皮肤成纤维细胞在体外培养中的成骨作用

osteogenesis role played by dermal flbroblasts culturedin vitro

本实验采用新西兰家兔背部中厚皮片，通过组织培养获得成纤维细胞。成纤维细胞经传代培养８天时，发现标本中有不透光的骨小结节形成。培养３７天时，发现一些小结节扩大、延伸，形成骨小梁样结构；也有骨小片状结构形成。经四环素活体标记后，这些结节在蓝紫光荧光显微镜下显示金黄色荧光，表明它们是新生骨组织。该结构的钙盐染色、胶原染色也都呈阳性，充分反映骨结构中的钙及胶原成分，以及皮肤成纤维细胞在体外培养中的成骨作用。标本ＡＳ染色后，骨形成区可见均匀、散在分布，呈卵圆形，其直径约为１０μｍ，显示苦味酸着色的斑片状物。这种斑片状物在非骨形成区则不能见到。因此，这种能被苦味酸着色的斑片状物可能是皮肤上皮细胞（或／与内皮细胞），并诱导成纤维细胞形成骨组织。

AbstractFibroblasts were harvested by in vitro cuIture ofsplit-thickness skin of New-Zeland rabbit.Someopaque bone nodmes wete formedhth。 culture flaskswhen the fibroblasts underwent subcMture for 8 days.Some of the nodules might enlarge , extend and formtrabecula-like or flake-like structures,when the fi-broHasts underwent subculture for 37 days. The abovementioned structures revealed golden-yellow fluores-cence under the blue-violet light fluoresccence micro-scope when the specimens were labeled with Tetracy-cline intravitally,denoting that they were newlyformed bone tissues, In these structures, the calciumsaIt staining and collagen staining also showed positivereactiOn, fully reflecting the calcium and collagen com-position, Osteogenesis caused by dermal fibroblast cul-tured in vitro was thus verified. There were evenly scattered Oval spots in the os-teogenic areas, after the specimens were treated withAlcina Blue-Sirius, Red stain. Their diameter wasabout 10μm , and they stained yellow color by PicricAcid,Therefore,these spots colored yellow with Pi-cric Acid might be the dermal epithelia<and/or en-dothelia), whichinduced the fibroblasts to form thebone tissues.