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目的观察骨痂组织中前胶原、转化生长因子－β１（ｔｒａｎｓｆｏｒｍｉｎｇｇｒｏｗｔｈｆａｃｔｏｒβ１，ＴＧＦ－β１）基因表达模式，分析ＴＧＦ－β１在骨折愈合过程中的作用，探索骨组织冰冻切片原位杂交技术。方法采用不脱钙的大鼠骨痂组织冰冻切片进行原位杂交，观察骨痂组织中前胶原和ＴＧＦ－β１基因的表达，并与先前研究作对照。结果杂交信号清晰，定位良好，特异性高。骨折第１周末，成纤维细胞的Ⅲ型前胶原基因表达占主导，Ⅰ型前胶原ｍＲＮＡ阳性成骨细胞也出现于膜内化骨区。ＴＧＦ－β１在分化、增殖的成骨细胞以及接近成熟的软骨细胞有显著表达。骨折第２周末，Ⅱ型前胶原和ＴＧＦ－β１ｍＲＮＡ在成熟的软骨细胞大量表达，而Ⅰ型前胶原ｍＲＮＡ表达也明显增加。骨折第４周末，软骨骨痂基本被骨组织替代，见散在Ⅰ型前胶原ｍＲＮＡ表达阳性的成骨细胞。同时证实共有表型表达的现象存在。结论实验结果与以往有关研究结果的吻合，提示ＴＧＦ－β１在骨折愈合过程中，尤其在细胞分化、增殖中，起重要的调节作用。同时，也说明了此方法是一种快捷、灵敏、又不失特异性的骨组织原位杂交方法。
Objective To observe the expression pattern of procollagen and transforming growth factor-β1 (TGF-β1) in callus tissue, analyze the role of TGF-β1 in the process of fracture healing, and explore the in situ hybridization of bone tissue by frozen section. Methods The frozen sections of non-decalcified rat callus tissue were used for in situ hybridization. The expression of procollagen and TGF-β1 gene in callus tissue was observed and compared with the previous study. Results hybridization signal clear, well-positioned, high specificity. At the end of the first week of fracture, type Ⅲ procollagen gene expression was dominant in fibroblasts, and type Ⅰ procollagen mRNA positive osteoblasts also appeared in the membrane-internalized bone area. TGF-β1 is significantly expressed in differentiated and proliferating osteoblasts as well as in mature chondrocytes. At the end of the second fracture, type Ⅱ procollagen and TGF-β1 mRNA were abundantly expressed in mature chondrocytes, while type Ⅰ procollagen mRNA expression was also significantly increased. At the end of the 4th week of fracture, the cartilage callus was replaced by bone tissue, see scattered osteoblasts with type Ⅰ procollagen mRNA expression. Simultaneous confirmation of the existence of phenotypic phenotypes exists. Conclusion The experimental results agree well with those of previous studies, suggesting that TGF-β1 plays an important regulatory role in the process of fracture healing, especially in cell differentiation and proliferation. At the same time, it also shows that this method is a quick, sensitive, yet specific method of bone tissue in situ hybridization.