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I型胶原蛋白、纤连蛋白和玻连蛋白在支持机械拉伸成骨细胞黏附方面的比较。

A comparison of type I collagen, fibronectin, and vitronectin in supporting adhesion of mechanically strained osteoblasts.

作者信息

Lacouture Mario E, Schaffer Jonathan L, Klickstein Lloyd B

机构信息

Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA.

出版信息

J Bone Miner Res. 2002 Mar;17(3):481-92. doi: 10.1359/jbmr.2002.17.3.481.

Abstract

We used an adhesion assay for cells cultured under high dynamic strain to measure human osteoblast-like HOS cell adherence to immobilized type I collagen, fibronectin, and vitronectin. These conditions were designed to model the increased forces present at unstable fractures or loose joint prostheses. At a constant, low protein-coating density (1000 molecules/microm2) and 20% cyclic strain for 24 h, type I collagen, fibronectin, and vitronectin supported 24.6 +/- 2%, 16.7 +/- 3%, and 1.1 +/- 1% adherence, respectively, which paralleled the relative number of integrin-binding sites in each protein. Thus, when the number of available binding sites was limited, strain resistance was proportional to the number of integrin-ligand interactions. In contrast, at high protein-coating densities (> or = 2,500 molecules/microm2), vitronectin supported greater adherence (45.7 +/- 2%) when compared with type I collagen (37 +/- 2%) or fibronectin (34.8 +/- 2%) and directed constitutive expression of osteopontin (OPN), which suggested that there exist discrete signals on vitronectin receptor occupancy that promoted cell adherence and survival under strain. Integrin-mediated binding was necessary for resistance to strain, as evidenced by the low levels of strain resistance observed when cells were adherent in a nonintegrin-dependent manner. These findings support the utilization of at least two distinct mechanisms (i.e., tensegrity and integrin-mediated signal transduction) by HOS cells to remain adherent and viable on exposure to mechanical forces.

摘要

我们对在高动态应变下培养的细胞进行了黏附试验,以测量人成骨样HOS细胞与固定化I型胶原、纤连蛋白和玻连蛋白的黏附情况。这些条件旨在模拟不稳定骨折或松动关节假体处存在的增加的力。在恒定的低蛋白包被密度(1000分子/微米²)和20%的循环应变下作用24小时,I型胶原、纤连蛋白和玻连蛋白分别支持24.6±2%、16.7±3%和1.1±1%的黏附,这与每种蛋白质中整合素结合位点的相对数量平行。因此,当可用结合位点数量有限时,抗应变能力与整合素-配体相互作用的数量成正比。相比之下,在高蛋白包被密度(≥2500分子/微米²)下,与I型胶原(37±2%)或纤连蛋白(34.8±2%)相比,玻连蛋白支持更高的黏附(45.7±2%),并指导骨桥蛋白(OPN)的组成性表达,这表明玻连蛋白受体占据上存在离散信号,可促进细胞在应变下的黏附和存活。整合素介导的结合对抗应变是必要的,当细胞以非整合素依赖方式黏附时观察到的低抗应变水平证明了这一点。这些发现支持HOS细胞利用至少两种不同机制(即张拉整体结构和整合素介导的信号转导)在暴露于机械力时保持黏附和存活。

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