强直性脊柱炎棘上棘间韧带成纤维细胞在体外培养中的成骨作用

OSTEOGENIC ROLE PLAYED BY FIBROBLASTS OF THE SUPRA INTERSPINOUS LIGAMENT OF ANKYLOSING SPONDYLITIS IN VITRO

目的：观察强直性脊柱炎（ＡＳ）棘上棘间韧带成纤维细胞在体外培养条件下，是否具有成骨能力。方法：采用组织块种植法，培养ＡＳ晚期棘上韧带成纤维细胞，对培养的ＡＳ成纤维细胞进行显微观察和组化染色。并用正常人的同名细胞作对照培养。结果：传代培养后的成纤维细胞分泌不透光结节，结节融合扩大，数量增多，散布于成纤维细胞之间。培养细胞经ＡＲＳ钙盐染色，甲苯胺蓝胶原染色，均呈阳性反应，表明含有钙盐及胶原成分。培养早期成纤维细胞经ＡｌｃａｉｎＢｌｕｓ染色胞浆即呈阳性反应，说明ＡＳ韧带成纤维细胞培养早期能合成分泌粘多糖。经四环素活体标记，细胞分泌的结节在蓝紫色荧光显微镜下显示金黄色荧光，证明是新生骨组织。此外还发现细胞内和培养液中均有碱性磷酸酶阳性颗粒。而对照组细胞与此有明显的区别。结论：ＡＳ韧带成纤维细胞具有强烈的成骨能力，它在ＡＳ脊柱韧带发展为“竹节椎”

Objective:To observe if the fibroblasts from AS supra interspinous ligament have osteogenic ability in vitro .Methods:Fibrous tissues were collected from the AS supra-interspinous ligament and cultured in vitro .As a control group,normal specimen was collected from a resection of nucleus pulposus of lumbal.The cultured fibroblasts were observed by contrast microscope and histochemical study.Results:In AS group,many small nodules were secreted by subcultured fibroblasts.They can mix together,multiply and distribute among the fibroblasts.Positive reaction was found from the cells and nodules,which were stained by alizarin red S and toludine blue.It showed that the cells and the nodules contained calcium and collagen.The cytoplasm of the cells revealed positive reaction to alcain blue stain in the early stage of the subculture,indicating that the fibroblasts could synthesize and secrete acid mucopolysaccharide,The nodules revealed golden yellow fluorescence when using tetracyline label,denoting newly formed bone.In addition,AKP positive grains were found within the cells and in the culture medium.There was significant difference between the 2 groups.The control group only showed positive reactions weakly in some stains.Conclusion;This experiment shows that AS fibroblasts have osteogenic ability in vitro .The cells might play an important role in the course of AS spine ligament developing “bamboo spine”.