Pierrat Sébastien, Brochard-Wyart Françoise, Nassoy Pierre
Laboratoire de Physico-Chimie Curie, Unité Mixte de Recherche 168, Centre National de la Recherche Scientifique, Institut Curie, Paris, France.
Biophys J. 2004 Oct;87(4):2855-69. doi: 10.1529/biophysj.104.043695.
We investigated the mechanical strength of adhesion and the dynamics of unbinding of red blood cells to solid surfaces. Two different situations were tested: 1), native red blood cells nonspecifically adhered to glass surfaces coated with positively charged polymers and 2), biotinylated red blood cells specifically adhered to glass surfaces decorated with streptavidin, which has a high binding affinity for biotin. We used micropipette manipulation for forming and subsequently breaking the adhesive contact through a stepwise micromechanical procedure. Analysis of cell deformations provided the relation between force and contact radius, which was found to be in good agreement with theoretical predictions. We further demonstrated that the separation energy could be precisely derived from the measure of rupture forces and the cell shape. Finally, the dynamics of detachment was analyzed as a function of the applied force and the initial size of the adhesive patch. Our experiments were supported by original theoretical predictions, which allowed us to correlate the measured separation times with the molecular parameters (e.g., activation barrier, receptor-ligand characteristic length) derived from force measurements at the single bond level.
我们研究了红细胞与固体表面的黏附机械强度以及解离动力学。测试了两种不同情况:1)天然红细胞非特异性黏附于涂有带正电聚合物的玻璃表面;2)生物素化红细胞特异性黏附于装饰有链霉亲和素的玻璃表面,链霉亲和素对生物素有高结合亲和力。我们使用微量移液器操作,通过逐步微机械程序形成并随后打破黏附接触。对细胞变形的分析给出了力与接触半径之间的关系,发现该关系与理论预测高度吻合。我们进一步证明,分离能可从破裂力的测量和细胞形状精确推导得出。最后,分析了解离动力学作为所施加力和黏附斑初始大小的函数。我们的实验得到了原始理论预测的支持,这使我们能够将测量的分离时间与从单键水平力测量得出的分子参数(例如,活化能垒、受体 - 配体特征长度)相关联。