Gauldie, J
通讯作者：
Gauldie, J
影响因子：
5.6
作者：
Lee, Michael Y.;Ehrlich, H. Paul
通讯作者：
Ehrlich, H. Paul
影响因子：
21.3
作者：
Jun, Joon-Il;Lau, Lester F.
通讯作者：
Lau, Lester F.
影响因子：
15.9
作者：
LIAW, L;SKINNER, MP;GIACHELLI, CM
通讯作者：
GIACHELLI, CM