胶联羊膜的组织形态、细胞相容性及抗拉力特性研究

Histomorphology,cellular biocompatibility and tensile force resistance property of fibrin-binding amniotic membrane in vitro

目的研究胶联羊膜的组织形态、体外细胞相容性及抗拉力特性,探讨该材料用于眼表移植的可行性和优越性。方法使用光镜和扫描电镜观察胶联羊膜的组织形态改变;以单层胶联羊膜为载体行角膜上皮细胞的原代培养,观察细胞生长覆盖面积;在电子万能试验机上测试胶联羊膜的拉伸性能,以单层羊膜和双层羊膜作对照,得出弹性模量。结果新制备的胶联羊膜成一整体,羊膜、纤维蛋白胶结构完整、粘合紧密,保存胶联羊膜外观完整,稍薄,但仍粘合紧密,其纤维蛋白胶面结构较模糊;体外培养7d左右胶联羊膜上即形成单层上皮细胞,第2～7天的细胞生长覆盖面积差异有统计学意义(P<0.05);胶联羊膜的弹性模量小于单、双层羊膜(P<0.01)。结论羊膜和纤维蛋白胶可结合制成胶联羊膜,该材料粘合紧密,镜下形态规则,可作为上皮细胞种植的载体,其柔韧性和可塑性优于单、双层羊膜。

Objective To investigate the histomorphology, cellular biocompatibility in vitro and tensile force resistance property of fibrin-binding amniotic membrane （FBAM） and to explore its feasibility and superiority using as ocular surface graft. Methods After observing the morphology of FBAM under light microscope and scanning electron microscope, we did primary culture of corneal epithelial cells, taking monolayer fibrinbinding amniotic membrane as carrier, then observed cell growth overlay area. The elongation nature of FBAM was tested on electronic universal testing machine by calculating elastic modulus, using bilayer amniotic membrane （BAM） and monolayer amniotic membrane （MAM） as controls. Results The fresh FBAM was integrated as a whole, with structurally complete fibrin and amniotic membrane as well as tight binding of fibrin to amniotic membrane. The reserved FBAM was structurally complete, brittler than the fresh FBAM, while amniotic membrane still bound tightly to the fibrin, but under microscope the fiber reticular formation was fairly fuzzy. Single layer of epithelial cells was formed on FBAM after about seven-day culture in vitro. Disparity of cell growth overlay area was significant （P 〈 0. 05 ） from the 2nd day to the 7th day between experimental group and control group （denuded human amniotic membrane as control）. The elastic modulus of FBAM was smaller than that of MAM and BAM （P 〈 0. 01 ）. Conclusion Amniotic membrane and fibrin can be inte FBAM. Amniotic membrane binds to fibrin tightly. The morphous of FBAM is regular. FBAM can carrier for epithelial cells. FBAM possesses better biomechanical properties than MAM and BAM.