Shelton R M, Rasmussen A C, Davies J E
Department of Anatomy, University of Birmingham Medical School, UK.
Biomaterials. 1988 Jan;9(1):24-9. doi: 10.1016/0142-9612(88)90065-8.
The in vitro migratory morphology of neonate rat calvarial osteoblasts on positively or negatively-charged polymer substrata was observed using scanning (SEM) and transmission electron microscopy (TEM). Proteins adsorbed from the culture medium onto these charged substrata were desorbed using 2% SDS and separated using polyacrylamide gel electrophoresis (PAGE). The charge sign of the substrata affected both protein adsorption and osteoblast migratory morphology. Cells flattened and adapted so closely to positively-charged substrata that the ventral cell membrane could not be distinguished at TEM. On negatively-charged substrata the ventral cell membrane was readily visible, with only focal areas of close contact with the substratum. The depth of the ventral extracellular space varied with the surface charge-carrier species on the negative substrata. Two desorbed protein fractions (MW 220 and 30 kDaltons, respectively) were correlated with osteoblast spreading on positive and negatively-charged surfaces respectively. Another protein fraction was uniquely present on PAGE profiles desorbed from negatively-charged substrata. It was concluded that the migratory morphology of osteoblasts was influenced via the intermediary of specifically adsorbed proteins.
利用扫描电子显微镜(SEM)和透射电子显微镜(TEM)观察新生大鼠颅骨成骨细胞在带正电荷或负电荷的聚合物基质上的体外迁移形态。用2%十二烷基硫酸钠(SDS)解吸从培养基吸附到这些带电基质上的蛋白质,并用聚丙烯酰胺凝胶电泳(PAGE)进行分离。基质的电荷符号影响蛋白质吸附和成骨细胞迁移形态。细胞扁平并紧密贴附于带正电荷的基质,以至于在透射电镜下无法区分腹侧细胞膜。在带负电荷的基质上,腹侧细胞膜清晰可见,仅在与基质紧密接触的局部区域可见。腹侧细胞外空间的深度随负电荷基质上表面电荷载体种类的不同而变化。两种解吸的蛋白质组分(分子量分别为220和30千道尔顿)分别与成骨细胞在带正电荷和负电荷表面的铺展相关。另一种蛋白质组分仅存在于从带负电荷基质解吸的PAGE图谱上。得出的结论是,成骨细胞的迁移形态通过特异性吸附蛋白质的介导受到影响。