原子力显微镜对不同类型胶原蛋白与细胞黏附情况的研究

The adhesion study of different types of collagen and cell by atomic force microscope

目的研究不同类型胶原蛋白与成纤维细胞及角质形成细胞的黏附情况,并用原子力显微镜观察其相互作用。方法分别用牛肌腱胶原、鼠尾胶原和标准Ⅳ型胶原包被培养板,接种相同数量的原代角质细胞,在不同时相用活细胞电子计数仪测定未黏附细胞的数量,观察不同类型胶原所未黏附的细胞数量的多少,并用原子力显微镜观察胶原蛋白与细胞的黏附情况。结果接种后5 min,三种胶原(鼠尾胶原、牛肌腱胶原、标准Ⅳ型胶原)中未贴壁的细胞数差别较大,Ⅳ型胶原中未贴壁的细胞最少;但培养至1 h,三种胶原中未贴壁的细胞数无明显差别,三者的贴壁率几乎相同;培养至第6小时三种胶原中未贴壁的细胞数仍然无明显差别,三者的贴壁率仍然基本相近。原子力显微镜可以清晰观察到胶原及胶原与细胞间黏附的微观结构。结论牛肌腱胶原和鼠尾胶原与Ⅳ型胶原对角质形成细胞的黏附作用无明显差别,原子力显微镜可以用于观察胶原及胶原与细胞间黏附的微观结构。

Objective To study the effect of different types of collagen to the adhesion of the keratinocytes, and observe the microstructure between the collagen and the cells by atomic force microscope. Methods Culture plates were covered by bovine tendon collagen, rat tail collagen and standard type Ⅳ collagen, respectively. Then the same passage of kerationcytes was seeded in one density of cells. Un-adherent cells were counted by viable count at different phases and different types of collagen. The curve of un-adherent cells at different conditions was figured, and we observed the microstrueture between the collagen and the cells by atomic forcemicroscope. Results The adhesion ability of type IV collagen to kerationcyte was higher than that of bovine tendon collagen and rat tail collagen in the first one hour, but since that, there was no significant distinction among them. Conclusion Compared with type Ⅳ collagen, collagen from bovine tendon and rat tail have similar adhesion ability to cells. The microsa＇ucture between the collagens and the ceils is observed by atomic force microscope.