背景:各种纤维骨损伤的模糊和相似的组织病理学特征使得确定诊断成为挑战。需要额外的分子和组织化学工具来支持和区分这些病变,以建立具体的诊断。
方法:回顾性分析福尔马林固定石蜡包埋切片活检病灶(纤维发育不良各10例,骨化性纤维瘤,和骨水泥骨发育不良)从档案中检索到的骨钙蛋白的免疫表达(骨细胞的定量分析),使用Azan,Picrosirus,和甲苯胺蓝染色,用于评估胶原纤维的强度和定位,和血管的形态测量分析(用于评估平均血管密度为平方微米)。
结果:骨钙蛋白免疫染色阳性提示在纤维发育不良中间接发现GNAS-1基因突变,因为它是骨形成的负调节剂。在纤维骨病变中定量测量骨钙蛋白免疫阳性,纤维发育不良为14.47±3.628,而骨化纤维瘤为5.23±1.33,其次是骨水泥发育不良为2.30±1.409。甲苯胺蓝提示骨化性纤维瘤和骨水泥骨发育不良中存在奥氧塔兰纤维(抗酸水解),指向病变的发病机制。Azan染色和Picrosirus(在偏光显微镜下)有助于区分硬组织特征(70%的纤维发育不良病例仅显示洋红色成分,然后在20%的病例中混合洋红色与蓝色成分,只有10%的病例显示洋红色与蓝色边缘,而对于骨化性纤维瘤,40%的病例描绘了品红色与蓝色边界,另外40%的病例描绘了品红色与蓝色成分的混合)。在纤维发育不良中,平均血管密度也最高,为7.90±1.079(单位:Sq。微米面积),其次是骨化纤维瘤和骨水泥发育不良。
结论:单独使用苏木精和曙红对纤维骨病变的诊断令人困惑,因此应该得到相对简单的组织形态计量学分析的支持,以获得更好的治疗结果。在纤维骨病变的诊断阶段,病变内血管大小的评估,可靠的分子标记,和组织化学性质可以帮助区分纤维发育不良与中央骨化纤维瘤和骨水泥-骨发育不良,其他临床,影像学和病理学标准。这些参数有助于纤维骨病变的诊断决策。
BACKGROUND: Indistinct and analogous histopathological features of various fibro-osseous lesions make establishing a definitive diagnosis a challenge. There is a need for additional molecular and histochemical tools to support and differentiate these lesions in order to establish a concrete diagnosis.
METHODS: A retrospective analysis of biopsied lesions in formalin-fixed paraffin-embedded sections (10 cases each of fibrous dysplasia, ossifying fibroma, and cement-osseous dysplasia) retrieved from the archives was studied for immunoexpression of osteocalcin (quantitative analysis in osteocytes), collagen characterization using Azan, Picrosirus, and Toluidine blue stain for evaluating intensity and localization of collagen fibers, and morphometric analysis of vasculature (for evaluating mean vessel density as square microns).
RESULTS: Positive immunostaining of osteocalcin suggested mutations of the GNAS-1 gene found in fibrous dysplasia indirectly, as it is a negative regulator of bone formation. Osteocalcin immunopositivity was quantitatively measured in the fibro-osseous lesions, with fibrous dysplasia measuring 14.47 ± 3.628 as compared to ossifying fibroma measuring 5.23 ± 1.33, followed by cemento-osseous dysplasia measuring 2.30 ± 1.409. Toluidine blue suggests the presence of oxytalan fibers (resistant to acid hydrolysis) in ossifying fibroma and cemento-osseous dysplasia, pointing toward the pathogenesis of the lesion. Azan stain and Picrosirus (under a polarizing microscope) helped in distinguishing hard tissue characteristics (70% of cases of fibrous dysplasia showed only a magenta component followed by intermixed magenta with a blue component in 20% of cases and only 10% of cases showed magenta with blue borders whereas for ossifying fibroma, 40% of cases depicted magenta with blue borders along with the other 40% with intermixed magenta with blue component). The mean vessel density was also highest in fibrous dysplasia measuring 7.90 ± 1.079 (in Sq. micron area), followed by ossifying fibroma and cemento-osseous dysplasia.
CONCLUSIONS: The diagnosis of fibro-osseous lesions by hematoxylin and eosin alone is confusing and thus should be supported by relatively simple histomorphometric analysis for better treatment outcomes. At the diagnostic stage of fibro-osseous lesions, evaluation of intralesional vessel size, reliable molecular marker, and histochemical nature can aid in differentiating fibrous dysplasia from central ossifying fibroma and cemento-osseous dysplasia alongside, other clinical, radiographic and pathological criteria. These parameters help in the diagnostic decision-making of fibro-osseous lesions.