强直性脊柱炎棘上棘间韧带的扫描电镜与成纤维细胞培养研究

Scanning electron microscopy of supra interspinous ligament in cases of ankylosing spondilitis and culture of fibroblasts in vitro

目的 观察强直性脊柱炎 (AS)棘上棘间韧带的超微结构和探讨成纤维细胞在韧带骨化中的作用 ,找出AS韧带中胶原紊乱排列和钙颗粒沉积是否由成纤维细胞活动引起。方法 采用扫描电镜—X线能谱仪联机对AS脊柱韧带进行观察。采用细胞培养法对AS棘上棘间韧带成纤维细胞进行体外培养 ,显微观察和组化染色等。结果 扫描电镜显示韧带上的胶原排列紊乱 ,其间有钙颗粒沉积 ,X线能谱能显示出钙峰。细胞培养观察到AS成纤维细胞增殖活跃 ,分泌颗粒旺盛。细胞经AlcainBlue、甲苯胺蓝、ARS染色均呈阳性反应。AKP酶测试亦显示有多量AKP存在于胞体内外。结论 AS的成纤维细胞能分泌各种成骨基质。

Objective To observe the ultrastructure of supra interspinous ligament of ankylosing spondilitis(AS),to evaluate the role of fibroblasts in ossification of ligament,and to discuss if disorderly arrangement of collagen and sedimentation of calcium granule in AS are induced by fibroblasts.Methods Scanning electron microscope (SME) and X ray microanalytic system were used to examine spinal ligament of AS.The fibroblasts of supra interspinous ligament of AS were cultured in vitro and the cells were observed by microscope after histochemical staning.Results The results of SME examination displayed that the collagen fibers of ligament arranged in disorder and calcium granule deposited in gaps of the collagen of AS which can be seen as a calcium peak by X ray microanalytic system examination.The results of cell culture showed that fibroblasts which secreted a lot of granules were in active proliferation conditions.The fibroblasts showed possitive reaction when they were stained by Alcain Blue,Toluidin Blue and Alizarin Red S.The results of AKP enzyme test showed that there were many AKP within and outside the fibroblast cells.Conclusion The fibroblasts of AS can secrete different kinds of osteogenic matrix which is closely related to disordered arrangement of collagen and sedimentation of calcium granule of spinal ligament of AS.